KR101711513B1 - Composition for improving skin - Google Patents

Abstract

The present invention relates to a composition for improving skin. The compound represented by the formula (I) or (II) or the pharmaceutically acceptable salt thereof according to the present invention promotes skin stem cell proliferation and collagen synthesis, and exhibits an effect of inhibiting elastase activity, thereby activating skin stem cells , Skin elasticity enhancement and wrinkle improvement effect, as well as antioxidative effect by eliminating free radicals, melanin synthesis is suppressed to exhibit whitening effect, so that it can be used as a pharmaceutical composition, external preparation for skin, cosmetic composition or health food.

Description

[Composition for improving skin]

The present invention relates to a composition for skin improvement such as skin wrinkle improvement, elasticity enhancement, whitening, and antioxidation.

The skin's stem cells play an important role in maintaining the health and physiological and biochemical homeostasis of the skin. Stem cells present in the skin are also abnormally functioned due to aging, which causes various problems due to the breakdown of the homeostasis of the skin. Thus, reactivation of aging stem cells will have the same effect as turning skin age backwards.

The skin consists of three layers of skin from the outside: the epidermis, the dermis and the subcutaneous fat layer. The epidermis is the thinnest of the three layers and varies in thickness depending on the region. The epidermis consists mainly of keratinocyte cells, melanocytes that produce melanin pigment, and Langerhans cells that perform immunological functions. The dermis consists of connective tissue and matrix composed mainly of collagen and elastic fibers, and it contains nerve, blood vessels, lymphatic vessels, muscles, hair follicles, sebaceous glands and sweat glands.

Embryologically, all components of human skin are known to originate from ectoderm or mesodermal lobe. Epidermis, hair follicles, sebaceous glands and glands are originated from ectoderm, and melanocytes, nerves and special sensory receptors are derived from neuroepithelial cells. Melanocytes, which are derived from neuroectodermal cells, reach the skin 8 weeks after development, and at 4 to 5 months after development, dendritic processes are developed to move the melanocytes to surrounding keratinocytes. Thus, embryonic stem cells will undergo differentiation depending on the developmental stage, and will have characteristics appropriate to their function. After becoming adult, a certain number of stem cells remain in the tissues. There are mainly two stem cells in the skin. The first is in the hair follicle. It is known to play an important role in the regeneration of the epidermis into cells before cell differentiation occurs, and plays an important role in hair regeneration and growth. The second is the basal cell layer of the epidermis, called the interfollicular epidermis, between the hair follicle and the hair follicle. The stem cells found here play an important role in maintaining skin health by administering not only the epidermis but also the fibroblasts of the dermal layer. The stem cells here are large in quantity and easy to obtain, making them widely used in the study of skin stem cells. 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 recovery of the epithelium after injury (Non-Patent Document 1). The skin epithelial stem cells are also referred to as skin stem cells. When the skin stem cells are activated, it is possible to treat skin wounds such as trauma, promote wound healing, . Integrin β1 and integrin α6 are used as indicators of dermal stem cells, and maintenance of skin stem cells necessary for epithelial morphogenesis and differentiation has been reported to be regulated by p63 protein.

Collagen, a major component of the extracellular matrix, is a major substrate protein produced by the fibroblasts of the skin. In addition, it is an important protein occupying about 30% of the total weight of the bioprotein, and has a solid triple helix structure. Collagen forms most of the organic matter in the skin, tendons, bones and teeth, especially in bone and skin (dermis). In most other body structures, it exists as a fibrous inclusion.

Collagen is a relatively weak immunogen, partly due to the blocking of the potential antigenic determinant by the helical structure of the collagen, which also causes the collagen to be resistant to proteolysis. The main functions of collagen are known to be mechanical rigidity of skin, resistance of connective tissues and binding force of tissues, support of cell adhesion, induction of cell division and differentiation (when organism grows or healing the wound) (Non-Patent Document 2) .

Such collagen is reduced by aging and photoaging due to ultraviolet irradiation, which is known to be closely related to the formation of wrinkles in the skin (Non-Patent Document 3). Collagen also plays an important role in wound healing and can accelerate the synthesis of collagen in the damaged epithelium, allowing quick, scarless wound healing.

Conventionally, products using collagen in a composition for external application for skin such as cosmetics or ointment have been marketed in order to utilize the skin moisturizing effect and wound healing effect of collagen. However, these products apply collagen itself to the skin surface, It is impossible to expect a moisturizing action or a wound healing effect, so that it can not be said that it exhibits essential skin function improvement and wound healing function.

In order to solve this problem, there has been a growing interest in collagen synthesis promoting substances, and known collagen synthesis promoting substances include vitamin C, retinoic acid, carcinostatic factors (non-patent document 4) (Patent Document 1), betulinic acid (Patent Document 2), chlorella extract (Patent Documents 3 and 4), and the like.

However, these materials have limited use due to safety problems such as irritation and reddening when applied to the skin, or the effect is insignificant and virtually no effect of skin function improvement or wound healing can be expected. Therefore, there is a desperate need to develop a novel composition for external skin application that is safer and more effective in living body than existing skin external composition compositions.

In addition, elastin fibers form crosslinks with collagen, and are skin constituents that are important for wrinkle formation that is involved in skin elasticity. The deficiency and aggregation of elastin fibers and the dramatic increase in activity of the elastase, elastase, have been found to be one of the causes of skin wrinkles. Elastase is the only enzyme capable of degrading elastin, and inhibition of it is known to be able to radically reduce wrinkles.

Recently, the study of whitening agents as cosmetics has become increasingly necessary as skin blackness is recognized as skin aging due to ultraviolet rays, along with the improvement of living standards in Asian countries, which prefer emotionally white skin. Accordingly, substances exhibiting tyrosinase inhibitory activity, such as ascorbic acid, hydroquinone, glutathione, and arbutin, have been used in cosmetics or pharmaceuticals (Patent Documents 5 and 6) 6), and most of them are ineffective or unstable due to poor formulation. In particular, a compound such as hydroquinone exhibits a strong decolorizing action, itself has a skin sensitization and can induce skin allergies and the like. It changes side effects of normal skin and causes side effects such as causing vitiligo, Its use is limited on the side.

In addition, even if the substance inhibits the activity of tyrosinase as described above, if such a substance acts on keratinocytes, which are keratinocytes, to increase the production of factors promoting melanin biosynthesis, the whitening action is consequently weakened The active inhibitor of tyrosinase is not considered to be a good whitening agent because it can cause harmful effects, and many tyrosinase inhibitors have been developed, but most of them are known to show no excellent whitening effect.

On the other hand, active oxygen introduced from the outside of the living body or generated in the living body causes many problems such as promoting aging of the living body, cancer, and the like. Therefore, the development and research of antioxidants that inhibit oxidation by active oxygen have been performed. Conventionally, there have been many efforts to obtain an anti-aging effect by adding an antioxidant such as superoxide dismutase (SOD), vitamin E derivatives, ascorbic acid and its derivatives, and flavonoid to cosmetics. However, The use thereof is limited due to problems in terms of stability in formulation.

Accordingly, it is possible to provide a method for producing a stem cell which is more safe and effective in the living body, more stable than the substance having the existing skin stem cell activation, whitening, elasticity enhancement, wrinkle improvement and antioxidant effect, whitening, elasticity enhancement, And a component having antioxidant effect are strongly desired to be developed.

Japanese Patent Application Laid-Open No. 8-231370 Japanese Patent Application Laid-Open No. 8-208424 Japanese Patent Application Laid-Open No. 9-40526 Japanese Patent Application Laid-Open No. 10-36283 Korean Patent Registration No. 10-0923141 Korean Patent Registration No. 10-0921947

 Blanpain C. and Fuchs E., Epidermal Stem Cells of the Skin, Annual Review of Cell and Developmental Biology, Vol. 22, Pages 339-373, 2006  Van der Rest et al., Structure, Molecular Biology, and Pathology of Collagen, Annals of the New York Academy of Sciences, Vol. 580, Pages 1-588, 1990  Arthur K. Balin et al., Aging and the skin, Raven Press, New York, 1989  Cardinale G. J. and Udenfriend S., Prolyl Hydroxylase, Advances in Enzymology and Related Areas of Molecular Biology, Vol. 41, Pages 245-300, 1974

Accordingly, the present inventors have found that a compound represented by the following general formula (1) or (2) promotes stem cell proliferation and collagen synthesis of skin and inhibits elastase activity, thereby activating skin stem cells, And that the present invention has been completed. Further, it has been confirmed that the compound represented by Chemical Formula (1) or Chemical Formula (2) scavenges free radicals to exhibit antioxidative and melanin synthesis inhibiting effects, thereby completing the present invention.

[Chemical Formula 1]

(2)

Accordingly, an object of the present invention is to provide a wrinkle-improving, elasticity-enhancing, whitening and antioxidative composition comprising the compound represented by the above formula (1) or (2) as an active ingredient.

Another object of the present invention is to provide a composition for stimulating stem cell activity comprising a compound represented by the following formula (1) or (2) as an active ingredient.

As means for solving the above problems,

The present invention provides a skin wrinkle improving, elasticity enhancing, whitening and antioxidant composition comprising a compound represented by the following formula (1) or (2) as an active ingredient:

[Chemical Formula 1]

(2)

As another means for solving the above problems,

The present invention provides a composition for promoting stem cell activity comprising a compound represented by the following formula (1) or (2) as an active ingredient:

[Chemical Formula 1]

(2)

The compound represented by formula (1) or (2) according to the present invention or a pharmaceutically acceptable salt thereof promotes stem cell proliferation and collagen synthesis of skin, inhibits elastase activity, activates skin stem cells, Enhancing and wrinkle-improving effect, as well as eliminating free radicals to inhibit antioxidative and melanin synthesis, thereby exhibiting a whitening effect, which can be used in pharmaceuticals, cosmetics or health foods.

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

To improve skin wrinkles, improve skin elasticity, whitening, and antioxidant properties, it is necessary to improve skin wrinkles, elasticity, whitening and antioxidant activity at low concentrations and to absorb skin penetration Is low in volatility so as to be able to stay for a sufficient time to exhibit skin wrinkle improvement, elasticity enhancement, whitening and antioxidative effect, and the active ingredient is stably maintained on the composition or on the skin and is easy to be formulated into medicines and cosmetics, It is preferable to be skin-safe. However, the components satisfying all of the above-mentioned characteristics among the known components are not common. For example, some skin wrinkle improvement, elasticity enhancement, whitening, and antioxidant components have excellent wrinkle improvement, elasticity enhancement, whitening and antioxidant activity even at low concentration in the in vitro test, It is difficult to apply. Other active ingredients have low hydrophilicity and are difficult to formulate into medicines or cosmetics. In addition, some skin wrinkle improvement, elasticity enhancement, whitening, and antioxidant ingredients may be degraded or transformed into other compounds when exposed to heat, light, or oxygen, and the effect may already be lost before application to the skin.

As can be seen in the following examples, the compound represented by the following formula (1) or (2) exhibits collagen synthesis promoting effect, elastase activity inhibiting effect, melanin formation inhibiting effect and free radical scavenging effect at low concentration, , Elasticity enhancement, whitening, antioxidant medicines, cosmetics, and health food.

Accordingly, the present invention provides a composition for improving skin wrinkles, improving skin elasticity, skin whitening, and antioxidant comprising a compound represented by the following general formula (1) or (2) as an active ingredient.

[Chemical Formula 1]

(2)

The compounds represented by the above general formulas (1) and (2) are represented by 5α-cholestan-3β, 6α, 7α, 8,15α, 16β, 26- , 15 [beta], 16 [beta], 26-octol (Formula 2).

The method for synthesizing the compound of Chemical Formula 1 or Chemical Formula 2 is not particularly limited, and may be chemically synthesized by a method known in the art, separated from natural products, or a commercially available material.

The skin wrinkle improvement, elasticity enhancement, skin whitening, and antioxidant pharmaceutical composition of the present invention may include a pharmaceutically acceptable salt of the compound of Formula 1 or Formula 2.

The pharmaceutically acceptable salt of the compound of Formula 1 or Formula 2 may be an acid addition salt formed using an organic acid or an inorganic acid. The organic acid may be, for example, formic acid, acetic acid, propionic acid, lactic acid, butyric acid, But are not limited to, hydrochloric acid, hydrobromic acid, hydrofluoric acid, hydrofluoric acid, malic acid, maleic acid, malonic acid, fumaric acid, succinic acid monoamide, glutamic acid, tartaric acid, oxalic acid, citric acid, glycolic acid, glucuronic acid, ascorbic acid, benzoic acid, phthalic acid, Benzoic acid, benzenesulfonic acid, p-toluenesulfonic acid and methanesulfonic acid-based salts, and the inorganic acid includes, for example, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid and boric acid-based salts. Preferably in the hydrochloride or acetate form, more preferably in the hydrochloride form.

The above-mentioned acid addition salts are prepared by a) directly mixing the compound of formula (1) or (2) and an acid, or b) dissolving and mixing one of them in a solvent or a water solvent, or c) Are placed in an acid in a solvent or an undercooling solvent, and they are mixed.

In addition to the above, additionally saltable forms include, but are not limited to, the salts of gabapentin, pregabalin, nicotinate, adipate, hemimarate, cysteine, acetylcysteine, methionine, arginine, Aspartate and the like.

When the compound of formula (1) or (2) of the present invention is used as a medicine, it may further contain one or more active ingredients showing the same or similar functions. For example, it may contain known skin wrinkle improvement, elasticity enhancement, skin whitening and antioxidant components. The skin wrinkle improvement, the elasticity enhancement, the skin whitening and the antioxidant effect of the composition of the present invention may be further enhanced if the skin wrinkle improvement, the skin elasticity enhancement, the skin whitening, and the antioxidant component are included. When the above ingredients are added, skin safety, easiness of formulation, and stability of effective ingredients can be considered according to the combined use. In one embodiment of the invention, the composition comprises an antioxidant component known in the art, such as tocopherol, selenium, vitamin C and a phenolic compound, kojic acid A substance inhibiting tyrosinase enzyme activity such as arbutin, hydroquinone, L-ascorbic acid and derivatives thereof, and various plant extracts. May further comprise more than one species of component. The additional ingredients may be included in an amount of 0.0001% to 10% by weight based on the total weight of the composition, and the content ranges may be adjusted according to requirements such as skin safety, ease of formulation of the compound of Formula 1 or Formula 2 It will be possible.

In addition, the composition for improving skin wrinkles, improving elasticity, skin whitening, and antioxidant pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier.

Pharmaceutically acceptable carriers may contain a variety of ingredients such as buffer, injectable sterile water, normal saline or phosphate buffered saline, sucrose, histidine, salts and polysorbates.

The pharmaceutical composition of the present invention can be administered orally or parenterally, and can be administered in the form of a general pharmaceutical preparation, for example, various forms of oral and parenteral administration at the time of clinical administration. In the case of formulation, , An extender, a binder, a wetting agent, a disintegrant, a surfactant, and the like.

Solid form preparations for oral administration include tablets, pills, powders, granules, capsules and the like, which may be formulated by incorporating into the pharmaceutical composition of the present invention at least one excipient such as starch, calcium carbonate, Sucrose, lactose, gelatin, and the like.

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

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

In the present invention, the term " wrinkle-reducing effect " refers to inhibiting or inhibiting the generation of wrinkles on the skin, or alleviating already-generated wrinkles.

In the present invention, the 'elasticity-enhancing effect' refers to an increase in elasticity to the skin, which suppresses or inhibits the loss of elasticity of the skin, or alleviates the already-reduced elasticity.

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

In the present invention, the term 'antioxidative effect' refers to the action of free radicals or reactive oxygen species (ROS), which are highly reactive according to oxidative stress caused by intracellular metabolism or ultraviolet rays, Refers to inhibition of oxidation, and includes removal of free radical or reactive oxygen species, thereby reducing damage to the cells.

When the pharmaceutical composition of the present invention contains an effective amount of the compound of Chemical Formula 1 or Chemical Formula 2, it can provide desirable skin wrinkles, elasticity enhancement, skin whitening and antioxidative effects. In the present invention, the term "effective amount" means the amount of a compound capable of promoting regeneration of damaged skin, improving wrinkles, improving elasticity, exhibiting antioxidative and whitening effects. The effective amount of the compound of Formula 1 or Formula 2 contained in the composition of the present invention will vary depending on the form in which the composition is commercialized, the manner in which the compound is applied to the skin, and the time on the skin. For example, when the composition is commercialized as a pharmaceutical product, the compound of Formula 1 or Formula 2 may be contained at a higher concentration than the cosmetic product that is routinely applied to the skin. Therefore, the daily dosage is 0.1 to 100 mg / kg, preferably 30 to 80 mg / kg, more preferably 50 to 60 mg / kg, based on the amount of the compound of formula (1) It can be administered 1 to 6 times a day.

The pharmaceutical 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 present invention can also be provided as a composition for improving skin wrinkles, improving elasticity, skin whitening, and antioxidant dermatological external preparations containing the compound of Formula 1 or Formula 2 as an active ingredient.

When the compound of formula (1) or (2) is used as an external preparation for skin, it may further contain at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softeners, antioxidants, suspending agents, stabilizers, foaming agents, , Surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, lipophilic or lipophilic active agents, And any other ingredients conventionally used in external preparations. The components can also be introduced in amounts commonly used in the field of dermatology.

When the compound of Formula 1 or Formula 2 is provided as an external preparation for skin, it may have a formulation such as, but not limited to, an ointment, a patch, a gel, a cream or a spray.

The present invention can also provide a composition for improving skin wrinkles, improving elasticity, skin whitening, and an antioxidant cosmetic composition containing the compound of Formula 1 or Formula 2 as an active ingredient.

When the compound of formula (1) or (2) is used as a cosmetic product, the cosmetic product containing the compound of formula (1) or (2) as an active ingredient may be prepared in the form of a general emulsified formulation and a solubilized formulation. For example, creams, essences, cosmetic creams, sprays, gels, packs, sunscreens, make-up bases, liquids such as lotions such as lotion, facial lotion, body lotion, A powder, a cleansing lotion, a makeup removing agent such as a cleansing oil, a cleansing foam, a soap, a body wash, and the like.

The cosmetic composition may further contain, in addition to the compound of Formula 1 or Formula 2, a lipid, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, Preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics, in addition to the active agents, water, ionic or nonionic emulsifiers, fillers, sequestering agents and chelating agents And any other ingredients used in the cosmetics field.

In the case of a wash-off type cosmetic such as a make-up remover or a detergent in which the active ingredient remains on the skin in a short period of time when the cosmetic product is made into a product of the formula 1 or the formula 2, Or a compound of formula (2). On the other hand, in the case of leave-on type cosmetics, such as lotion, cream, essence, etc., in which the active ingredient remains on the skin for a long period of time, May also be included. In one embodiment of the present invention, the composition may contain 0.0001 wt% to 10 wt% (preferably 0.0001 wt% to 1 wt%) of the compound of Formula 1 or Formula 2 based on the total weight of the composition. As shown in FIG. When the composition of the present invention contains less than 0.0001% by weight of the compound of formula (1) or (2), sufficient skin wrinkle improvement, elasticity enhancement, whitening and antioxidative effect can not be expected. It is to prevent the possibility of unwanted reactions such as allergies or skin safety problems.

The present invention also relates to a skin wrinkle improving, elasticity enhancing, skin whitening and antioxidant health food containing the compound of the above formula (1) or (2).

The term "health food" as used herein refers to a food prepared by adding the compound of Formula 1 or Formula 2 to a food material such as beverage, tea, spice, gum, confectionery, or the like, or by encapsulation, powdering, However, unlike general medicines, it has the advantage that there are no side effects that can occur when a drug is taken for a long time by using the food as a raw material.

Since the health food of the present invention thus obtained can be taken on a daily basis, it can be expected to have a high skin wrinkle, a skin elasticity, a skin whitening effect, and an antioxidant effect, which is very useful.

When the compound of formula (1) or (2) is used as a food additive, the compound of formula (1) or (2) may be added as it is or may be used together with other food or food ingredients. The amount of the active ingredient to be mixed can be suitably determined according to its intended use (prevention, health or therapeutic treatment). Generally, the composition of the present invention is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight, based on the raw material, when the food or beverage is produced. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, 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 exceeding the above range .

There is no particular limitation on the kind of the food. Examples of the food to which the above substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The above-mentioned natural carbohydrates are sugar saccharides such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio 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, the health food of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, , A carbonating agent used in carbonated drinks, and the like. In addition, the health food of the present invention may contain flesh for the production of natural fruit juice, fruit juice drink and vegetable drink. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

The present invention also provides a composition for promoting stem cell activity comprising the compound of Formula 1 or Formula 2 as an active ingredient.

The term " stem cell " in the present invention is a cell capable of cell division by itself and capable of differentiating into a very specific type of specific cell type. The type of such stem cells is not particularly limited, and in one embodiment, the stem cells may be dermal stem cells. The term " dermal stem cell " means a stem cell capable of differentiating into cells constituting the skin (epidermis, dermis and subcutaneous fat layer). The cells that make up the skin include keratinocytes, melanocytes, and fibroblasts (mainly responsible for biosynthesis of collagen and elastin) present in the epidermis.

The kind of the skin stem cell is not particularly limited. The dermal stem cells used in the present invention can be used irrespective of where they originate from. For example, dermal stem cells may be obtained from a known source of dermal stem cells, e. G., From the basal layer of hair follicles 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, a mouse, a rat, a guinea pig, a rabbit, a monkey, a pig, a horse, a cattle, a sheep, Preferably the mammal can be human. Such methods of obtaining dermal stem cells from dermal stem cell sources are well known in the art.

In the present invention, the term " stem cell activation promoting effect " refers to a stem cell growth promoting effect.

Dermal stem cells play an important role in maintaining the health and physiological and biochemical homeostasis of the skin. Stem cells present in the skin are also abnormally functioned due to the effect of aging, and thus various problems arise as the homeostasis of the skin is broken. Through the activity of these stem cells, various phenomena of skin aging can be improved [Non-Patent Document 5]. In the present invention, skin regeneration, wrinkle improvement, elasticity enhancement, whitening and antioxidative effect were confirmed.

Accordingly, the present invention can provide a pharmaceutical composition, an external preparation for skin, a health food and / or a cosmetic composition containing the stem cell activity promoting composition. Such pharmaceutical composition, external preparation for skin, health food and cosmetic preparation are the same as those described in the description of pharmaceutical composition, external preparation for skin, health food and cosmetic formulation containing the compound of formula (1) or (2).

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

[ Example ]

Reference Example  1: Material information

[Chemical Formula 1]

Material name: 5α-cholestan-3β, 6α, 7α, 8,15α, 16β, 26-heptol

CAS No. : 100101-16-4

 (2)

Material name: 5α-cholestan-3β, 4β, 6α, 7α, 8,15β, 16β, 26-

CAS No. : 128298-63-5

Reference Example  2: star starfish extract and Fraction  Produce

(1) Starfish extraction and preparation of its solvent fraction

Starfish ( Asterina About 4 kg of pectinifera was added and 15 L of methanol was added. The mixture was extracted twice at room temperature. The filtered material was dried under reduced pressure to obtain a methanol extract. The methanol extract was suspended in 1 L of distilled water and sequentially fractionated with dichloromethane (51.8 g), ethyl acetate (0.19 g), and n- butanol (10.1 g) ≪ / RTI >

(2) Isolation of starch extract from active ingredients

The dichloromethane fraction of the compound (1) was subjected to a silica gel column and eluted by gradually changing the composition of dichloromethane / methanol = 10/1 to 0/100 (v / v) to obtain 10 small fractions.

Compounds 1 and 2 were separated by fractionating 3 (1.7 g) with ODS column using 70% methanol as elution solvent.

The separated compounds 1 and 2 were identified by NMR and MS spectroscopic methods, and their structure was determined by comparing with the previously reported articles.

5α-cholestan-3β, 6α, 7α, 8,15α, 16β, 26-

White amorphous powder; ^{1 H-NMR (600 MHz,} methanol- d 4) δ: 0.93 (3H, d, J = 6.6 Hz, H-21), 0.95 (3H, d, J = 6.6 Hz, H-27), 1.02 (3H , 3.71 (1H, d, J = 10.5, 6 Hz, H-26), 3.51 (1H, m, H-3), 3.78 2H, overlapped, H-6,7), 4.01 (1H, dd, J = 7.2, 1.8 Hz, H-16), 4.15 (1H, dd, J = 11.1, 1.8 Hz, H-15); ^{13 C-NMR (150 MHz,} methanol- d 4) δ: 14.0 (C-19), 17.0 (C-18), 17.5 (C-27), 18.4 (C-21), 19.4 (C-11), 25.0 (C-23), 30.7 (C-20), 31.6 (C-2), 32.4 (C-10), 39.5 (C-1), 43.2 (C-12), 44.6 (C-5), 45.6 (C-13), 51.3 C-17), 68.6 (C-26), 69.0 (C-6), 72.4 (C-3), 76.5 -16); EIMS m / z (rel. Int.,%): 484 [M] ⁺ (1.1), 466 [MH ₂ O] ⁺ (7.6).

Tetrahedron, 38, 3615-3622 (1982)

5α-cholestan-3β, 4β, 6α, 7α, 8,15β, 16β, 26-

White amorphous powder; ^{1 H-NMR (600 MHz,} methanol- d 4) δ: 0.93 (3H, d, J = 6.6 Hz, H-27), 0.97 (3H, d, J = 6.6 Hz, H-21), 1.18 (3H H-19), 1.26 (3H, s, H-18), 3.45 (1H, dd, J = 10.2, 5.4 Hz, H-26), 3.47 1H, d, J = 3 Hz , H-7), 4.21 (1H, m, H-4), 4.24 (1H, t, J = 7.2 Hz, H-16), 4.30 (1H, dd, J = 11.7 , 3 Hz, H-6), 4.50 (1H, t-like, J = 5.4 Hz, H-15); ^{13 C-NMR (150 MHz,} methanol- d 4) δ: 17.0 (C-19), 17.5 (C-27), 17.9 (C-18), 18.6 (C-21), 18.8 (C-11), 25.0 (C-23), 26.4 (C-2), 31.1 (C-20), 35.1 (C-24), 37.2 (C-12), 44.5 (C-13), 47.9 (C-5), 51.4 (C-9), 55.4 C-26), 69.7 (C-4), 71.2 (C-15), 72.9 (C-16), 73.9 (C-3), 77.2 (C-7), 79.2 (C-8); EIMS m / z (rel. Int.,%): 500 [M] ⁺ (0.8), 482 [M - H ₂ O] ⁺ (6.9).

Liebigs Ann. Chem., 12 , 1185-1189 (1988)

Nat. Prod. Res., 27 , 1816-1822 (2013)

Reference Example  3: Culture of dermal stem cells

Human epidermal stem cells purchased from Cellntec were added to 48-well plates (6 × 10 ³ cells / well), to which BPE (bovine pituitary extract) similar to fetal bovine serum was added And cultured for 24 hours at 5% CO ₂ and 37 ° C using CNT-57 medium (Cellntec). Thereafter, the culture solution was removed through a suction tube, and then treated with 2 ppm of each of Chemical Formulas 1 and 2 in the CNT-57 medium from which BPE had been removed, followed by culturing at 5% CO ₂ and 37 ° C for 72 hours.

Experimental Example  One: CCK Promoting the proliferation of skin stem cells through the -8 evaluation method

The skin stem cells cultured in Reference Example 3 were evaluated for CCK-8 (cell counting kit-8). The CCK-8 evaluation is an indirect method of measuring the density of living cells by measuring the absorbance of formazan produced by decomposition of tetrazolium salt in the dehydrogenase in the intracellular electron transport system.

Cells were spectrophotometrically measured by treating CCK-8 solution (treated with 1/10 of the medium volume) at 37 ° C for 2 hours and then measuring the absorbance at 450 nm. The proliferation rate (%) of the compounds of the formulas (1) and (2) was determined from the measured absorbance value by the following equation (1) with respect to the control group (CNT-57 medium supplemented with BPE) 1.

[Equation 1]

Growth rate (%) = (absorbance of sample-treated group / absorbance of control group) x 100

Promoting the proliferation of dermal stem cells sample density Cell proliferation (%) versus control The compound of formula (1) 2 ppm 131% The compound of formula 2 2 ppm 124% Control group - 100%

As can be seen from the results of Table 1, the dermal stem cells in the medium treated with the compound of formula (I) or (II) of the present invention exhibited an excellent proliferation promoting effect.

Experimental Example  2: Elastase  Active inhibitory effect

The activity inhibition effect of elastase, an enzyme that degrades elastin, was confirmed as follows.

Elastase was derived from human leukocyte cells and MeOSuc-Ala-Ala-Pro-Val-pNA, a synthetic substrate for elastase, was used. The buffer solution used was 100 mM Tris (pH 7.5) solution. Elastase was finally used at 0.2 mU using a buffer solution. The synthetic substrate of elastase was diluted with buffer to make a final concentration of 0.5 mM after making 100 mM solution using DMSO. At this time, the positive control group was set as quercetin, which is known as an inhibitor of elastase, at a concentration of 10 ppm. Elastase inhibition candidates were added to a final concentration of 10 ppm. The reaction was carried out in a 96-well plate and allowed to react at room temperature for 20 minutes. Absorbance was measured at 405 nm using a spectrophotometer at intervals of 1 minute, and the slope of the absorbance versus time was determined to determine the activity of the enzyme. The inhibition rate of elastase was calculated by the following formula (2).

&Quot; (2) "

Elastase inhibition rate (%) = ((slope of control - slope of sample) / slope of control) x100

Elastase inhibitory effect (number of repeats = 3) sample Enzyme activity Inhibition rate (%) The compound of formula (1) (10 ppm) 4.3 63.87 The compound of formula (2) (10 ppm) 5.2 56.30 The positive control (quercetin, 10 ppm) 3.5 70.59 The control (DMSO, 10 ppm) 11.9 -

As can be seen from the results in Table 2, when the compound of Chemical Formula (1) or (2) was treated, the effect was less than that of the positive control, but the effect of inhibiting the activity of elastase was exhibited.

Experimental Example  3: Promoting collagen synthesis

Compounds of formulas (1) and (2) were added to the culture medium of human-derived fibroblasts to examine the promoting effect of collagen synthesis at the cellular level.

The increased amount of collagen was quantitated using a PICP EIA kit (procollagen type I C-peptide enzyme immunoassay KIT). The method of evaluating cytotoxicity by varying the concentration (ppm) of the compound of Chemical Formula (1) or Chemical Formula (2) in the human-derived fibroblasts before the experiment (MTT test by culturing fibroblasts [Reference: Mossman T. (1983). To evaluate the extent of increase in the total amount of collagen, a concentration without cytotoxicity was selected (less than 10 ppm) .

Samples were added to the culture medium of human fibroblasts and cultured for 1 day. Then, the culture solution was taken and the degree of increase in total amount of collagen at each concentration was measured at 450 nm using a PICP EIA kit using a spectrophotometer. For the comparison of the effects, the increase of the total amount of collagen was measured by the same method for the culture medium of fibroblasts to which nothing was added (control group) and the sample added with vitamin C to a final concentration of 52.8 ppm. The total amount of collagen was measured as UV absorbance. The increase rate of the total amount of collagen was calculated by the ratio of the total amount of collagen relative to the control, and the effect of increasing the total amount of collagen (number of repeats = 3) is summarized in Table 3 below.

Increase effect of total collagen (number of repeats = 3) sample Absorbance average Collagen growth rate (%) The compound of formula (1) (10 ppm) 554 121 The compound of formula (2) (10 ppm) 583 128 The positive control (vitamin C, 52.8 ppm) 571 125 Control group (no addition) 456 -

As can be seen from the results of Table 3, the compound of Chemical Formula (1) or (2) showed an effect of promoting collagen synthesis and increasing the total amount of collagen.

Experimental Example 4: Whitening effect - Confirmation of melanin formation inhibitory effect

Each of the compounds of formulas (1) and (2) was added to the culture medium of rat B-16 mouse melanoma cells to determine the whitening effect by measuring the total amount of melanin at the cellular level (Lotan R., Lotan D. Cancer Res. 40: 3345-3350, 1980).

To evaluate the toxicity of melanoma cells in rats before the experiment, whitening evaluation was performed by selecting a concentration that is not toxic.

Each of the compounds of Formulas (1) and (2) was added to the culture medium to a final concentration of 10 ppm, and the control group, arbutin, was added to the medium so as to have a concentration of 100 ppm, which was then treated with B-16 melanoma cells and cultured for 3 days.

Cells were then trypsinized, detached from the culture, centrifuged, and extracted with melanin. The removed cells were incubated with 1 ml of sodium hydroxide solution (1N), boiled for 10 minutes to dissolve melanin, and the absorbance was measured at 400 nm using a spectrophotometer to determine the amount of melanin produced.

The amount of melanin was measured by an absorbance of 1 × 10 ⁶ cells per unit cell, and the total amount of melanin production relative to the control group was calculated as the inhibition rate (%). The results are summarized in Table 4 below.

Inhibitory effect of melanin formation at the cellular level (number of repeats = 3) sample Melanin production (abs) Inhibition rate (%) The compound of formula (1) (10 ppm) 0.218 33.93 The compound of formula (2) (10 ppm) 0.247 25.15 Positive control (arbutin, 100 ppm) 0.228 30.91 The control (DMSO, 10 ppm) 0.33 -

As can be seen from the results of Table 4, the compound of Chemical Formula (1) or (2) showed excellent whitening effect even at a lower concentration than arbutin, which is a positive control.

Experimental Example  5: Antioxidant effect - Free radical  Erasure rate

The free radical scavenging activity was measured to confirm the antioxidative activity of each of the compounds of the formulas (1) and (2). Free radical scavenging activity was measured using DPPH. DPPH is a relatively stable free radical, which shows maximum absorption at 517 ㎚ in the presence of radicals and loses its absorbance when radicals are eliminated. DPPH was purchased from Sigma Co., Ltd. (USA) and dissolved in methanol at a concentration of 0.15 mM.

First, 100 μl of the compound of Formula 1 or 2 and ascorbic acid were added to each well of a 96-well plate. 100 μl of DPPH solution was added thereto, and the mixture was allowed to stand at room temperature for 30 minutes. Then, the absorbance at 517 nm was measured using a Microplate reader (BioTek EL-340). At this time, methanol was added instead of the sample to give a control group.

Free radical scavenging activity and the results are obtained for the IC ₅₀ half maximal inhibitory concentration (median inhibition) shown in Table 5. IC ₅₀ is a common method of expressing the free radical scavenging ability as a concentration of ascorbic acid and the compound of formula (I) or (II) required to remove 50% of the free radicals of the no-added control group.

Free radical scavenging ability sample IC ₅₀ (%) The compound of formula (1) 0.0091 The compound of formula 2 0.0075 Ascorbic acid 0.0007

As shown in Table 5, the compound of Formula 1 or Formula 2 of the present invention has lower free radical scavenging ability than ascorbic acid.

Formulation example  1: Preparation of pharmaceutical preparations

1. Preparation of tablets

0.2 mg of the compound of formula (1) or (2)

100 mg of corn starch

100 mg of milk

2 mg of magnesium stearate

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

Formulation example  2: Manufacture of cosmetics

1. Manufacture of nutritional cream

A nutritional cream containing the compound of formula (1) or (2) as an active ingredient was prepared according to a conventional method, as shown below.

0.2% by weight of the compound of formula (1) or (2)

Beta-1,3-glucan 5.0 wt%

Wax 10.0 wt%

Polysorbate 60 1.5 wt%

≪ tb > < tb > < tb >

0.5% by weight of sorbitan sesquioleate

Liquid paraffin 10.0 wt%

Squalane 5.0 wt%

Caprylic / capric triglyceride 5.0 wt%

Glycerin 5.0 wt%

3.0% by weight of butylene glycol

3.0% by weight of propylene glycol

0.2% by weight triethanolamine

Preservative 0.05 wt%

0.05% by weight of pigment

0.05% by weight fragrance

Purified water to 100%

Formulation example  3: Topical  Produce

1. Manufacture of ointment

An ointment containing the compound of formula (1) or (2) as an active ingredient was prepared according to a conventional method, as follows.

0.5% by weight of the compound of formula (1) or (2)

Beta-1,3-glucan 10.0 wt%

Wax 10.0 wt%

Polysorbate 60 5.0 wt%

≪ tb > < tb > < tb >

0.5% by weight of sorbitan sesquioleate

Vaseline 5.0 wt%

Liquid paraffin 10.0 wt%

Squalane 5.0 wt%

SHARE BUTTER 3.0 wt%

Caprylic / capric triglyceride 5.0 wt%

Glycerin 10.0 wt%

Propylene glycol 10.2 wt%

0.2% by weight triethanolamine

Preservative 0.05 wt%

0.05% by weight of pigment

0.05% by weight fragrance

Purified water to 100%

Claims (7)

delete A cosmetic composition for enhancing skin elasticity or improving wrinkles comprising a compound represented by the following formula (1) as an active ingredient.
[Chemical Formula 1]

A cosmetic composition for skin whitening comprising a compound represented by the following formula (1) as an active ingredient.
[Chemical Formula 1]

1. An antioxidant cosmetic composition comprising a compound represented by the following formula (1) as an active ingredient.
[Chemical Formula 1]

5. The method according to any one of claims 2 to 4,
Wherein the cosmetic composition is promoted stem cell activity.
5. The method according to any one of claims 2 to 4,
Wherein the compound represented by Formula 1 is extracted from starfish.
5. The method according to any one of claims 2 to 4,
Wherein the active ingredient is contained in an amount of 0.00001 to 10% by weight based on the total weight of the composition.