KR20170025371A - Composition for improving skin - Google Patents

Abstract

The present invention relates to a composition for improving skin. The balloon orchid extract according to the present invention is effective for promoting proliferation of skin stem cells, promoting collagen synthesis of skin fibroblasts, promoting melanin synthesis, free radical scavenging effect, inhibiting NO production, excellent effect of PPAR alpha, Cosmetics or food for improving skin regeneration, wrinkle improvement, elasticity enhancement, antioxidant, anti-inflammation, atopic improvement, skin moisturization and skin texture improvement through the effect of improving skin texture and improving skin texture. In addition, it promotes the proliferation of dermal stem cells and exhibits a stem cell-retaining effect, and thus can be used for the production of a cosmetic for promoting stem cell activity.

Description

[Composition for improving skin]

The present invention relates to a skin improving composition exhibiting skin regeneration, wrinkle improvement, elasticity enhancement, antioxidation, anti-inflammation, atopic improvement, skin moisturizing, skin texture improvement and stem cell activity promoting effect.

Collagen is a major substrate protein produced in fibroblasts of the skin and exists in extracellular epilepsy. Its important functions are mechanical rigidity of skin, resistance of connective tissues and binding force of tissues, support of cell adhesion, division of cells and differentiation Growth or wound healing) are known. Such collagen is reduced by aging and photo aging caused by ultraviolet irradiation, which is known to be closely related to the wrinkling of the skin. Also, in recent years, extensive research on skin aging has developed, and important functions of collagen in skin have been revealed.

Effective ingredients promoting collagen synthesis and exhibiting wrinkle-reducing effects are known. For example, retinoic acid, transforming growth factor (TGF) [non-patent document 1], animal placenta-derived protein [Patent document 1], betulinic acid [Patent document 2], chlorella extract [ Patent Documents 3 and 4] are known as collagen synthesis promoting substances. However, the above-mentioned effective ingredients are limited in the use amount due to safety problems such as irritation and redness when applied to the skin, or have insufficient effect, so that the effect of improving the skin function by promoting the collagen synthesis of the skin can not be expected.

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. Antioxidants are widely distributed in copper and plants. Many phenolic compounds, flavonoids, tocopherols, vitamin C and selenium are known in fruits and vegetables. However, the antioxidant substances present in nature can not be expected to have practically sufficient effects in skin application. Therefore, although synthetic antioxidants having excellent antioxidant ability and low cost are widely used, their use is restricted due to safety concerns such as human side effects.

In addition, inflammation is an immune response of a human body in response to a wound or disease, and oxidative stress such as ultraviolet rays, active oxygen, free radicals, etc. activate inflammatory factors and cause aging of various diseases and skin. The vasoactive polypeptide, kinin, plasmin and complement, have vasodilatory and contraction and chemotaxis effects, as well as lymphokines such as interleukin-6 (IL-6) Phosphorus and arachidonic acid are responsible for inflammation. Arachidonic acid is metabolized through inflammatory mediators such as prostaglandin and lukotrienes via two pathways: cyclooxygenase or lipooxygenase, mediating a variety of inflammatory responses.

On the other hand, in order to eliminate inflammation, elimination of inflammation source, reduction of vital reaction and symptoms is called anti-inflammatory. To date, substances used for antiinflammatory purposes include flutenamic acid, ibuprofen, benzydamine, indomethacin, and steroids, prednisolone, , Dexamethasone, and the like. It is also known that allantoin, azene, hydrocortisone and the like are effective for antiinflammation. However, these materials are limited in their use due to safety of skin, .

In addition, the epidermis located at the outermost part of the skin protects against various external physical, chemical and mechanical stimuli and protects against excessive divergence of body water through the skin. This protective function is possible by normally forming and maintaining the stratum corneum composed of keratinocytes. The keratinocyte is a cell formed by a stepwise change in morphology and function while a basal cell that continuously proliferates in the stratum basale moves to the stratum corneum, Forming cells are removed from the skin and the new keratinocytes from the epidermis's bottom layer repeat the process of epidermis differentiation or keratinization replacing its function. In this keratinization process, keratinocytes produce intercellular lipids such as natural moisturizing factors (NMF) and ceramides, cholesterol, and fatty acids, which act as barrier layers to the outside, As shown in Fig.

In addition, skin dryness, which is considered to be one of the major diseases of modern society, is one of the symptoms caused by skin barrier function abnormality. Recently, environmental pollution, increase in dry environment such as apartments and high rise building, increase in social stress, Of excessive bathing, skin aging, and the like, and the cases of severe symptoms and the need for treatment are also increasing steadily.

Atopic dermatitis, which is present in 10% of children in recent years, is also known to be a cause of dry skin syndrome, and more fundamentally, abnormal skin barrier function. In order to treat such atopic dermatitis, studies have been carried out in order to supply water from the outside or minimize water loss from the body by focusing on proper moisture retention in the skin in the past. Actually, ceramide or derivatives thereof have been developed and are widely used in the pharmaceutical or cosmetic field.

However, the use of such a moisturizing agent is not only a fundamental treatment but a temporary symptom relief, and thus has not shown sufficient effect for treatment of dry skin and skin barrier function including atopic dermatitis. Therefore, it is urgent to develop a substance that can restore the damaged skin barrier fundamentally.

Recently, it has been known that PPAR alpha (peroxisome proliferator activated receptor-alpha) present in keratinocyte cells is activated by binding to its ligand, thereby promoting the differentiation of keratinocytes and reconstructing damaged skin barrier. In the case of applying Clofibrate (non-patent document 2) or WY 14643 (non-patent document 3), which is an agonist of known PPAR alpha, to the skin of which skin barrier is damaged, differentiation of keratinocytes is promoted And recovery of the skin barrier is promoted.

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 according to age and site. The skin texture is divided into primary (about 20-100 μm), secondary (about 5-40 μm), and tertiary (about 0.5 μm) lines depending on the depth of the line. And has a distinctive polygonal shape and star formation due to the contact of the lines [Non-Patent Document 4].

However, as the age increases or the skin becomes damaged, the network structure of the dense fine lines collapses and fine lines (secondary, tertiary, etc.) disappear and the primary lines become deeper and wrinkles are known to be produced 5,6]. In other words, wrinkles, a typical sign of skin aging, are accumulations of minute changes that have already occurred over a long period of time.

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. According to the developmental stage, the embryonic stem cells are repeatedly differentiated and become cells with the characteristics of each tissue function. After the embryo, a certain number of stem cells remain in the tissue. . 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 layer of the epidermis. 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 relatively 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 repair of the epithelium after injury [Non-Patent Document 7]. The epithelial stem cells of the skin 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, have. 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.

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. Therefore, it is possible to improve various phenomena of skin aging through the activation of stem cells [Non-Patent Document 8].

In addition, it is possible to provide a skin regenerating method which is more safe than a living body, has a stable active ingredient, and is more effective than a substance having a skin regeneration, wrinkle improvement, elasticity enhancement, antioxidant, antiinflammation, atopic improvement, skin moisturizing, It is urgently required to develop a composition having wrinkle improving, elasticity improving, antioxidant, anti-inflammatory, atopic improvement, skin moisturizing and skin tone improving activity.

On the other hand, balloon orchid is a perennial plant of orchid orchidaceae, and its scientific name is <Calypso bulbosa Reichb.fil.> And it is also called bongboran or ovenulran. Because petals look like balloons, they are called 'balloon orchids'. It is distributed in Korea (Mt. Paektu) and the sub - region including North of Japan. Mainly used as corn plants.

It grows up to 30㎝ in height from the coniferous forest. The rootstock is fleshy and ellipsoidal, with one leaf and stem at the end. Leaves are petiole, egg-shaped or oval-shaped. The tip of the leaf is pointed, the bottom is round, the wrinkles are vertical, and the back side is purplish.

Flowers bloom in May-June, one at the end of peduncle and light pink. The peduncle is 6-15 cm high and has two leaf-like leaves on its lower part. The bract is one and has a wide line. Three calyxes and two petals are similar in shape, narrow and narrow, and 3cm long and light reddish purple.

There are no studies on the skin wrinkles and stem cells of balloon orchids.

1. Japanese Patent Publication No. 8-231370 2. Japanese Patent Publication No. 8-208424 3. Japan Pyeonghwa 9-40523 4. Japan Patent No. 10-36283

1. Cardinale G. et al, Adv. Enzymol., 41, p. 425, 1974 2. Feingold et al., J. Invest. Dermatol., 110, pp 368-375, 1998. 3. Feingold et al., J. Invest. Dermatol., 115, pp 353-360, 2000. 4. Journal of the European Academy of Dermatology and Venereology. 12: 103-114, 1999 5. The British journal of dermatology. 110: 129-138, 1984 6. Skin research and technology.5: 189-194, 1999 7. Epidermal Stem Cells of the Skin, Cedric Blanpain, Elaine Fuchs, Annual Review of Cell and Developmental Biology, November 2006, Vol. 22, Pages 339-373 8. Human skin stem cells and the aging process, Catherine Niemann, Stem Cell Aging and Regenerative Medicine, November 2008, Pages 986-997

The present inventors have found that the balloon orchid extract promotes the proliferation of skin stem cells, promotes skin regeneration, promotes collagen synthesis of fibroblasts of the skin, thereby improving wrinkles, promoting elasticity, eliminating free radicals, exhibiting antioxidative effects, Inhibition of NO production, anti-inflammatory effect, excellent activity of PPAR alpha, improvement of atopy, improvement of skin moisturization due to increased expression of filar green, improvement of skin texture due to improvement of skin brightness, Promoting proliferation of cells, exhibiting a stem cell-maintaining effect, and confirming the effect of promoting stem cell activity, thereby completing the present invention.

Accordingly, an object of the present invention is to provide a composition for improving skin regeneration, wrinkle improvement, elasticity enhancement, antioxidation, antiinflammation, atopic improvement, skin moisturizing and skin texture improvement comprising a balloon orchid extract as an active ingredient.

Another object of the present invention is to provide a composition for promoting stem cell activity comprising the following inflatable orchid extract as an active ingredient.

As a means for solving the above-mentioned problems, the present invention provides a method for improving the skin regeneration, wrinkle improvement, elasticity enhancement, antioxidant, anti-inflammatory, atopic improvement, skin moisturizing, A composition for improving skin texture is provided.

As another means for solving the above problems, the present invention provides a composition for stimulating stem cell activity comprising, as an active ingredient, a balloon orchid extract for the production of cosmetic formulations.

The balloon orchid extract according to the present invention promotes skin regeneration by promoting proliferation of skin stem cells, promotes collagen synthesis of fibroblasts of skin to improve wrinkles, promotes elasticity, and eliminates free radicals to exhibit antioxidative effects , NO production is inhibited and anti-inflammatory effect is exerted. PPAR alpha activity is excellent, and atopy is improved. In addition, skin moisturization is improved due to an increase in fila green expression and skin texture improvement effect is improved by skin brightness improvement, It can be used for medicine, cosmetics or foods.

In addition, the balloon orchid extract promotes the proliferation of dermal stem cells and exhibits a stem cell-maintaining effect, and thus can be used for the production of cosmetic formulations for promoting stem cell activity.

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

In order to exert an excellent effect when applied to the actual skin, it is necessary to improve the skin regeneration, wrinkle improvement, elasticity enhancement, anti-inflammation, anti-oxidant, anti-inflammation, It has excellent antioxidant, anti-inflammatory, anti-atopic effect, skin moisturizing and skin tone improving activity and is excellent in the ability to permeate through the skin and be absorbed and is excellent in skin regeneration, wrinkle improvement, elasticity enhancement, antioxidant, anti- It is preferable that the composition is low in volatility so as to be able to stay for a sufficient time to exhibit the improvement effect of the composition and that the active ingredient remains stable on the composition or on the skin and is easily prepared into medicines, cosmetics or foods, and is safe for the skin. However, the components satisfying all of the above-mentioned characteristics among the known components are not common. For example, some skin regeneration, wrinkle improvement, elasticity enhancement, antioxidant, antiinflammation, atopic improvement, skin moisturizing and skin tone improving ingredients can be used for skin regeneration, wrinkle improvement, elasticity enhancement, antioxidant, Atopy improvement, skin moisturizing and skin texture improving activity are excellent, but it is difficult to apply to actual skin because of its ability to be absorbed through the skin. Other active ingredients have low hydrophilicity, making them difficult to formulate into medicines, cosmetics, and foods. In addition, some of the skin regeneration, wrinkle improvement, elasticity enhancement, antioxidant, anti-inflammation, atopic improvement, skin moisturizing and skin tone improving ingredients are decomposed or transformed into other compounds when exposed to heat, light, In some cases, the effect disappears before it is applied.

As can be seen from the following examples, the infant orchid extract of the present invention is superior in low density to promoting proliferation of skin stem cells, promoting collagen synthesis, antioxidative effect, antiinflammatory effect, PPAR alpha activity effect, It can be used as an active ingredient of medicines, cosmetics or food compositions for skin regeneration, wrinkle improvement, elasticity enhancement, antioxidation, anti-inflammation, atopic improvement, skin moisturization and skin texture improvement.

Accordingly, the present invention provides a composition for improving skin regeneration, wrinkle improvement, elasticity enhancement, antioxidation, antiinflammation, atopic improvement, skin moisturization and skin texture improvement comprising a balloon orchid extract as an active ingredient.

In the present invention, on the other hand, a balloon orchid is a perennial plant of orchid orchid and its scientific name is Calypso bulbosa Reichb.fil.

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

The balloon orchid extract may be an extract of flowers, leaves, branches, fruits, roots, stems, seeds or a mixture thereof of a balloon orchid.

Preferably, the above-mentioned balloon orchid extract may be an extract obtained by extracting a balloon orchid with water and / or an organic solvent. 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 nonpolar organic solvent may be ether, chloroform, benzene, hexane or dichloromethane. For example, the lower alcohol having 1 to 5 carbon atoms may be methanol, ethanol, propanol, butanol or isopropanol.

In one embodiment, the balloon orchid extract may include a fraction obtained by fractionating the primary extract extracted using the above-described extraction solvent with an extraction solvent having a different polarity. For example, the balloon orchid extract may be a fraction obtained by extracting with an alcohol having 1 to 5 carbon atoms and then fractionating again with a solvent having a different polarity such as ether, benzene, hexane or the like. Two or more kinds of solvents may be used for the fractionation, and the solvent extracts may be sequentially used or mixed according to the polarity of the solvent.

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

The fractions obtained by performing the above-described extract or the fractionation process can be further purified by silica gel column chromatography, thin layer chromatography or high performance liquid chromatography Further purification can be achieved by purification using various chromatographic techniques.

The compositions of the present invention may be used for the manufacture of pharmaceutical formulations.

The pharmaceutical formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules.

In addition, the composition may further contain one or more active ingredients showing the same or similar functions. For example, it may contain known skin regeneration, wrinkle improvement, elasticity enhancement, antioxidant, anti-inflammatory, atopic improvement, skin moisturizing and skin tone improving ingredients. When the composition of the present invention contains the components of skin regeneration, wrinkle improvement, elasticity enhancement, antioxidation, anti-inflammation, atopic improvement, skin moisturizing and skin texture improvement, skin regeneration, wrinkle improvement, elasticity enhancement, antioxidation, anti- , The skin moisturizing effect and the skin texture improving effect can be further enhanced. 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 present invention, the composition is a skin regeneration component known in the art comprising retinoic acid, TGF, protein from animal placenta, betulinic acid and chlorella extract, antioxidant components known in the art, such as tocopherol, selenium , Vitamin C and phenolic compounds, whitening ingredients known in the art, substances inhibiting the activity of tyrosinase enzymes such as kojic acid and arbutin, hydroquinone, vitamin C (L- Ascorbic acid, derivatives thereof, and various plant extracts. The additional ingredient may be contained in an amount of 0.0001 to 10% by weight based on the total weight of the composition, and the content may be adjusted according to requirements such as skin safety, ease of formulation of the infant orchid extract.

In addition, the 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, and the like.

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

Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like. These solid preparations can be prepared by mixing the pharmaceutical composition of the present invention with at least one excipient such as starch, calcium carbonate, Sucrose, lactose, gelatin and the like can be prepared.

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

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

In the present invention, the term &quot; wrinkle-reducing effect &quot; 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 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.

In the present invention, the term 'anti-inflammatory effect' refers to inhibition of inflammation. The inflammation is one of defensive responses of biological tissues to certain stimuli, and involves three types of tissue degeneration, circulatory disorder, exudate, and tissue proliferation Complex lesions. More specifically, inflammation is part of congenital immunity and, like in other animals, human congenital immunity recognizes a pattern of cell surfaces that are specifically present in a pathogen. Phagocytes recognize cells with such surfaces as non-magnetic and attack pathogens. If pathogens break through the physical barriers of the body, an inflammatory reaction occurs. Inflammation is a nonspecific defense that creates hostile environments for microorganisms entering the wound. In the inflammatory response, white blood cells that are responsible for the early stage of immune response, when wounded or infected, enter the body to express cytokines. Therefore, the expression level of intracellular cytokine is an index of inflammatory response activation. Examples of skin diseases associated with inflammation include atopic dermatitis, psoriasis, radiation, chemical substances, inflammatory diseases caused by burns, acid burns, vesicular dermatosis, visceral type diseases, itching due to allergies, seborrheic eczema, rose acne, Inflammatory hair loss such as pemphigus vulgaris, polymorphous exudative erythema, erythema nodosum, erythematosus, pharyngitis, alopecia areata, skin T-cell lymphoma, and the like.

In the present invention, the term 'atopic improvement effect' refers to the effect of preventing and treating skin diseases such as atopy and dry skin, and is to inhibit, inhibit, or alleviate atopic symptoms.

In the present invention, the term 'moisturizing effect' refers to inhibiting or reducing the reduction of moisture in the skin, or increasing the moisture content of the skin to smooth and smooth the surface of the skin.

In the present invention, the term &quot; skin texture improving effect &quot; refers to smoothing the skin surface, imparting gloss, and brightening the skin tone by inhibiting or inhibiting roughness of the skin surface due to aging, stress, .

In the present invention, the term "effective amount" refers to an amount effective for promoting regeneration of damaged skin, improving wrinkles, improving elasticity, exhibiting a whitening effect, inhibiting or alleviating oxidation of cells, Refers to the amount of the extract which can improve the skin dryness such as atopy, or can improve the moisturizing effect and skin texture. When the composition of the present invention contains the effective amount of the above-mentioned infant orchid extract, it is preferable that the skin regeneration effect, wrinkle improvement effect, elasticity enhancement effect, skin whitening effect, antioxidant effect, anti- inflammatory effect, atopic improvement effect, moisturizing effect, Can be provided. The effective amount of the above-mentioned infant orchid extract contained in the composition of the present invention will vary depending on the form in which the composition is commercialized, how the compound is applied to the skin, and the time on the skin. For example, when the composition is commercialized as a pharmaceutical formulation, it may contain the above-mentioned balloon orchid extract at a high concentration as compared with the case where it is commercialized as cosmetics to be applied to skin on a daily basis. Therefore, the daily dosage is 0.1 to 100 mg / kg, preferably 30 to 80 mg / kg, more preferably 50 to 60 mg / kg, and 1 to 6 mg / kg per day based on the amount of the balloon orchid extract. &Lt; / RTI &gt;

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

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

When the balloon orchid extract is used as an external preparation for skin, it may further contain at least one selected from the group consisting of a fatty substance, 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, For example, water, ionic or nonionic emulsifiers, fillers, sequestering agents and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or external preparations for skin And any other ingredients used, such as those commonly used in the field of dermatology. The components can also be introduced in amounts commonly used in the field of dermatology.

When the balloon orchid extract is provided as an external preparation for skin, it may have a formulation such as, but not limited to, ointments, patches, gels, creams or sprays.

In addition, the composition for skin regeneration, wrinkle improvement, elasticity enhancement, antioxidation, anti-inflammation, atopic improvement, skin moisturizing and skin texture improvement of the present invention can be used for the production of cosmetic formulations.

The cosmetic formulation may be in the form of a conventional emulsion 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 balloon orchid extract, a lipid, an organic solvent, a solubilizer, a thickener and a gelling agent, a softener, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, As used herein means any emulsion or emulsion which is commonly used in emulsifying or emulsifying agents, fillers, sequestering and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics Adjuvants commonly used in the cosmetics field, such as other ingredients.

The cosmetic formulation may contain a relatively high concentration of the above-mentioned infant orchid extract in the case of a wash-off type cosmetic such as a make-up remover, a detergent or the like in which an effective ingredient stays on the skin in a short period of time. On the other hand, in the case of leave-on type cosmetics such as lotion, cream, essence and the like in which the active ingredient remains on the skin for a long period of time, the lower concentration of the above-mentioned infant orchid extract is contained It may be possible. In one embodiment of the present invention, but not limited thereto, the composition may contain the balloon orchid extract in an amount of 0.0001% to 10% by weight (preferably 0.0001% to 1% by weight) based on the total composition weight have. When the composition of the present invention contains less than 0.0001% by weight of the above-mentioned balloon orchid extract, sufficient skin regeneration, wrinkle improvement, elasticity enhancement, antioxidation, antiinflammation, atopic improvement, skin moisturizing and skin texture improving effect can not be expected. If it is contained in an amount of more than 10% by weight, it is intended to prevent an undesired reaction such as allergy or a problem of skin safety.

In addition, the composition for skin regeneration, wrinkle improvement, elasticity enhancement, antioxidation, anti-inflammation, atopic improvement, skin moisturizing and skin texture improvement of the present invention can be used for manufacturing food formulations.

The food formulation refers to a food prepared by adding the infant orchid extract to a food material such as beverage, tea, spice, gum, confectionery, or the like, or encapsulating, pulverizing, or suspending.

Since the food formulation can be routinely ingested, it is very useful because it can expect high skin regeneration, wrinkle improvement, elasticity enhancement, antioxidation, anti-inflammation, atopic improvement, skin moisturization and skin texture improvement.

When the balloon orchid extract is used as a food additive, the balloon orchid extract may be directly added or used together with other food or food ingredients, and may be suitably used according to a conventional method. 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 foods to which the above substances can be added include dairy products including meats, sausages, breads, chocolates, candies, snacks, confections, pizza, ramen noodles, gums, ice cream, soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

When the food is a beverage, various flavors or natural carbohydrates may be added as an additional ingredient such as a normal drink. The above-mentioned natural carbohydrates are sugar alcohols 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 food formulations may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, And the like. Other food formulations may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. 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 stimulating stem cell activity comprising the above-mentioned infant orchid extract 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 cells' refers to stem cells that can be differentiated 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 'promoting effect on stem cell activity' refers to an effect of promoting stem cell proliferation and / or an effect of maintaining stem cell characteristics, which is a specific indicator molecule and a self-cleaving property of stem cells.

In addition, the compositions of the present invention can be used for the production of cosmetic formulations. These cosmetic formulations are the same as those described in the description of the cosmetic formulation containing the balloon orchid extract.

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.

Manufacturing example  One: Balloon orchid  Preparation of extract

The petals and stems of the balloon orchids were washed and pulverized, and then cooled in a 10-fold volume of ethanol for 5 days, and the extract solution was filtered and concentrated to obtain an extract. To this was added a mixed solution of purified water and ethanol to dissolve and then filtered to prepare a final extract.

Manufacturing example  2: Serum-free Under medium conditions Of dermal stem cells  culture

Human epidermal stem cells purchased from Cellntec were added to 48-well plates (6 × 10 ³ cells / well), supplemented with BPE (bovine pituitary extract) similar to fetal bovine serum And cultured for 24 hours at 5% CO ₂ and 37 ° C using CNT-57 medium (Cellntec). Then, the removal through the medium-introducing tube, and then, PBS solution used to remove the medium components the (GibcoBRL Co.), processes the balloon orchid extract, 0.0005% by weight of the CNT-57 medium that does not include BPE 5% CO _2, and And cultured at 37 ° C for 72 hours.

Experimental Example  One: CCK -8 Through evaluation method Dermal stem cells  Proliferation promoting effect

The skin stem cells cultured in Preparation Example 2 were evaluated for CCK-8 (Cell counting kit-8). The CCK-8 assay is an indirect method of measuring the density of living cells by measuring the absorbance of Formazan, a dehydrogenase in the intracellular electron transport system, produced by the decomposition of tetrazolium salt.

Cells were treated spectrophotometrically 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 stem cell of the balloon orchid extract was determined from the measured absorbance value in accordance with the following equation (1) with respect to the control group (CNT-57 medium supplemented with BPE), and the value thereof is shown in Table 1 below.

[Equation 1]

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

Promoting the proliferation of dermal stem cells Additive sample density Cell proliferation (%) versus control Balloon orchid extract 0.005% (w / w) 145% Control group - 100%

As shown in Table 1, the skin stem cells in the culture medium treated with the balloon orchid extract showed an excellent proliferation promoting effect.

Experimental Example  2: Of dermal stem cells  Stem cell ( stemness ) Confirm maintenance effect

The expression level of p63 was evaluated in order to confirm the stem cell maintenance effect of the dermal stem cells cultured in Production Example 2 above. p63 is an intracellular transcription factor and is well known as a maintenance marker for dermal stem cells. Cells were isolated and cDNA was synthesized. Then, real-PCR was performed with Taqman staining solution to measure the expression level of p63. The concentration of RNA in this experiment was normalized to S16 ribosomal RNA.

The experimental results are shown in Table 2 below. In Table 2 below, the numerical value of the increase in p63 is a multiple of the control group (CNT-57 medium supplemented with BPE).

Stem cell (stem cell) maintenance effect (number of repeats = 3) Additive sample density p63 Incremental multiple (times) Balloon orchid extract 5 / / mL 1.78 Control group - 1.0

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

Example  1: Promoting collagen synthesis

The effect of promoting collagen synthesis at the cellular level was investigated by adding a balloon orchid extract to the culture medium of human derived fibroblasts. The biocompatible collagen was quantitated using a PICP EIA kit (Procollagen Type I C-Peptide Enzyme Immunoassay Kit).

(7 × 10 ⁴ cells / cm ² ) with vitamin C and a control (no supplement) to a final concentration of 10 μg / mL or 20 μg / mL to obtain a culture medium containing 1 After culturing for one day, the culture broth was taken and the degree of collagen biosynthesis at each concentration was measured at 450 nm using a spectrophotometer with a PICP EIA Kit. The collagen biosynthesis performance was calculated by the relative performance relative to the control group and the results are summarized in Table 3 below.

Promotion of collagen synthesis by concentration (number of repeats = 3) Additive sample Application concentration (/ / ml) Increase in collagen synthesis (%) compared to control Balloon orchid extract 10 [mu] g / ml 28.5% Balloon orchid extract 20 ug / ml 59.7% Vitamin C 50 ug / ml 30.3%

As shown in Table 3, the balloon orchid extract has excellent collagen synthesis ability against human-derived fibroblasts, and in general, a collagen synthesis effect with a lower concentration than that of vitamin C, which is known to have a collagen synthesis ability, is obtained .

Example  2: Antioxidant effect - Free radical scavenging rate

Free radical scavenging activity was measured to confirm the antioxidative activity of the balloon orchid extract. Free radical scavenging activity was measured using DPPH. DPPH was purchased from Sigma Co. (Sigma Co., Ltd, USA) and used. First, a standard DPPH ethanol solution of 1.5 mM (0.06 mg / mL) was prepared. Then, ethanol was added to ascorbic acid, which is an antioxidant, as a reference material, and samples were made at concentrations of 50 ㎍ / mL, 25 ㎍ / mL, 12.5 ㎍ / mL, 6.25 ㎍ / mL and 3.125 ㎍ / mL. Then, the sample and standard DPPH solution were added at the same ratio, stirred well, reacted at 37 ° C for 30 minutes, and absorbance was measured at 520 nm. At this time, ethanol was added instead of the sample to give a control group. IC ₅₀ , which is the half-maximal inhibitory concentration (inhibition value), was determined for the free radical scavenging ability, and the results are shown in Table 4 below. IC ₅₀ is a common method of expressing the free radical scavenging ability as a concentration of ascorbic acid and balloon orchid extract required to remove 50% of the free radicals of the no-added control group.

Free radical scavenging rate (IC ₅₀ ) sample Free radical scavenging rate (ug / ml) Ascorbic acid 5.2 Balloon orchid extract 4.9

As shown in Table 4, the balloon orchid extract has high antioxidative effect compared to ascorbic acid, which is a known antioxidant.

Example  3: Anti-inflammatory effect -NO generation inhibitory effect

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

Specifically, RAW264.7 cells, macrophages of mice, were subcultured several times, placed in 24-well plates at 3 × 10 ⁵ per well, and cultured for 24 hours. Then, the balloon orchid extract was diluted with the concentrations shown in the following Table 6 and replaced with the cell culture medium containing the diluted. At this time, 20 μg / mL of L-NMMA (L-NG-Monomethylarginine), which is an inhibitor of NO production, was treated with a positive control and cultured for 30 minutes. Lipopolysaccharide (LPS) Lt; / RTI &gt; 100 μl of the supernatant was transferred to a 96-well plate, and 100 μl of the GRIESS solution was added thereto at room temperature for 10 minutes. The absorbance at 540 nm was measured to determine the NO inhibitory effect of Compound 1, Table 5 shows the results.

NO generation inhibitory effect sample NO production inhibition rate (%) L-NMMA (positive control) 40.95 Balloon orchid extract (0.1%) 37.24 Balloon orchid extract (1%) 41.74

As shown in Table 5, it can be seen that the balloon orchid extract has the same inhibitory effect on NO production as that of L-NMMA, a known anti-inflammatory substance.

Example  4: PPAR  Verification of Alpha Activation Promotion Effect

In order to confirm the PPAR alpha activation promoting effect of the balloon orchid extract, the following experiment was conducted using a balloon orchid extract. C2C12 cells (ATCC CRL-1772), a mouse myoblast cell line, were subcultured in DMEM (Dulbecco's Modified Eagle's Medium) medium containing 10% fetal bovine serum. As the vector used herein,

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

2) a GAL4 response sequence inserted into a multiple restriction enzyme recognition site of a pGL3 vector (Promega) expressing luciferase and 3) a vector inserted into a transfection internal control with? -Galactosidase β-galactodisase) was used.

The cultured cells were plated on a 24-well plate at a concentration of 4 × 10 ⁴ and cultured for 12 hours. The three types of plasmid genes were transiently transfected using lipopectamine . After 8 hours, the balloon orchid extract was treated and cultured for 12 hours, washed with 1 x PBS, lysed with 1 x reporter lysis buffer, and luciferase assay kit (Promega) and beta The activity of luciferase was measured using a galactosidase assay kit (Promega). That is, the PPAR alpha activity was measured as &quot; luciferase activity / beta -galactose activity &quot;.

Wy-14,643 (Calbiochem), the most potent ligand of PPAR alpha, was used as a positive control in this experiment, and DMSO 0.05% was used as a negative control.

According to the above experiment, C2C12 cells were treated with balloon orchid extract to measure the activity of PPARalpha according to concentration, and the results are shown in Table 6 below. The values of PPAR alpha activity in Table 6 below are percentages relative to the negative control (0.05% DMSO).

Activity of PPAR alpha sample PPAR alpha activity (%) DMSO (0.05%) 100 Wy-14,643 (0.5 [mu] M) 294 Wy-14,643 (1 [mu] M) 892 Balloon orchid extract (0.05%) 459 Balloon orchid extract (0.5%) 901

As shown in Table 6, it was confirmed that the balloon orchid extract showed a very strong PPAR alpha activity when compared with the positive control group.

Example  5: Filaggrin's  Promoting effect of expression

After incubation for 1 day, the cells were harvested, and the cDNAs were synthesized. Then, the Taqman stain solution was used for Real -time PCR was performed. The RNA concentration was normalized to S16 ribosomal RNA.

Effect of expression of pillar green depending on concentration (number of repeats = 3) Additive sample Increase in pillar green compared to control Control group (no addition) 1.00 Balloon orchid extract (5 / / ml) 25.1 Balloon orchid extract (10 ㎍ / ml) 53.6

As shown in Table 7, the balloon orchid extract has the ability to promote pilar green expression of human keratinocyte HaCaT cells.

Example  6: To improve the turn-over of the skin  About Efficacy

About the nutritional cream of the following formulation example 2, the effect of the improvement of the exfoliation of the skin in the healthy twenties to 50 women was tested as follows.

Twenty of 20 women aged 20 to 50 years were given a 1.5% solution of DHA (Dihydroxyacetone, Sigma Aldrich, USA) for 8 hours, followed by deposition. The preparation cream was applied twice daily to the test site. After 4 days of application, 8 days after application, 12 days after application, photographs were taken using instrument Chromameter CR-400 (Minolta, Japan) and skin lightness of DSLR and DSLR. The measured values were evaluated by three mean values excluding the maximum value and the minimum value. The higher the improvement in the skin brightness of the deposition site, the better the improvement of the exfoliation. The results are shown in Table 8 below.

Skin brightness improvement effect division Four days after application Eight days after application After 12 days of application Improvement rate (%) 1.48 5.20 13.94

As shown in Table 8, when the nutritional cream according to the present invention was used, it was found to have the effect of improving the exfoliation of the skin.

Formulation example  1: Manufacture of pharmaceutical preparations

1. Preparation of tablets

Balloon orchid extract 0.2 mg

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

As in the following composition, a nutritional cream containing the balloon orchid extract as an active ingredient was prepared by a conventional method.

Balloon orchid extract 0.2 wt%

Beta-1,3-glucan 5.0 wt%

Wax 10.0 wt%

Polysorbate 60 1.5 wt%

&Lt; tb &gt; &lt; tb &gt; &lt; tb &gt;

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: Preparation of external preparation for skin

1. Manufacture of ointment

An ointment containing the balloon orchid extract as an active ingredient was prepared according to a conventional method, as shown below.

Balloon orchid extract 0.5 wt%

Beta-1,3-glucan 10.0 wt%

Wax 10.0 wt%

Polysorbate 60 5.0 wt%

&Lt; tb &gt; &lt; tb &gt; &lt; tb &gt;

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 (19)

A composition for regenerating skin comprising an orchid extract of balloon as an active ingredient.
The method according to claim 1,
The composition for skin regeneration is an extract obtained by extracting a balloon orchid with one or more selected from the group consisting of water and an organic solvent.
3. The method of claim 2,
The organic solvent may be a polar solvent containing a lower alcohol having 1 to 5 carbon atoms, ethyl acetate, or acetone; A nonpolar solvent comprising ether, chloroform, benzene, hexane or dichlorohexane; Or a mixed solvent thereof.
The method according to claim 1,
Wherein the composition is for the manufacture of a medicament for skin regeneration, a cosmetic preparation or a food product.
A composition for improving skin wrinkles or elasticity, comprising a balloon orchid extract as an active ingredient.
6. The method of claim 5,
Wherein the composition is for the manufacture of a medicament for the improvement of skin wrinkles or elasticity, a cosmetic composition or a food product.
A composition for antioxidation comprising a balloon orchid extract as an active ingredient.
8. The method of claim 7,
Wherein the composition is for the production of antioxidant medicines, cosmetics or foods.
A composition for antiinflammation comprising an orchid extract of balloon as an active ingredient.
10. The method of claim 9,
Wherein the composition is for the manufacture of an anti-inflammatory medicament, a cosmetic or food product.
A composition for improving atopy comprising a balloon orchid extract as an active ingredient.
12. The method of claim 11,
Wherein the composition is for the manufacture of a medicament for the improvement of atopy, a cosmetic preparation or a food product.
A composition for moisturizing the skin, comprising as an active ingredient a balloon orchid extract.
14. The method of claim 13,
Wherein the composition is for skin moisturizing medicines, cosmetics or foods.
A composition for improving skin texture, comprising a balloon orchid extract as an active ingredient.
16. The method of claim 15,
Wherein the composition is for the manufacture of a medicament for the improvement of skin texture, a cosmetic preparation or a food product.
A composition for promoting stem cell activity comprising a balloon orchid extract as an active ingredient.
18. The method of claim 17,
Wherein the composition is for promoting stem cell proliferation or stem cell retention.
18. The method of claim 17,
Wherein the composition is for the production of a cosmetic for promoting stem cell activity.