KR20170025375A - Composition for improving skin - Google Patents

Abstract

The present invention relates to a composition for improving skin. The thistle extract according to the present invention can be used for skin regeneration, wrinkle improvement, skin whitening, anti-aging, anti-aging, anti- Antioxidants and skin-tone improving drugs, cosmetics or foods. 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, skin whitening, antioxidation, 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, it is the desire of everyone to have a whiter skin. It is genetically determined by the concentration and distribution of melanin in human skin, but is also influenced by environmental or physiological conditions such as sunlight, fatigue, and stress. Melanin is produced by a nonenzymatic oxidation reaction after tyrosine, an amino acid, is converted to DOPA or dopaquinone by an enzyme called tyrosinase. Thus, the pathway through which melanin is made is known, but the mechanism by which melanin synthesis, the previous step in which tyrosinase acts, is not yet elucidated.

On the other hand, commonly known whitening ingredients include substances inhibiting the activity of tyrosinase enzymes such as kojic acid and arbutin, hydroquinone, vitamin C (L-Ascorbic acid) There are plant extracts. By inhibiting the synthesis of melanin pigment, they can brighten the skin tone to realize skin whitening, and it is possible to improve skin hypercholesterolemia such as stain or freckles due to ultraviolet rays, hormones or heredity. However, when applied to skin, there is a problem that the use amount is limited due to safety problems such as irritation and redness, or the effect is insignificant so that a substantial effect can not be expected.

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 the star formation is evident in the polygon and star formation that the lines meet [Non-Patent Document 2].

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 Literature 3,4]. 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 5]. 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 6].

In addition, the present invention relates to a composition for skin regeneration, wrinkle improvement, skin whitening, antioxidant and antioxidant, which is more safe and effective in living body, stable in effectiveness, and more effective than substances having existing skin regeneration, wrinkle improvement, skin whitening, The development of a component having an improving activity is strongly desired.

On the other hand, thistle is a perennial herbaceous perennial plant, which is distributed in Korea, Japan, Northeastern part of China, and Wonsi. The scientific name is [Cirsium japonicum var. ussuriense Kitamura], also known as spiny herbs. It is 50 ~ 100㎝ in height. It has white hairs on the whole and spiderlike hair.

Mature plants are called herbaceous plants, and mature roots are herbaceous plants. Hematocrit, hematuria, hepatitis, hepatitis and hypertension. Thistle leaves and flowers are classified into apigenin, luteolin, myristin, camperol, fektorinarin, 5,7-dihydroxy-6,4'-dimethoxy flavone, hepsidulin-7-neo- Anticancer, anti-mutant, antifungal, neuroprotective, and immunity-enhancing activity.

The inventors of the present invention have found that the skin improvement effect of the thistle extract is excellent as a result of intensive efforts to extract a useful substance having skin rejuvenation, wrinkle improvement, antioxidation, whitening and skin texture improving effect in natural products, and completed the present invention .

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. Journal of the European Academy of Dermatology and Venereology. 12: 103-114, 1999 3. The British journal of dermatology. 110: 129-138, 1984 4. Skin research and technology.5: 189-194, 1999 5. Epidermal Stem Cells of the Skin, Cedric Blanpain, Elaine Fuchs, Annual Review of Cell and Developmental Biology, November 2006, Vol. 22, Pages 339-373 6. 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 thistle extract promotes the proliferation of skin stem cells, promotes skin regeneration, promotes collagen synthesis of fibroblasts of skin to improve wrinkles, suppresses melanin synthesis, exhibits a whitening effect, eliminates free radicals The present invention has been accomplished by confirming the effect of promoting stem cell activity by exhibiting an antioxidative effect and promoting proliferation of skin stem cells and exhibiting stem cell maintenance effect in addition to skin texture improving effect due to improvement in skin brightness.

Accordingly, an object of the present invention is to provide a composition for improving skin regeneration, wrinkle improvement, skin whitening, antioxidation and skin texture improvement, which comprises thistle extract as an active ingredient.

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

As a means for solving the above problems, the present invention provides a composition for skin regeneration, wrinkle improvement, skin whitening, antioxidation and skin texture improvement comprising thistle extract as an active ingredient for the manufacture of medicines, cosmetics or foods.

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

The thistle extract according to the present invention promotes skin regeneration by promoting proliferation of skin stem cells, promotes collagen synthesis of fibroblasts of the skin to improve wrinkles, suppresses melanin synthesis, exhibits a whitening effect, And exhibits an antioxidative effect, and exhibits an effect of improving the skin texture due to the improvement of skin brightness, so that it can be used for the manufacture of medicines, cosmetics or foods.

In addition, the thistle extract promotes the proliferation of dermal stem cells and exhibits a stem cell-retaining 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 the skin regeneration, wrinkle improvement, skin whitening, antioxidant and skin texture improvement effect are applied to actual skin, it exhibits highly active skin regeneration, wrinkle improvement, skin whitening, antioxidation, Is low in volatility so as to be able to stay for a sufficient time to show skin regeneration, wrinkle improvement, skin whitening, antioxidation and skin texture improving effect, and maintains the active ingredient stably on the composition or skin , Medicines, cosmetics, food or the like, 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, skin whitening, antioxidant and skin tone improving ingredients are excellent in skin regeneration, wrinkle improvement, skin whitening, antioxidation and skin texture improving activity even at low concentration in vitro, It is difficult to apply to real skin because of its ability to absorb. Other active ingredients have low hydrophilicity, making them difficult to formulate into medicines, cosmetics, and foods. In addition, some skin regeneration, wrinkle improvement, skin whitening, antioxidant and skin tone improvement components may be decomposed or transformed into other compounds when they are exposed to heat, light, or oxygen, There is also.

As can be seen from the following examples, the thistle extract has remarkably excellent proliferation promoting effect on skin stem cells, promoting collagen synthesis, inhibiting melanin formation, antioxidative effect and skin brightness at a low concentration, , Skin whitening, antioxidation and improvement of skin texture, cosmetic composition or food composition.

Accordingly, the present invention provides a composition for improving skin regeneration, wrinkle improvement, skin whitening, antioxidation, and skin texture improvement containing thistle extract as an active ingredient.

In the present invention, thistle is a perennial herbaceous plant of the dicotyledonous plant L. lilium, and its scientific name is [Cirsium japonicum var. ussuriense Kitamura], also known as spiny herb.

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

The thistle extract may be an extract of flowers, leaves, branches, fruits, roots, stems, seeds or mixtures thereof of thistle.

Preferably, the thistle extract may be an extract obtained by extracting thistle with water and / or an organic solvent. The organic solvent may be a polar organic solvent, a nonpolar 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 thistle 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 thistle 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, or hexane. 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, skin whitening, antioxidant and skin tone improving ingredients. Wrinkle improvement, skin whitening, antioxidation and skin texture improving effect of the composition of the present invention can be further enhanced when the composition contains additional skin regeneration, wrinkle improvement, skin whitening, antioxidation and skin texture improving ingredient. 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 ingredients may be included in an amount ranging from 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 and ease of formulation of the thistle 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 " 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 '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.

In the present invention, the term " skin texture improving effect " 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 of an extract capable of promoting regeneration of damaged skin, improving wrinkles, exhibiting a whitening effect, inhibiting or alleviating oxidation of cells, or improving skin texture it means. When the composition of the present invention contains an effective amount of the thistle extract, the skin regeneration effect, the wrinkle improvement effect, the skin whitening effect, the antioxidation effect, and the skin texture improvement effect can be provided. An effective amount of the thistle 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, the thistle extract may be contained at a higher concentration than in the case where it is commercialized as cosmetics that are routinely applied to the skin. Accordingly, the daily dose 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 thistle extract, 1 to 6 times a day ≪ / RTI >

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 thistle extract 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, thickening and gelling agents, softening agents, antioxidants, suspending agents, stabilizing agents, foaming agents, , Ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, barrier agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or external preparations for skin ≪ / RTI > any other ingredients that are commonly used in the skin sciences. The components can also be introduced in amounts commonly used in the field of dermatology.

When the thistle 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.

Furthermore, the composition for skin regeneration, wrinkle improvement, skin whitening, antioxidation 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.

In addition, the cosmetic composition may further contain, in addition to the thistle extract, 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, a fragrance, Or any other conventionally used in cosmetics such as nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, barrier agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, ≪ RTI ID = 0.0 > ingredients. ≪ / RTI >

The cosmetic formulation may contain a relatively high concentration of the thistle extract in the case of a wash-off type cosmetic such as a make-up remover, a cleanser, etc. in which the active ingredient remains on the skin in a short period of time. On the other hand, in the case of leave-on type cosmetics such as cosmetics, milky lotions, creams, essences and the like in which the active ingredient remains on the skin for a long period of time, the lower thistle extract It will be acceptable. In one embodiment of the present invention, though not limited thereto, the composition may contain the thistle extract in an amount of 0.0001 wt% to 10 wt% (preferably 0.0001 wt% to 1 wt%) based on the total weight of the composition . When the composition of the present invention contains less than 0.0001% by weight of the thistle extract, sufficient skin regeneration, wrinkle improvement, skin whitening, antioxidation and skin texture improvement effects can not be expected. When the composition contains more than 10% To prevent unwanted reactions or skin safety problems.

Further, the composition for skin regeneration, wrinkle improvement, skin whitening, antioxidation and skin texture improvement of the present invention can be used for manufacturing food formulations.

The food preparation refers to a food prepared by adding the thistle extract to a food material such as a beverage, a tea, a spice, a gum, or a confection, or encapsulating, pulverizing, or suspending.

Since the food preparation can be routinely ingested, it is very useful because high skin regeneration, wrinkle improvement, skin whitening, antioxidation and skin texture improving effect can be expected.

When the thistle extract is used as a food additive, the thistle extract can be directly added or used together with other food or food ingredients, and can 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 thistle 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 cosmetic formulations including thistle 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  1: Preparation of thistle extract

The thistle root was washed and pulverized, and then cooled in a 10-fold volume of ethanol for 5 days. 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.), and the handle 5 ㎍ / ml thistle extract 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 skin stem cell of the thistle 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 Thistle extract 5 ug / ml 139% Control group - 100%

As shown in Table 1, the skin stem cells in the medium treated with thistle 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) Thistle extract 5 / / mL 2.14 Control group - 1.0

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

Example  1: Promoting collagen synthesis

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

The thistle extract was added to the culture medium of human fibroblasts (7 × 10 ⁴ cells / cm ² ) together with vitamin C and the control (no addition) to a final concentration of 10 μg / mL and 20 μg / After culturing, the culture 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 Thistle extract 10 [mu] g / mL 30.1% Thistle extract 20 / / mL 59.5% Vitamin C 50 / / mL 32.2%

As shown in Table 3, the thistle extract has excellent collagen synthesis ability against human-derived fibroblasts and can obtain a better collagen synthesis effect at a lower concentration than that of vitamin C, which is generally known to have ability to synthesize collagen .

Example 2: Whitening effect - Confirmation of melanin formation inhibitory effect

The thistle extract was added to the culture medium of rat B-16 mouse melanoma cells to examine the whitening effect at the cellular level (Lotan R., Lotan D. Cancer Res. 40: 3345-3350, 1980). At this time, the toxicity of the melanoma cells of the rats before the experiment was evaluated, and the whitening evaluation was performed by selecting the concentration without toxicity.

The thistle extract was added to the medium to a final concentration of 10 μg / mL and 20 μg / mL, and the control group, arbutin, was added to the medium to a concentration of 200 μg / mL and treated with B-16 melanoma cells for 3 days Respectively.

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 measure the amount of melanin produced.

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

Inhibitory effect of melanin formation at the cellular level (number of repeats = 3) sample Inhibition rate (%) Control group (no addition) - Control group 1: Arbutin (200 쨉 g / mL) 29.4 Thistle extract (20 / / mL) 48.6 Thistle extract (10 / / mL) 33.2

As shown in Table 4, the thistle extract showed remarkably superior melanin production inhibitory effect on the melanoma cells of the rat cultured as compared with the known whitening substance arbutin.

Example  3: Antioxidant effect - Free radical scavenging rate

Free radical scavenging activity was measured to determine the antioxidative activity of thistle 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. Samples were prepared at concentrations of 50 ㎍ / mL, 25 ㎍ / mL, 12.5 ㎍ / mL, 6.25 ㎍ / mL and 3.125 ㎍ / mL, respectively, by adding ethanol to the antioxidant, ascorbic acid and reference material, respectively. 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. 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 free radical scavenging activity as the concentration of ascorbic acid and thistle extract required to remove 50% of the free radicals of the control group without additives.

Free radical scavenging rate (IC ₅₀ ) sample Free radical scavenging rate (占 퐂 / mL) Ascorbic acid 5.1 Thistle extract 5.3

As shown in Table 5, the thistle extract showed high antioxidative effect as compared with ascorbic acid, which is a known antioxidant.

Example  4: Exfoliation (turn-over) 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 6 below.

Skin brightness improvement effect division Four days after application Eight days after application After 12 days of application Improvement rate (%) 1.85 5.83 13.25

As shown in Table 6, 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

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

Thistle extract 0.2 wt%

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: Preparation of external preparation for skin

1. Manufacture of ointment

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

Thistle extract 0.5 wt%

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 (15)

A composition for regenerating skin comprising an extract of Thistle as an active ingredient.
The method according to claim 1,
Wherein the thistle extract is an extract obtained by extracting thistle with at least one 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 containing thistle extract as an active ingredient.
6. The method of claim 5,
Wherein the composition is for the manufacture of a medicinal composition for improving skin wrinkles, a cosmetic composition or a food product.
A composition for skin whitening comprising a thistle extract as an active ingredient.
8. The method of claim 7,
Wherein the composition is for skin whitening medicines, cosmetics or foods.
An antioxidant composition comprising a thistle extract as an active ingredient.
10. The method of claim 9,
Wherein the composition is for the production of antioxidant medicines, cosmetics or foods.
A composition for improving skin texture, comprising a thistle 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 skin texture, a cosmetic preparation or a food product.
A composition for stimulating stem cell activity, comprising a thistle extract as an active ingredient.
14. The method of claim 13,
Wherein the composition is for promoting stem cell proliferation or stem cell retention.
14. The method of claim 13,
Wherein the composition is for the production of a cosmetic for promoting stem cell activity.