KR20120135550A - Composition for skin external application comprising culture extract of rosa placenta and functional food comprising the same - Google Patents

Abstract

PURPOSE: An external use skin composition and a functional food containing a rose placenta tissue culture or extract thereof are provided to activate skin cells and to ensure skin regeneration and anti-wrinkling effects. CONSTITUTION: An external use skin composition for improving skin condition contains 0.1-10 wt% of rose placenta tissue culture or extract thereof as an active ingredient. The culture or extract is prepared in a powder form. The composition is used for proliferating and regenerating skin cells and for enhancing antioxidation, collagen synthesis, and anti-aging. A method for manufacturing the composition comprises a step of isolating and culturing rose placenta; and a step of preparing the composition containing the culture or extract. A functional food contains the culture or extract. [Reference numerals] (AA) Cell proliferation effect of rose placenta tissue culture extract on human keratinocytes; (BB) Rose extract; (CC) Rose placenta tissue culture extract; (DD) Proliferation effect(%); (EE) Control group

Description

Composition for Skin External Application Comprising Culture Extract of Rosa Placenta and Functional Food Comprising the Same}

The present invention relates to a composition for external application for skin containing a rose threshing tissue culture or an extract thereof, and a functional food containing the same.

About 300,000 kinds of plants are known around the world, and it is found that the plants contain useful components that exhibit various physiological activities. These useful components are secondary metabolites that are produced secondary to the normal metabolism of cells and accumulate in specific places within the cell. From a biochemical or ecological point of view, they are used to protect the plant itself from microorganisms or external animals, There is a growing interest in that it is an advantageous material. Secondary metabolites produced by plants include alkaloids, flavonoids, carotenoids, glycosides, and terpenoids. These secondary metabolites have many effects such as moisturizing, soothing and converging skin, UV protection, whitening, exfoliation, skin aging prevention, antioxidant, skin wrinkle improvement, antibacterial, anti-inflammatory, anti-cancer.

The bioactive substance, which is a secondary metabolites, is extracted through in-flight cultivation, that is, cultivation aseptically in a nutrient-containing medium. To produce. Through plant tissue culture, the technology for producing a large amount of useful substances by tissue culture, such as a plant, has been developed so that its contents are too low to be extracted directly from the plant. The production of secondary metabolites with excellent physiological activity by overcoming many constraints, such as different productivity or limited plant growth under the influence of climate, season, location, etc. This is possible.

On the other hand, as the industry develops, environmental pollution becomes serious and interests on anti-aging, whitening, etc. of the skin are increasing, and efforts are being made to develop products for sun protection, soothing and converging skin, antioxidant, and wrinkle improvement. In particular, cosmetic preparation using placenta-derived extracts has been attempted, but animal-derived placenta extracts are concerned about pathogenic and allergic problems such as Bovine Spongiform Encephalopathy (BSE) and cholera in pigs. When directly extracted from the mammalian placenta including cosmetic ingredients, according to the extraction method may contain a large amount of impurities not involved in skin care, there is a risk of causing an immune rejection reaction, especially in the case of skin, especially skin wounds, In placenta, for reasons of ethics, The sale of cosmetics is systematically prohibited. On the other hand, the development of the external preparation composition or functional food using the plant pollen extract is insufficient.

The inventors of the present invention, while trying to manufacture a skin external preparation composition or food having a skin improvement ability, isolating the tissues of the tissues of the rose, and cultured through the plant tissue culture, the culture of the tissues of the rose or its extracts are grown and The present invention was completed by activating proliferation and confirming that there are antioxidant and anti-wrinkle effects.

Accordingly, it is an object of the present invention to provide a skin external preparation composition for improving skin, comprising a rose pollen tissue culture or an extract thereof as an active ingredient.

Still another object of the present invention is to provide a functional food containing the rose pollen tissue culture or its extract as an active ingredient.

Still another object of the present invention is to provide a method for preparing a composition for external application for skin containing the rose pollens tissue culture or extract thereof as an active ingredient.

Still another object of the present invention is to provide a method for producing a functional food containing the rose pollen tissue culture or its extract as an active ingredient.

In order to achieve the above object, the present invention provides a skin external composition for improving the skin containing rose rotaceous tissue culture or its extract as an active ingredient.

The present invention also comprises the steps of: (a) separating and cultivating the tissue in the ovary of the rose; And (b) provides a method for producing a skin external composition for skin improvement containing the culture or rose extract of the Taekwondo tissue comprising the step of preparing a composition containing the culture or extract thereof obtained in step (a). do.

The present invention also provides a functional food containing a culture of rose pollen tissue or an extract thereof.

The present invention also provides a composition for antioxidant containing a culture or extract thereof of rose pollen tissue.

The composition for external application of the rose thoracic tissue culture or extract containing the extract according to the present invention contains the rose thoracic tissue culture or the extract having excellent antioxidant activity, thereby activating the skin cells due to the growth or proliferation of the skin cells, thereby regenerating the skin and preventing the antioxidant. And there is an excellent effect to improve the wrinkle improvement effect.

Figure 1 is a graph confirming the cell proliferation effect of rose fetal tissue culture extract according to the present invention in human keratinocytes.
Figure 2 is a graph confirming the antioxidant capacity of rosettes tissue culture extract.
Figure 3 is a graph confirming the collagen enhancement effect of human fibroblasts of rosette tissue culture extract.
Figure 4 is a graph confirming the cytokine expression rate of rosette tissue culture culture extract.

The present invention relates to a skin external preparation composition for improving skin containing the rose thoracic tissue culture or its extract as an active ingredient, a method for producing the same, and a functional food containing the same.

The present invention in one aspect, the present invention relates to a skin external preparation composition for improving skin containing rose rotaceous tissue culture or extract thereof as an active ingredient. Specifically, the rose rotiferous tissue culture or its extract may be contained in an amount of 0.1 to 10% by weight based on the total weight of the composition. This ratio is only a preferred range, for example, in the following examples, the case of 0.1 to 5% by weight was tested, in addition to 10% by weight, or more than 20%, 30% or more, also excellent skin improvement effect, It is apparent to those skilled in the art that it has functions of skin proliferation, regeneration, wrinkle improvement, and antioxidant activity.

In the present invention, using the rosette tissue culture (culture) itself, the culture is used by filtration, the culture can be used by crushing, or the culture to dry the powder, and then dissolved in purified water can be used have.

In the present invention, "extract" refers to an extract obtained by powdering the rose pollen tissue culture extract, or by various conventionally known extraction methods such as hot water extraction, ethanol extraction.

The extraction method in the present invention is not particularly limited, and examples thereof include cold needle extraction, ultrasonic extraction, reflux extraction, hot water extraction, and the like. Here, in the case of ultrasonic extraction, ultrasonic extraction for 60 minutes at 40kHz using an ultrasonic generator (Elma, Transsonic 1040 / H), and stirred for 2 hours at 65 ℃ reaction, and then 8 to 48 hours in a hot water distillation machine Is heated to 80-100 ° C. to obtain a hydrothermal extract.

Or by extraction using water, an organic solvent, or a mixed solvent thereof. The extracted solution can be used directly or can be concentrated and / or dried. When extracted with an organic solvent, methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixed solvent thereof may be used and extracted by room temperature or warming under conditions where the active ingredient of the herbal medicine is not destroyed or minimized. Depending on the organic solvent to be extracted, the degree of extraction and loss of the active ingredient of the drug may vary, so select an appropriate organic solvent.

In the present invention, the extract may be used by concentration or dilution, or distillate of the extract may be used.

In the present invention, "for skin improvement" means skin protection and skin condition improvement, skin whitening, skin aging and wrinkle prevention or improvement, skin protection and alleviation of skin inflammatory response, immune disease improvement ability, or skin barrier function It is a concept that includes improvement, alleviation of skin irritation, skin cell proliferation and regeneration ability, antioxidant activity, and collagen synthesis enhancement ability.

In the present invention, the meaning of "contained as an active ingredient" means that the composition for external application for skin can contain a skin improving effect, for example, can contain and improve the proliferative activity of skin cells. do. In addition, it means that the composition for external application for skin contains an effective amount capable of exhibiting collagen synthesis or MMP-1,9 inhibitory activity related to the anti-wrinkle effect, or an antioxidant effect or a cytokine expression ability.

In the present invention, a rose is a generic name of a plant belonging to the dicotyledonous plant Rosaceae Rosaceae, and is a shrub-like flowering plant. Roses are deciduous trees, sometimes vines, stalks have thorns, leaves are regenerated, flowers are short-lived, or the length of the stem is shorter and shorter upwards, and each flower is almost coplanar. Run side by side on the mountain. Calyx and petals are usually five, but rarely four. There are several surgery, pistil is a lot in the burn (花 床). Burns are fruit, sometimes swollen, berry-shaped, and contain many hard fruits. Today, the rose refers to the natural hybrids and improved species of these wild species. Each species has many individual variations, and is likely to be a hybrid between species. Roses are flowers that have been cultivated for ornamental and fragrance because of their beautiful shape and fragrance. They are cultivated by the improvement of Rosa hybrida Hort. In the West, it was the most beloved ornamental plant from ancient times, and is one of the most popular ornamental plants in the world today. Western Ancient Rosa gallica , R. centifolia , R. damascena and the like. In modern times, hybrid teas were made from Chinese laurels or from the complex crosses of R. odorata and R. gallica , R. damascena and R. moschata in the Mediterranean. Pure sulfur varieties were produced by hybridization of Iranian native R. foetida and Hybrid Tea. The crossover of brier and laurel crossed Hybird Tea to produce a large, multi-flora floribunda.

In the present invention, "rose" is not limited thereto, but Rosa gallica , Rosa centifola , Rosa damascene , Rosa chinensis , Rosa odorata , Rosa moschata , Rosa foetida , Rosa canina , Rosa rugosa, Rosa davurica Pall , Rosa multiflora , Rosa luciae , Rosa wichuraiana , Rosa gigantea , Rosa alba , Rosa primula , Rosa banksiae, Rosa Mundi , Rosa brunonii , Rosa cinnamomea , Rosa glauca , Rosa spinosissma , Rosa foetida bicolor , Rosa foetida persiana , Rosa fedtschenkoana , Rosa mulliganii, Rosa sericana pteracantha , Rosa hugonis , Rosa banksiae lutea , Rosa banksiae , Rosa roxburghii, Rosa hirtula , Rosa aciculaisis nipponensis , Rosa uchiyamana , Rosa jasminoides , Rosa fujisanensis, Rosa laevigata , Rosa cinnamomea , Rosa nitida , Rosa california , Rosa virginiana , and Rosa Rose genus includes palustris .

Plant tissue culture is used to keep live small plant tissues in It is a technique for growing plant cells, organs and plants according to the purpose of culture by aseptically culturing in vitro . Plant tissue culture can be said to use the basic characteristics of the plant. Plant tissue culture technology is based on the use of the totipotency that can regenerate the plant from the somatic cells of the plant, a unique feature of the plant. That is, by culturing single cells (including protoplasts), leaf and root sections of plants in a medium to which specific nutrients and growth regulators are added, the plants can be regenerated from these cells and tissues. The ability of these plants to totipotency is a strategy for surviving from the damage of the environment and herbivores for the fixed life and long-term survival of the plants. Thus, plants can be said to have the ability to regenerate lost organs. Plant tissue cells isolated from the parent plants can be regenerated into complete plants when cultured in an appropriate culture environment, that is, plants with typical properties are not only academically important in embryology but also for practical use of cosmetic raw materials. Is large.

Placenta of the plant, like the placenta derived from an animal, is an important site that manages seed growth, and is the part where the seed is attached in the ovary of the pistil. It is usually located at the edge of the carpel, which forms an ovary, sometimes in the middle vein of the carpel. It may also be formed in the base of the ovary or in the shape of a column standing upright in the ovary from the base.

In addition, the present invention in another aspect, the step of (a) separating and culturing the tissue of the rosette; And (b) a method for producing a skin for the improvement of skin composition containing rose rosette tissue culture or the extract comprising the step of preparing a composition containing rose rosette tissue culture or extract thereof, rosette tissue tissue culture Or it relates to a functional food production method containing the extract.

In the step (a), specifically, an appropriate medium may be selected for culturing rose dorsal tissue. If there is a medium generally used for plant tissue culture in the art, it can be used without limitation. In plants, MS medium and B5 medium are generally used. For example, the composition of MS medium (1 L standard) is NH ₄ NO ₃ 1650 mg, KNO ₃ 1900 mg, CaCl ₂ H ₂ O 440 mg, MgSO _{4 .7H 2 O 370 mg, KH} 2 PO 4 170 mg, KI 0.83 mg, H 3 BO 3 6.2 mg, MnSO 4 .4H 2 O 22.3 mg, ZnSO 4 .7H2O 8.6 mg, Na 2 MoO 4 .2H 2 O _{0.25 mg, CuSO 4, 5H 2} O 0.025 mg, CoCl 2 .6H 2 O 0.025 mg, FeSO 4 .7H 2 O 27.8 mg, Na 2 EDTA.2H 2 O 37.3 mg, Myoinositol 100 mg, Nicotinic acid 0.5 mg, Pyridoxine -HCl 0.5 mg, Thiamine-HCl 0.5 mg, Glycine 2 mg, Sucrose 30000 mg.

In the present invention, in the step (b), the composition containing the rosette tissue culture culture obtained through the culture of step (a) as it is prepared in a suitable form of the external preparation composition or functional food, or the culture It can be obtained in the form of extract through a known extraction method to prepare a suitable external skin composition or functional food. For example, the culture obtained in the step (a) is powdered after hot air drying to evaporate moisture, As needed, it can manufacture by mixing with an appropriate binder.

In the present invention, the binder is when the rose pollen culture or the extract according to the present invention is provided as a food material, for example, when provided in the form of a liquid, powder (powder), granules, tablets, etc. It is an additive added to improve handling convenience and formability. Examples of such binders include water, alcohols, glycerin, thickened polysaccharides, and the like in powders, lactose, starch, dextrin, sucrose, oligosaccharides, cyclodextrins, cellulose derivatives, granules, tablets, and capsules. And the like, lactose, starch, dextrin, sucrose, oligosaccharide, cyclodextrin, cellulose derivative, sucrose ester, fatty acid ester and the like.

In the present invention, the composition for external application for skin may be a cosmetic composition or a pharmaceutical composition.

In the cosmetic composition, an acceptable carrier may be included in the cosmetic formulation. Herein, "acceptable carrier in cosmetic preparation" means a compound or composition already known and used that may be included in a cosmetic preparation or a compound or composition which will be developed in the future, and which has no toxicity, instability or irritation that is acceptable to the human body upon contact with the skin. Say nothing.

The carrier may be included in the skin topical composition of the present invention in an amount of from about 1% by weight to about 99.99% by weight, preferably from about 90% by weight to about 99.99% by weight of the composition, based on the total weight thereof. However, since the ratio varies depending on the formulations described below in which the composition for external application for skin of the present invention is prepared, and its specific application site (face, neck, etc.) or its preferable application amount and the like, And should not be construed as limiting the scope of the invention.

Examples of the carrier include an alcohol, an oil, a surfactant, a fatty acid, a silicone oil, a wetting agent, a moisturizer, a viscous modifier, an emulsifier, a stabilizer, an ultraviolet scattering agent, an ultraviolet absorber, a coloring agent and a perfume. The compounds / compositions which can be used as the above-mentioned alcohol, oil, surfactant, fatty acid, silicone oil, humectant, humectant, viscous modifier, emulsifier, stabilizer, ultraviolet scattering agent, ultraviolet absorber, The person skilled in the art can select and use the appropriate substance / composition.

As an embodiment of the present invention, the external preparation composition for skin according to the present invention may include glycerin, butylene glycol, propylene glycol, rolled polyoxyethylene hydrogenated castor oil, ethanol, triethanolamine, etc. , Preservatives, paints, colorings, purified water and the like may be included as necessary.

The composition for external application for skin according to the present invention can be prepared in various forms such as lotion, essence, gel, emulsion, lotion, cream (underwater type, water type, multiphase), solution, suspension ), Capsules (soft capsules, hard capsules) having a coating such as anhydrous products (oil and glycol), gel, mask, pack, powder or gelatin.

The skin of the present invention includes not only the face but also the scalp and whole body. The composition for external application for skin which can be applied to such scalp includes shampoo, rinse, treatment, hair removal agent, etc., and a body cleanser And the like can be manufactured in various forms.

The method for preparing the composition for external application for skin containing rose thoracic tissue culture extract according to the present invention is not limited to the above-described preparation method, and those skilled in the art to which the present invention pertains may partially modify the preparation method. As an external composition for skin preparations containing rose fetal tissue culture extract according to the present invention can be prepared.

In particular, the external preparation composition for the skin may be prepared in the form of general emulsion formulations and solubilized formulations using a conventionally known production method, in addition to the production method specifically disclosed in the present invention.

When the cosmetic composition is prepared, cosmetics of the emulsion formulations include nutrient cosmetics, creams, essences, etc., and cosmetics of the solubilized formulations include soft cosmetics. In addition, by containing a dermatologically acceptable medium or base, it can be prepared in the form of topical or systemic adjuvants commonly used in the field of dermatology.

In addition, formulations of suitable cosmetics include, for example, emulsions, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes), nonionics obtained by dispersing an oil phase in a solution, gel, solid or pasty anhydrous product, aqueous phase, for example. It may be provided in the form of a vesicle dispersant in the form of a cream, skin, lotion, powder, ointment, spray or concealed stick. It may also be prepared in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

In addition, the topical skin composition of the present invention may further comprise fatty substances, organic solvents, solubilizers, thickeners and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ions Any commonly used in type or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics It may contain adjuvants conventionally used in the cosmetic or dermatology field as other ingredients. In addition, the above components may be introduced in an amount generally used in the dermatology field.

Such a composition for external application for skin according to the present invention has excellent skin cell regeneration and proliferative ability, including improvement of skin condition, antioxidant activity, prevention of skin aging according to collagen synthesis enhancement ability, skin whitening, relief of inflammation by excellent cytokine expression ability, and the like. It includes a form of functional cosmetics that can function in the treatment / prevention of immune diseases.

In the present invention, the pharmaceutical composition expresses various cytokines of VEGF, EGF, FGF, TGF-β1, IL-1α, IL-6, IL-8 and TNF-α shown in the following Examples, In particular, it has an excellent level of cytokine expression of EGF, TGF-β1 and TNF-α, and can function as a pharmaceutical composition for treating and preventing immune diseases.

Such pharmaceutical compositions may comprise "pharmaceutically acceptable carriers", in addition to the active rosette tissue culture or extracts thereof, which carriers may be diluents, lubricants, binders, disintegrants, sweeteners, stabilizers and preservatives. It may be selected from the group consisting of. . The pharmaceutical composition may further comprise an additive. The additives may include flavoring agents, vitamins, and antioxidants. Examples of the diluent include lactose, dextrin, tapioca starch, corn starch, soybean oil, microcrystalline cellulose or mannitol as a carrier, magnesium stearate or talc as a lubricant, , And the binding agent may be polyvinylpyrrolidone or hydroxypropylcellulose. Examples of the disintegrant include carboxymethylcellulose calcium, sodium starch glycolate, polacrilin potassium, or crospovidone; examples of sweeteners include white sugar, fructose, sorbitol, or aspartame; stabilizers include sodium carboxymethylcellulose, Or xanthan gum, and the preservative may be methyl paraoxybenzoate, propyl p-hydroxybenzoate, or potassium sorbate.

The pharmaceutical composition may be formulated in conventional pharmaceutical formulations known in the art. The pharmaceutical compositions may be formulated and administered in the form of oral dosages, injections, suppositories, transdermal dosage forms, and nasal dosages. For example, the formulation may be a formulation for oral administration such as liquids, suspensions, powders, granules, tablets, capsules, pills, or extracts.

In another aspect, the present invention relates to a functional food or food additives comprising the rose rotiferous tissue culture or extract thereof.

Rose tissue of the present invention or the extract itself is almost no toxicity and side effects can be used safely in food.

In the present invention, the term "functional food" is the same term as food for special health use (FOSHU), in addition to the nutritional supply, the processed food, medical effect is effectively processed to appear bioregulatory function It means high food. In the present invention, the term "functional food" is used interchangeably with the terms health food, health supplement food. "Health food (health food)" means a food that has an active health maintenance or promotion effect compared to the general food, health supplement food (health supplement food) means a food for health supplement purposes. These functional foods include tablets, soft capsules, hard capsules, powders, granules, liquids, pills, beverages, jelly, gummi, candy, cookies, biscuits, and energiba to achieve the effect of skin whitening, wrinkle improvement, and antioxidant activity. It can be prepared in various forms such as balanced nutrition foods.

The rose thoracic tissue culture of the present invention or the extract thereof is a skin protection and improvement effect, skin whitening effect, skin irritation, skin aging and wrinkle prevention and improvement effect, skin protection and skin inflammatory response alleviation effect, or skin It may be added to foods or beverages for the purpose of restoring barrier function. In addition, it contains physiologically active substances such as polyphenols and flavonoids, and by providing antioxidant activity, it is possible to restore the body's functions, including metabolic activity of the antioxidant action, and prevent aging. In addition, it expresses various cytokines of VEGF, EGF, FGF, TGF-β1, IL-1α, IL-6, IL-8 and TNF-α, and among them, specifically for EGF, TGF-β1 and TNF-α. It has an excellent level of cytokine expression and can function to improve immune diseases. At this time, the amount of the rosette tissue culture extract in the food or beverage can be generally added in 0.01 to 5% by weight of the total weight of the food, the beverage is in the ratio of 0.01 to 2g, preferably 0.3 to 1g based on 100ml Can be added.

In the present invention, the rose thoracic tissue culture or its extract may be diluted with an inorganic solvent, an organic solvent including purified water, or may be contained in an undiluted state.

There is no particular limitation on the liquid component except that the rose fetal tissue culture of the present invention or the beverage to which the extract is added is an essential ingredient in the indicated ratio and the rose fetal tissue culture or its extract is not limited. Eggplant flavourant or natural carbohydrate, etc. may be contained as additional components. Examples of the natural carbohydrate include monosaccharides such as glucose, fructose, galactose, mannose, and the like; Disaccharides such as maltose and sucrose; Conventional sugars such as dextrin, cyclodextrin, and the like; And sugar alcohols such as xylitol, sorbitol, erythritol, and the like. As said flavoring agent, natural flavoring agents, such as tautin and a stevia extract (for example, rebaudioside A, glycyrrhizin, etc.), and synthetic flavoring agents, such as saccharin and aspartame, can be used advantageously. The ratio of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g, per 100 ml of the composition containing the rose fetal tissue culture extract of the present invention.

In addition to the above, food and beverages include various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic and natural flavors, colorants and flavor enhancers (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and salts thereof, Organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. Others may contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently. The proportion of such additives is generally selected from the range of 0 to about 20% by weight per 100% by weight of the functional food of the present invention.

The rose fetal tissue culture or extract of the present invention may be used directly by adding to food, but the target food is directly immersed in the rose fetal tissue culture or its extract or by spraying the rose fetal tissue culture extract on food to protect the skin and The improvement effect, the skin whitening effect, the skin aging and the prevention and improvement effect of wrinkles, the skin protection, the alleviation effect of the inflammatory response of the skin, and the restoration effect of the skin barrier function can be obtained. In addition, it contains a physiologically active substance such as polyphenols and flavonoids, and by providing antioxidant activity, it is possible to restore the function of the body and prevent aging. In addition, various VEGF, EGF, FGF, TGF-β1, IL-1α, IL-6, IL-8 and TNF-α cytokines are expressed, and among them, specifically, for EGF, TGF-β1 and TNF-α It has an excellent level of cytokine expression and can function to improve immune diseases.

In the present invention, the functional food may be in the form of powders, granules, tablets, soft capsules, hard capsules, liquid, pills, jelly, gummi, candy, cookies, biscuits, balanced nutritious foods or beverages, preparation in these forms Techniques can be prepared using methods known in the art, without being significantly limited, including the following preparations.

Functional foods as described above, it can be made by providing a rosette tissue culture or extract thereof according to the present invention as a food additive.

The food additives include organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, phosphates such as sodium phosphate, potassium phosphate, acid pyrophosphate and polyphosphate (polymeric phosphate), polyphenols, catechin, and α-tocopherol. It may further include any one or more of natural antioxidants, such as, rose mary extract, licorice extract, chitosan, tannic acid, phytic acid.

In addition, the present invention, the food additives in food, fungicides, spices, seasonings, various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic and natural flavors, colorants and flavor enhancers (cheese, chocolate, etc.), Pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, etc. It provides a method of using a food additive. At this time, the food additive may be added to the food by dipping, spraying or mixing the food, the ratio of these additives is not so important, but with respect to 100 parts by weight of the total composition, rose rotaceous tissue culture of the present invention or its extract 0 to It is usually selected in the range of about 20 parts by weight.

The food is natural foods such as cereals, potatoes, beans, vegetables, fruits, seafood, meat, milk, seaweeds; Or processed foods, such as bread, confectionery, ramen, and ice cream; Or drinks.

In another aspect, the present invention relates to a composition for antioxidant containing such a culture or extract, as the culture of the rose pollen tissue or extract thereof has excellent antioxidant capacity. Such an antioxidant composition can promote the function of metabolism, restore the function of the body, and prevent aging. In addition, it expresses various VEGF, EGF, FGF, TGF-β1, IL-1α, IL-6, IL-8 and TNF-α cytokines and expresses high levels of cytokines for EGF, TGF-β1 and TNF-α. Since it has the ability to function to improve immune diseases, such an antioxidant composition may thus include an anti-aging composition and a form for improving immune diseases.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as being limited by these examples.

In particular, in the following examples, but confirmed the efficacy of the rosette left tissue culture culture extract, it will be apparent to those skilled in the art that such an effect even for the non-extract culture.

Roses according to the invention Tissue  Preparation of Culture Extracts

1.1 Rosa Damascena ) Isolation of the tissue

For cultivation, seed seeds and ovaries in rose buds were sterilized for 5 minutes with 70% ethanol and 0.5% sodium hypochlorite and washed several times with sterile water. Carefully peeled the seed barrier using tweezers, separated the urine and incubated for 2 to 3 weeks in MS medium (Duchefa, Cat # M0256) containing 5% Sucrose.

1.2 Cultivation and Mass Production of Separated Rosettes

Aseptic isolated rosettes were passaged at intervals of 2 to 3 weeks in MS medium containing 5% Sucrose. Thereafter, using a balloon-type bioreactor (Samsung Science), the liquid medium of the same composition, except for Agar, was adjusted to a temperature of 25 ° C., and the air supply amount was adjusted to 0.1 vmv at 14 days intervals, followed by incubation and proliferation every 14 days.

1.3 Preparation of Rose Fetal Tissue Culture Extracts

The cultured rose thoracic tissue culture was harvested, dried with clean tissue, and dried in a dryer at 60 ° C. for 2 days. 100 g of dried rosette tissue culture powder was placed in a container, and 10 L of purified water was added thereto, followed by ultrasonic extraction at 40 kHz for 60 minutes using an ultrasonic generator (Elma, Transsonic 1040 / H), followed by 2 hours at 65 ° C. After stirring for a while, the mixture was heated for 8 to 48 hours and heated to 80 to 100 ° C. in a hot water distillation unit to obtain a hot water extract. After extraction, the solids were removed by filtration through a mesh to prepare a rosette tissue culture extract.

Comparative example  1: Preparation of Rose Petal Extract

The rose petals were harvested and dried sufficiently in a drier at 60 ° C. for 2 days. 100 g of dried rose petals were placed in a container, and 10 L of purified water was added thereto, followed by ultrasonic extraction at 40 kHz for 60 minutes using an ultrasonic generator (Elma, Transsonic 1040 / H), followed by stirring at 65 ° C. for 2 hours. After heating, the mixture was heated at 80 to 100 ° C. for 8 to 48 hours in a boiling water distillation unit to obtain a hot water extract. After extraction, the resultant was filtered through a mesh and solids were removed to prepare a rose extract.

Experimental Example  1: rose according to the invention Tissue  Influence test on cell growth and proliferation of culture extract

In order to determine the proliferative activity of the skin cells of the rosette tissue culture culture extract according to the present invention, human keratinocytes (Human Skin Keratinocyte, HaCaT) were cultured by aliquoting from the American Type Culture Collection (ATCC). Cells were seeded in 24 well culture plates at a concentration of 1 × 10 ⁴ cells / ml. Medium was used DMEM (Dubelcco'S Modified Eagle Medium, BRL, USA) containing 10% FBS. After 48 hours of incubation in DMEM containing 10% FBS and incubated by 25 to 30% of the surface of the culture vessel, the rosette tissue culture extract prepared in Example 1 was replaced with FBS-free DMEM containing 0.1% to 5% by weight. And further incubated for 24 hours. After incubation, 50 µl of 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide (MTT, Sigma M5655, USA) solution (2.5 mg / ml) was added and 3 hours After further incubation, the supernatant was removed, 200 μl of Dimethylsulfoxide (DMSO, Sigma D2650, USA) solution was added to each well, followed by stirring for 20 minutes to dissolve the formazan crystals formed. The absorbance was measured at 570 nm with an Enzyme-Linked Immunosorbent Assay (ELISA). The degree of proliferative activity of the skin cells was calculated as a percentage based on the following formula based on the absorbance intensity of the control group using pure water.

[Equation 1]

Cell growth effect (%) = (absorbance at extract treatment / absorbance of control) × 100

Treatment concentration (% by weight) Cell proliferation effect Control group 100 Rose petals
extract 0.1 99 0.5 100 One 102 5 105 rose
Tissue
culture
extract 0.1 102 0.5 109 One 116 5 124

As a result of the effect on the growth and proliferation of the human keratinocytes (keratinocyte), as shown in Table 1 and Figure 1, when the 5% by weight rosette tissue culture extract containing 124% compared to the control group The cell proliferation effect could be confirmed. 1 wt% rose fetal tissue culture extract showed 116% growth compared to the control group, and 0.5 wt% and 0.1 wt% rose fetal tissue culture extract contained 109% and 102% growth compared to the control group. Showed. Therefore, the rose pollen tissue culture extract according to the present invention showed an effect on the skin cell growth and the activation of proliferation, whereas the rose petal extract, which is not the tissue of the left pollinate tissue culture, had insufficient cell proliferation effect.

Experimental Example  2 rose according to the invention Tissue  Of culture extract Antioxidant ability  Efficacy test

Antioxidant activity of the rose thoracic tissue culture extract according to the present invention was used as an Antioxidant Assay Kit (Cayman, Ann Arbor, Mich.) And was tested according to the manufacturer's instructions.

Antioxidant Capacity of Rose's Peripheral Tissue Culture Extract was determined using Antioxidant Assay Kit (Cayman, Cat No. 709001, Antioxidant Kit). It was measured by conversion to Trolox concentration.

Treatment concentration (% by weight) mM Trolox equiv. Negative Control (Water) 0 Rose petals
extract 0.1 0.12 0.5 0.16 One 0.21 5 0.28 rose
Tissue
culture
extract 0.1 0.28 0.5 0.37 One 0.4 5 0.45 Positive control group
100 μM Vitamin C 0.42

As a result, as shown in Table 2 and Figure 2, the concentration of antioxidant rosette doeneun tissue culture extract (Dose-Dependent) increased the antioxidant capacity. The 5 wt% rosette tissue culture extract showed higher antioxidative potency than the positive control 100 μM vitamin C, indicating the high antioxidant capacity of the rosette tissue culture extract. In contrast, the rose petal extract had a very low antioxidant activity as compared to the rosette tissue culture extract.

Experimental Example  3: rose according to the invention Tissue  Wrinkle improvement (collagen synthesis) effect test of culture extract

Human Skin Fibroblasts at 37 ° C., 5% CO ₂ Incubated in Dulbecco's Modified Eagle's Medium, Gibco, USA (DMEM) containing 10% FBS, Penicillin (50 U / ml) and Streptomycin (50 / ml) in a cell incubator with constant humidity, 1 × 10 ⁵ 500 µl aliquots were injected into 24 well plates at dogs / ml and incubated for 24 hours. Insert the medium containing the control sample (DMEM medium) and rose fetal tissue culture extract of Example 1 in various concentrations of 0.1 to 5% by weight, 37 ℃, 5% CO ₂ for 48 hours Lt; / RTI > After 48 hours, the amount of newly synthesized collagen was measured by taking 20 μl of the supernatant of the medium and measuring the resultant using a Procollagen Type I C-Peptide EIA Kit (PICP, Takara, Cat No. MK101). The results of experiments were obtained in the same manner as the above-described method of extracting rose petals of Comparative Examples. The PICP amount was converted into ng / 1 × 10 ⁵ cells.

Treatment concentration (% by weight) Increase in collagen biosynthesis (%) Rose petals
extract 0.1% 100 0.5% 100 One % 106 5% 115 10% 119 rose
Tissue
culture
extract 0.1% 103 0.5% 110 One % 123 5% 139 10% 157 Positive control retinol 25IU  127

The control value was set at 100, and the increase rate of collagen biosynthesis of the rose petal extract treatment group, the rose thoracic tissue culture extract treatment group, and the positive control group was as shown in Table 3 and FIG. 3.

As shown in Table 3 and Figure 3, the composition of the present invention can be seen that the biosynthesis amount of collagen increases as the content of the rose cadaver tissue culture extract is increased, the composition of the present invention is superior to the composition of the present invention as compared to the rose extract It can be seen that the effect.

Experimental Example  4: rose according to the invention Tissue  Of culture extract Cytokine  Expression assay

In order to determine the expression rate of cytokine in the rosette tissue culture extract according to the present invention, Fibroblast (Cell Lines Service, Germany), a human fibroblast, was inoculated in 24 well culture plates at a concentration of 1 × 10 ⁵ cells / ml. . When cultured in DMEM (Dubelcco'S Modified Eagle Medium, BRL, USA) containing 10% FBS and incubated by 25-30% of the surface of the culture vessel, FBS- containing 1% of the rose fetal tissue culture extract prepared in Example 1 Free DMEM was replaced with FBS-free DMEM used as a control, and further incubated for 48 hours. The expression rates of cytokines of VEGF, EGF, FGF, TGF-β1, IL-1α, IL-6, IL-8 and TNF-α were investigated using HUMAN CYTOKINE ASSAY KIT (Millipore). Experimental method was carried out according to the instructions of HUMAN CYTOKINE ASSAY KIT.

Cytokine Types pg / ml Control group VEGF 19 EGF 16 FGF 14 TGF-β1 21 IL-1? 20 IL-6 21 IL-8 12 TNF-α 21 1 wt% rose fetal tissue culture extract VEGF 24 EGF 36 FGF 19 TGF-β1 52 IL-1? 24 IL-6 23 IL-8 24 TNF-α 50

As shown in Table 4 and Figure 4, the expression rate of the cytokine is expressed in significant levels of EGF, TGF-β1 and TNF-α when treated with 1% rosette tissue culture extract compared to the untreated control group I could confirm it.

These results indicate that the rose pollen tissue culture extract of the present invention has a cytokine expression effect, and thus can function as an immunomodulator.

Experimental Example  5: rose according to the invention Tissue  Bioactive substance content analysis experiment of culture extract

(1) Total polyphenol content was measured by Folins-Denis method which is widely used as an analytical method. 0.5 ml of Folin reagent was added to 0.5 ml of rosette tissue culture extract and allowed to stand for 3 minutes, and then 0.5 ml of 10% Na ₂ CO ₃ solution was added thereto. After the mixture was left for 1 hour, the absorbance was measured at 760 nm using a spectrophotometer (UV / VIS spectrometer, Jasco, Japan). The total polyphenol content of the sample was expressed as mM gallic acid equivalent by preparing a standard curve with gallic acid.

Treatment concentration (% by weight) mM gallic acid equiv. Control (water) 0.05 Rose petals
extract 0.1 0.46 0.5 0.60 One 8.79 5 10.94 rose
Tissue
culture
extract 0.1 4.31 0.5 30.3 One 38.0 5 38.7

As shown in Table 5, it was confirmed that the total concentration of polyphenols increased in a concentration-dependent manner compared with the rose petal culture extract.

(2) Total flavonoid content was measured according to the method of Moreno et al. After adding 0.1 mL of 10% aluminum nitrate to 0.5 mL of rosette tissue culture extract, 0.1 mL of 1 M potassium acetate and 4.3 mL of ethanol were mixed in this order, and the absorbance was measured at 415 nm by reacting for 40 minutes at room temperature. Using a standard curve with catechin as a standard, it was calculated as the content per gram of rose pollen tissue culture extract.

Treatment concentration (% by weight) mg / g catechin equiv. Control (water) 1.02 Rose petals
extract 0.1 2.17 0.5 2.30 One 2.54 5 3.77 rose
Tissue
culture
extract 0.1 5.5 0.5 7.74 One 7.94 5 8.78

As shown in Table 6, it was confirmed that the total concentration of flavonoids increased in a concentration-dependent manner compared to the rose petal extract of rose cadaver tissue.

Such polyphenols and flavonoids have an antioxidant effect, and as a substance to prevent aging, rose fetal tissue culture extract of the present invention contains such polyphenols and flavonoids, and thus, functions to enhance the function of the living body and activate the body. It was confirmed that there is.

Preparation of Cosmetic Composition

As cosmetics containing the rose locust tissue culture extract of the present invention as an active ingredient, cosmetics of emulsified formulations such as nutrient cosmetics, creams and essences, and cosmetics of solubilized formulations such as softening cosmetics were prepared.

Manufacturing example  2-1: lotion

It was prepared according to a conventional lotion preparation method according to the following prescription.

Raw material name Weight% (w / w) glycerin 5.0 Dipropylene glycol 3.0 Hyaluronic acid 0.5 Polyoxyethylene Cured Castor Oil 0.1 Polyethylene oleyl ethyl 0.1 ethanol 5.0 antiseptic 0.15 Beauty Suitable amount flash Suitable amount Rose Teal Tissue Culture Extract 2.0 Purified water to 100

Manufacturing example  2-2: essence

It was prepared according to a conventional essence preparation method according to the following prescription.

Raw material name Weight% (w / w) Cetostearyl alcohol 1.0 Self-emulsifying monostearic acid 1.0 Wax 0.5 Squalane 5.0 Isocetyloctanoate 3.0 Dimethylsiloxane 0.3 Sorbitan monosulfate 0.5 Polyethylene glycol monostearate 8.0 glycerin 4.0 Propylene glycol 0.2 Carboxy polymer 0.22 Triethanolamine 0.25 antiseptic Suitable amount Spices Suitable amount flash Suitable amount Rose Teal Tissue Culture Extract 7.0 Purified water to 100

Manufacturing example  2-3: lotion

It was prepared according to the conventional lotion preparation method according to the following prescription.

Raw material name Weight% (w / w) Cetostearyl alcohol 0.8 Self-emulsifying monostearic acid 1.0 Wax 0.5 Stearic acid 0.5 Liquid paraffin 7.0 Squalane 5.0 Macadamia oil 3.0 Isocetyloctanoate 2.0 Dimethylsiloxane 0.3 Sorbitan monosulfate 0.5 Polyethylene glycol monostearate 1.2 glycerin 4.0 Propylene glycol 4.0 Betaine 4.0 Carboxy polymer 0.12 Triethanolamine 0.15 antiseptic 0.25 Spices Suitable amount flash Suitable amount Rose Throne Tissue Extract 5.0 Purified water to 100

Manufacturing example  2-4: cream

It was prepared according to the conventional cream preparation method according to the following prescription.

Raw material name Weight% (w / w) Cetostearyl alcohol 3.0 Self-emulsifying monostearic acid 1.5 Lipophilic monostearic acid 1.5 Wax 0.5 Liquid paraffin 8.0 Squalane 7.0 Isocetyloctanoate 4.0 Refined jojoba oil 4.0 Dimethylsiloxane 0.3 Sorbitan monosulfate 1.0 Polyethylene glycol monostearate 1.2 glycerin 6.0 Propylene glycol 4.0 Betaine 4.0 Xanthan gum 0.06 Triethanolamine 0.10 antiseptic 0.25 Spices Suitable amount flash Suitable amount Rose Throne Tissue Extract 7.0 Purified water to 100

Manufacturing example  2-5: gel

It was prepared according to the conventional gel preparation method according to the following prescription.

Raw material name Weight% (w / w) glycerin 4.0 Propylene glycol 4.0 ethanol 10 Polyoxyethylene Cured Castor Oil 0.1 Carboxy polymer 0.30 Triethanolamine 0.30 antiseptic Suitable amount Spices Suitable amount flash Suitable amount Rose Throne Tissue Extract 1.0 Purified water to 100

Preparation of Functional Foods

Foods containing the rose fetal tissue culture extract of the present invention were prepared as follows.

Manufacturing example  3-1: Wheat Flour Food

0.5 to 5.0 parts by weight of the rose perilla tissue culture extract of the present invention was added to the flour, and bread, cake, cookies, crackers and noodles were prepared using the mixture.

Manufacturing example  3-2: soup  And juicy ( gravies )

0.1 to 5.0 parts by weight of the rose thalca tissue culture extract of the present invention was added to soups and broths to prepare meat products for health promotion, soups and noodles for noodles.

Manufacturing example  3-3: Ground Beef ground beef )

10 parts by weight of the rose perilla tissue culture extract of the present invention was added to the ground beef to prepare a ground beef for health promotion.

Manufacturing example  3-4: Dairy Products dairy products )

5 to 10 parts by weight of the rose perilla tissue culture extract of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

Manufacturing example  3-5: grain powder food ( Wire Manufacturing

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh. Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer. The rose pollens tissue culture extract of the present invention was concentrated under reduced pressure in a vacuum concentrator, and the dried product obtained by drying with a spray and a hot air dryer was pulverized with a particle size of 60 mesh to obtain a dry powder.

The grains, seeds, and rose pollen tissue culture extracts prepared above were formulated in the following ratios.

(30 parts by weight of brown rice, 15 parts by weight of yulmu, 20 parts by weight of barley)

Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds)

Rose thalca tissue culture extract (3 parts)

An appropriate amount of water, etc. may be added to the grain powder food (wire) obtained by the said manufacturing method, and it may eat.

Manufacturing example  3-6: drink

6-1) Manufacture of Health Beverage

Homogeneous blend of subsidiary materials such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%), water (75%), and 5 g of rosette tissue culture extract of the present invention After sterilization, it was prepared by packing it in a small packaging container such as a glass bottle or a plastic bottle.

6-2) Preparation of Vegetable Juice

Vegetable juice was prepared by adding 5 g of the rosette tissue culture extract of the present invention to 1,000 ml of tomato or carrot juice.

6-3) Preparation of Fruit Juice

Fruit juice was prepared by adding 1 g of the rosette tissue culture extract of the present invention to 1,000 ml of apple or grape juice.

Manufacturing example  3-7: Soft Capsule

Safflower oil, corn oil, cottonseed oil, peanut oil, rapeseed oil, sesame oil, rice germ oil, soybean oil, sunflower oil, evening primrose oil, avocado oil, linseed oil, perilla oil, etc. The mixture was mixed with vitamins, minerals, and various health functional materials appropriately to suit the intended function, and adjusted to a slurry state containing emulsifiers such as beeswax and glycerin fatty acid esters. 280 mg of the slurry was used as the contents, and gelatin contained an appropriate amount of glycerin, and was encapsulated with a film or a vegetable film composed of processed starch, glycerin, and thickened polysaccharides, and a total amount of 450 mg soft capsule was thermally formed.

Manufacturing example  3-8: hard  capsule

40 mg of dextrin was mixed with 180 mg of rosette tissue culture extract of the present invention, and 220 mg of the powder was used as the contents, and placed in a hard capsule made of gelatin or cellulose.

As described above in detail a specific part of the content of the present invention, for those skilled in the art, such a specific description is only a preferred embodiment, which is not limited by the scope of the present invention Will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (20)

A composition for external application for skin improvement comprising a culture of rose threshing tissue or an extract thereof as an active ingredient. The composition for applying the external of the skin according to claim 1, wherein the culture of the rose pollen tissue or extract thereof contains 0.1 to 10% by weight based on the total weight of the composition. The composition for applying the external of the skin according to claim 1, wherein the culture of the rose pollen tissue or extract thereof is powdered and contained in the composition. The skin external composition for skin improvement according to claim 1, wherein the external skin composition is skin cell proliferation and regeneration. The external preparation composition for skin according to claim 1, wherein the external preparation composition for skin has antioxidant capacity. The external preparation composition for skin according to claim 1, wherein the external preparation composition for skin has collagen synthesis enhancing ability. The external preparation composition for skin according to claim 1, wherein the external preparation composition for skin aging is used. A method for preparing a skin external composition for skin improvement comprising a culture of rose pollen tissue or an extract thereof comprising the following steps:
(a) separating and incubating the tissues in the ovary of the rosette; And
(b) preparing a composition containing the culture or extract thereof obtained in step (a). The method of claim 8, wherein the step (b) comprises the step of preparing a composition by drying, powdering and mixing the rosette tissue culture obtained in step (a) with purified water. The method of claim 8, wherein step (b) comprises drying the rosette tissue culture obtained in step (a), powdering and mixing the mixture with purified water, and then preparing a composition by ultrasonic extraction or hot water extraction. Characterized in that. Functional food containing the culture of rose thalca tissue or its extract. The functional food according to claim 11, wherein the functional food is for skin improvement. The functional food according to claim 12, wherein the functional food has skin cell proliferation and regeneration ability. 12. The functional food according to claim 11, wherein the functional food has an antioxidant activity. The functional food according to claim 11, wherein the functional food has a collagen synthesis enhancing ability. 12. The functional food according to claim 11, wherein the functional food is for preventing skin aging. The functional food according to claim 11, wherein the functional food has an ability to express EGF, TGF-β1 and TNF-α, thereby improving immune disease. The functional food according to claim 11, wherein the functional food is in the form of powder, granule, tablet, soft capsule, hard capsule, liquid, pill, jelly, gummi, candy, cookie, biscuit, balanced nutrition food or beverage. 19. The functional food according to claim 18, wherein the functional food is prepared in capsule form by diluting or diluting the culture of the rose pollen tissue or its extract. Antioxidant composition containing the culture of the rose thoracic tissue or extract thereof.

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