KR20150130006A - Anti-aging and Skin- Astriction composition for skin external application comprising Aster sphathulifolius Maxim. Cell Culture Extract and Methods for preparing the Same - Google Patents

Abstract

The present invention relates to a composition for external application to the skin with astringency and anti-aging effects, containing a plant cell culture extract from a native plant of Ulleungdo Aster spathulifolius Maxim., and a method for manufacturing the same and, more specifically, to a composition for external application to the skin with astringency and anti-aging effects, containing a plant cell culture, an adventitious root culture, or an extract thereof derived from a native plant of Ulleungdo Aster spathulifolius Maxim., and a method for manufacturing the composition. The composition for external application to the skin containing the plant cell culture, the adventitious root culture, or the extract thereof derived from Aster spathulifolius Maxim. is nontoxic to the skin and excellent in astringing the skin, protecting or strengthening a skin barrier, and synthesizing skin collagen, thereby having effects on improvement of skin wrinkles or elasticity, anti-inflammation, wound healing and the like.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for external application for skin, which has a converging effect and an anti-aging effect containing a culture extract of a Korean native plant cell which is a native plant of Ulleungdo Island, and a preparation method thereof. Cell Culture Extract and Methods for Preparing the Same}

The present invention relates to a composition for external application for skin having convergence effect and anti-aging effect containing a culture extract of a Korean native plant cell, Ulleungdo, and to a method for producing the same. More particularly, the present invention relates to a plant cell culture derived from a Korean native plant Water, an adventitious root culture or an extract thereof, and a method for preparing the same.

As human skin over time, the hormones that regulate metabolism internally

Secretion is decreased, the function of immune cells and the activity of cells are lowered,

Endogenous aging resulting from reduced biosynthesis of reverse protein and biologic proteins

the skin is thinned (in the case of endogenous aging) due to intrinsic aging and photoaging due to various pollutants and ultraviolet ray externally, and the elasticity is decreased. As the skin is exposed to ultraviolet irradiation, And melanin cells are damaged by UVB. Skin aging is divided into chronologic aging, which occurs in areas that are not exposed to sunlight, and actinic aging, which is a combination of chronologic aging and degenerative changes in the exposed parts of sunlight. In chronological aging, characteristic clinical signs of skin are fine wrinkles, atrophy of dermis, and decrease of subcutaneous fat layer. In the photoaging process, coatse wrinkling and furrowing appear and abnormal elasticity material accumulates and the skin thickens and loosens like leather. The phenomenon seen in chronic sunlight-damaged skin is an abnormal increase in the elastic elastosis and proteoglycan of the upper collagen of the dermis and a significant decrease in collagen, the main protein of the dermis. In general, the dermis consists of the majority of type I collagen, some type II collagen, elastin, proteoglycan, and fibronectin.

Collagen gives strength and tension to the skin, which causes external stimulation or force

It protects the skin and it occupies 90% of the dermis layer,

And aging.

Cells constituting the skin are deteriorated in cell activity due to endogenous aging or photoaging,

As the signal transduction system becomes incomplete, MMPs (matrix

metalloproteinases) are increased, collagen biosynthesis is reduced, and wrinkles

It is known that melanin production, which causes skin elasticity and skin darkness, is increased. Therefore, it can inhibit the biosynthesis of MMPs that decompose collagen, which is a substrate substance constituting the skin, substances that can thicken the skin by proliferating skin cells or increasing the matrix substances constituting the skin, inhibiting melanin biosynthesis And the like are found to alleviate skin symptoms such as wrinkles, loss of elasticity, and skin blackening.

The skin barrier (Stratum corneum, Skin barrier) consists of dead keratinocyte (intercellular lipid) and intercellular lipids, protects the skin from external stimuli and protects the skin from moisture evaporation. It is responsible for core functions. In other words, it prevents excessive release of water in the body and prevents harmful substances such as chemicals and microorganisms from entering our bodies. It plays an important role in intercellular lipid stability in keratinocyte envelope, which constitutes the surface of dead keratinocytes. The keratinocyte envelope is composed of protein membranes such as involucrin, loricrin, and filaggrin.

The keratinocytes make the skin barrier through the process of differentiation and currency keratinization. Skin barrier function can be aged or destroyed by external factors, and aged skin is easily destroyed by acetone or tape stripping. Damage to the skin barrier can cause skin moisture loss and wrinkles.

In addition, tight junctions consist of one type of cell junction, claudin, tricellulin, junctional adhesion molecule (JAM) and zona occluden (ZO). The tight junction can be divided into a part contained in the cell membrane and an intracellular part. Occludin and claudin are closely related constitutive proteins contained in cell membranes. Occludin is the first transmembrane protein found in adherent synapses and has four transmembrane domains. Occludin is known to regulate the function of tight junctions as well as constituents of tight junctions. Claudin, a close-coupled conformational protein with Occludin, also has four transmembrane domains. The tight junction is a cell-to-cell linkage that fills the gap between neighboring cells, controls the movement of small molecules, controls the permeation of cells between cells, and maintains the polarity of the cells. Recently, it has been reported that tight junctions play an important role in skin barrier function.

Native plants are plants that spontaneously grow in nature. Ulleungdo has a lot of native plants in particular. Among them, Aster sphathulifolius Maxim.) is a perennial herbaceous plant that grows on the west coast of southern central and southern Korea, along the coast of the East Coast, and is one of the most prevalent species on Ulleungdo. Flowers start to bloom in July and continue to bloom until November. The stem is slightly hard like a tree, the branch is divided a lot, it grows diagonally, height is 30 ~ 60cm, the leaf has many villi on both sides and it is misaligned. Even in winter, the leaves of the upper part remain semi-evergreen, without any damage.

In recent years, many studies have been made on the development of functional foods using plant resources and utilization of them as cosmetic materials, but there are prior patented technologies (Patent Document 1) in which a part of adult extract is used as an antihistamine composition, The research as a result is insignificant. It is also urgent to develop various cosmetic materials for preventing and strengthening skin barrier which plays a key role in skin health.

Patent Document 1: Korean Patent Application No. 10-2010-000128143

The present inventors have found that it is possible to improve the firmness of the skin and to prevent or reduce the visible symptoms of aging such as wrinkles derived from external or internal causes such as sun exposure and other environmental stresses, Or extract of the present invention is useful, and has found efficacy in anti-aging including anti-inflammation, wound healing, skin barrier enhancement, skin convergence, wrinkle improvement, and elasticity improvement.

Accordingly, an object of the present invention is to provide a method for producing the present invention provides a composition for external application for skin for improving skin comprising plant cells, adventitious roots or extract thereof as an active ingredient.

In the present invention, the skin improving agent includes skin aging, wrinkle improving, skin barrier function protecting or strengthening, and skin elasticity enhancement.

It is another object of the present invention, Ulleungdo native plant haeguk (Aster sphatholifolius Maxim.) as an active ingredient, or an extract thereof.

Ulleungdo native plant decline ( Aster spha ntulifolius Maxim.) or an extract thereof as an active ingredient.

It is still another object of the present invention to provide a method for preparing a skin external application composition for skin improvement, anti-inflammation or wound healing, which comprises the plant cell, the adventitious root culture of the Ulleungdo native plant or the extract thereof as an active ingredient.

In order to achieve the above object, the present invention provides a composition for external application for skin for improving skin comprising plant cells, adventitious roots of Aster sphathulifolius Maxim. Or extract thereof as an active ingredient.

The present invention also relates to a method for producing a microorganism, sphatholifolius Maxim.) plant cell, adventitious root culture or extract thereof as an active ingredient.

The present invention also relates to a method for producing a microorganism, The present invention provides a composition for external application for skin wrinkles containing plant cells, adventitious roots or extract thereof as an active ingredient.

The present invention also relates to a method for producing a microorganism, The present invention provides a skin barrier or a skin external application composition for enhancing a skin barrier comprising plant cells, adventitious roots or extract thereof as an active ingredient.

The present invention also relates to a method for producing a microorganism, The present invention provides a skin external application composition for promoting skin elasticity comprising plant cells, adventitious roots cultures or extracts thereof as an active ingredient.

The present invention also relates to a method for producing a microorganism, sphatholifolius Maxim.) as an active ingredient, or an extract thereof.

The present invention also relates to a method for producing a microorganism, The present invention provides a composition for external application for wound healing, which contains plant cells, adventitious roots cultures or extracts thereof as an active ingredient.

The present invention also relates to a method of producing an Aster The present invention provides a method for preparing a skin external application composition for skin improvement, anti-inflammation or wound healing, which comprises a culture of a cell of a thyroid cell of a plant of sphathulifolius Maxim. or an extract thereof,

(a) separating the leaves, stems, petals, or embryonic placenta in the ovary plant and culturing them into plant cells or adventitious roots; And (b) preparing an external composition for skin containing the cultured Korean plant cell or adventitious root culture or extract thereof.

The present invention relates to a composition for external application of a plant cell or adventitious roots or an extract thereof, which is free from toxicity to skin cells and has excellent skin collagen synthesis and skin barrier protection and skin collagen synthesis ability, , Anti-inflammation, and wound healing efficacy.

FIG. 1 is a diagram showing the process of inducing plant cells from offshoot plants.
2 is a graph showing the results of HPLC analysis of plant cell culture extracts derived from offshoot plants.
FIG. 3 shows experimental results for confirming the skin barrier protection function of plant cell culture extracts derived from offshoot plants.
FIG. 4 shows the results of an experiment in which COX-2 and iNOS expression were increased.
Fig. 5 shows experimental results for confirming the wound healing function of plant cell culture extracts derived from offshoot plants.
6 is a graph showing the healing area in order to confirm the wound healing function of plant cell culture extracts derived from offshoot plants.

The present invention relates to a composition for external application for skin containing plant cells or adventitious roots of Aster sphathulifolius Maxim. Or an extract thereof as an active ingredient, and a process for producing the same.

In one aspect, the present invention relates to a skin external composition for improving skin comprising plant cells or adventitious roots of Aster sphathulifolius Maxim. Or an extract thereof as an active ingredient.

In the present invention, the term "for skin improvement" is intended to include improvements in the condition of the skin such as skin convergence, antiaging, skin wrinkle improvement, skin elasticity enhancement, skin barrier function protection or enhancement.

In the present invention, the term "comprising as an active ingredient" means a composition for external application for skin, which is capable of exhibiting various skin improving effects, antiinflammatory and wound healing properties as described above, Quot; means an amount of effective amount that is capable of exhibiting synthetic enhancement, the ability to express elastase in association with the enhancement of elasticity, the increase in pilar green expression associated with protection of skin barrier function, and the like.

In the present invention, the cultured offspring plant cell culture is obtained by culturing a part of the offspring plant, for example, whole or part of the stem, petal, roots, leaves, and placenta in a cut-off induction medium.

Such culture induction is related to plant tissue culture, and plant tissue culture is a technique for cultivating asphyxiated small plant tissue in vitro to grow plant cells, organs and plants according to the purpose of culture. Plant tissue culture can be attributed to the basic characteristics of plants. Plant tissue culture technology is based on the use of differentiating totipotency to regenerate plants from plant somatic cells, a unique feature of plants. Namely, plants can be regenerated from these cells and tissues by culturing single cells (including protoplasts), leaves and root sections of plants in a medium supplemented with specific nutrients and growth regulators. The ability of these plants to have differentiating totipotency is a strategy for survival from harsh environments and herbivorous damage for the fixation and long-term survival of plants. Therefore, plants can be said to have the ability to regenerate lost organs again. The plant tissue cells isolated from the parent plant have the potential to be reproduced as a complete plant when cultured in a suitable culture environment, that is, plants having typical performance are of academic importance such as embryology, but they are also utilized for practical use of cosmetic raw materials .

"Plant cell culture" refers to a specific tissue or cell mass produced when a tissue cut out from a plant is cultured in a medium containing an auxin, a wound is injured to a certain kind of plant, or an auxin is treated with the excipient. Usually callus is an amorphous tissue or cell mass that has lost its ability to cause normal organogenesis or tissue differentiation, and is mostly flowcytosed. In a broad sense, it also includes plant tumor tissue caused by infection such as agrobacterium.

Therefore, it is possible to induce any part derived from any part such as stem, leaf or the like according to the present invention. However, in one embodiment, the cotyledon is cut out from the stem of the decollete and a part of the leaf or germinated seedlings of the seedlings Derived plant cell, and plant cells may be formed by cutting off some of the tissues of the decidua and placing the content of auxin and cytokinin in a regulated medium. In the case of adventitious root cultivation, the same process as that of plant cell cultivation is carried out, but the combination of plant hormones is different so that the adventitious roots can be induced. For example, if the plant hormone contained in the induction medium is a plant cell culture, a combination of p-chlorophenoxyacetic acid and kinetin, or in the case of adventitious roots, p-chlorophenoxyacetic acid and indolebutyric acid are contained and induction-cultured.

The placenta of the plant is an important part that regulates seed breeding, such as an animal-derived placenta. It is a part of the ovary in the pistil (ovary, ovary). It is usually on the edge of the carpel, which forms the ovary. It also occurs in the base of the ovary, or in the shape of a column standing upright in the ovary from the base. The laver of the plant is structurally derived from the ovary, a part of the ovule connected to the ovary, from the floral meristem. The undergrowth develops from the plant head. To cultivate the embryonic cells of the embryo, carefully remove the ovary wall from the bud using tweezers and separate the embryonic cells.

In the present invention, the plant cell or the adventitious root culture or the extract thereof may be contained in an amount of 0.1 to 10% by weight based on the total weight of the composition.

These ratios are merely preferable ranges. For example, in the following examples, 1 wt% or 2 wt% of the test results are included. However, it is confirmed that 5% and 10% Survival rate was also not affected by 10%. It is apparent to those skilled in the art that, in addition to 20% and 30% or more, excellent skin improving effect, reinforcement of skin barrier, improvement of skin, convergence of skin, improvement of wrinkles, anti-inflammation and wound healing function.

In the present invention, the skin improving agent includes skin irritation, wrinkle improving, skin barrier strengthening or improvement.

In the present invention, the haeguk (Aster sphathulifolius Maxim.), the cultured material may be used by filtration, the culture may be used after being crushed, or the culture may be dried to be powdered and then dissolved in purified water. have.

In the present invention, the "extract" may be obtained by pulverizing the plant cell or adventitious root culture extract of Aster spha tulifolius Maxim. Or by extracting it by various methods known in the art such as cold water extraction method, hot water extraction method, ethanol and jojoba oil extraction method Lt; / RTI >

The extraction method in the present invention is not particularly limited, and examples thereof include cold extraction, ultrasonic extraction, reflux extraction, hot water extraction, and jojoba oil extraction. Here, in the case of hot water extraction, hot water extract is obtained by heating at 80 to 100 ° C for 8 to 48 hours in a hot water distiller.

In the case of cold water extraction, for example, cold water (15 to 25 ° C) is mixed with the culture itself or the dried powder thereof and extracted for 3 days to obtain a cold water extract.

Alternatively, it can be produced by a method of extracting using water, an organic solvent, or a mixed solvent thereof. The extracted liquid can be used directly or by concentrating and / or drying. In the case of extraction using an organic solvent, an organic solvent such as methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, (DMSO), 1,3-butylene glycol, propylene glycol, or a mixed solvent thereof. The active ingredient of the herbal medicine may be extracted at room temperature or warmed under the condition that the active ingredient is not destroyed or minimized. Depending on the organic solvent to be extracted, the degree of extraction and the degree of loss of the active ingredient of the medicament may differ. Therefore, an appropriate organic solvent should be selected and used. Particularly, in the present invention, an ethanol extract, preferably a 20-50% ethanol extract, specifically 50% ethanol extract, is preferable. As described in the examples, the extract is mixed with 50% ≪ / RTI >

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

The invention from another point of view, haeguk (Aster The present invention provides a method for preparing a skin external application composition for skin improvement, anti-inflammation or wound healing, which comprises a culture of a cell of a thyroid cell of a plant of sphathulifolius Maxim. or an extract thereof,

(a) separating leaves, stems, petals, or placental tissues in ovary of a native plant and culturing them into plant cells or adventitious roots; And

(b) a step of preparing an external skin application composition containing the cultured plant cell culture or adventitious root culture or extract thereof.

The step (a) is a step of obtaining a plant tissue culture, that is, a plant cell culture, or an adventitious culture, with some tissue separated from the native plant.

In the present invention, leaves, stems, petals, roots, or oocytes in the ovary can be separated and used.

In the present invention, a suitable medium may be selected for deriving the plant cell culture or the adventitious root culture from the separated tissue. In the step (a), a suitable medium may be selected to cultivate the transformed plant cells or adventitious roots. If there is a medium commonly used in plant tissue culture in the technical field, it can be used without limitation. For example, the composition of the MS medium (1 L standard) is 1650 mg NH ₄ NO ₃ , 1900 mg KNO ₃ , 440 mg CaCl ₂ .2H ₂ O, MgSO 4, _{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, and Sucrose 30000 mg.

Although there is almost no difference in the basic MS medium composition during the induction of plant cell culture from plant leaves, plant stem, plant taiga, and plant petal origin, the kinds of plant growth hormone that are most important in inducing plant cells and adventitious roots are different Do.

In the case of the plant cell culture according to the present invention, in the case of plant cell culture derived from a leaf or a stem, a plant cell culture can be obtained by culturing in the presence of p-chlorophenoxyacetic acid and kinetin have.

The combination ratio of each hormone may be 1: 0.02 to 0.1, or 1: 0.05 to 0.08 for p-chlorophenoxyacetic acid and kinetin.

In the plant cell culture according to the present invention, plant cell cultures can be obtained by culturing plant cell cultures derived from plant placenta tissues containing p-chlorophenoxyacetic acid and zeatin. have.

The combination ratio of each of the hormones may be characterized by being 1: 0.1-0.5, or 1: 0.2-0.4 in terms of p-chlorophenoxyacetic acid and zeatin.

On the other hand, in the case of adventitious root culture, the adriamycin culture can be obtained by culturing it in the medium containing p-chlorophenoxyacetic acid and indolebutyric acid (IBA).

The combination ratio of each hormone may be 1: 1 to 4, or 1: 2 to 3 for p-chlorophenoxyacetic acid and indole butyric acid (IBA).

In the present invention, it is preferable that in the step (b), the composition containing the plant cell or the adventitious root culture obtained as a result of the cultivation in the step (a) is prepared from a suitable external preparation for skin composition, And can be prepared in the form of an external preparation for skin in a suitable form.

For example, in step (b), the plant cell or adventitious root culture obtained in step (a) is dried, powdered and mixed with purified water to prepare a composition,

the plant cell or adventitious root culture obtained in step (a) is dried, powdered and mixed with purified water, and then extracted with cold water, hot water, jojoba oil, vegetable oil, inorganic solvent or ethanol And then extracting the solution with an organic solvent such as alcohol to prepare a composition.

As another example, the culture obtained in the step (a) may be dried by hot air and powdered to evaporate water.

In the present invention, the principle of skin astringent action is a phenomenon in which skin is contracted by binding with polymer flavonoids of skin protein to form crosslinks. Convergence means basically wrinkled or funky. Convergent agent has a function to shrink skin and mucosal blood vessels, and has an effect of inhibiting secretion of mucus by interrupting cell gap and lymphatic gap. In addition, since astringent agent has a property of binding to protein, it can generally judge degree of convergence effect according to degree of binding of hemoglobin protein to extract. Such convergent action may be said to have an effect of forming an insoluble film on the surface of the skin and mucous membranes by external application or contents and consequently protecting the locality or densifying the tissue to reduce the permeability of the cell membrane.

In the skin barrier protection function according to the present invention, the skin barrier is composed of a stratum corneum, which is the outermost layer of the epidermis, and a corneocyte, which is mainly non-nucleated. The multilamellar lipid layer, composed of intercellular lipids such as ceramides, cholesterol, and fatty acids synthesized by keratinocytes of the skin barrier maintained through normal division and differentiation of epidermal cells, It plays a role. Meanwhile, omega hydroxyceramide in the intercellular lipids is chemically covalently bound to involucrin, which is a protein of the corneocyte outer layer, to form a corneocyte lipid envelope (CLE) Type liposomes to physically stabilize the intercellular lipids, thereby enhancing and enhancing the barrier function.

The composition of the present invention is delivered to the stratum corneum through skin application to promote the differentiation of keratinocytes, thereby improving the thickness of the skin layer. In addition, the composition has excellent effect of restoring skin barrier damage, And can be usefully used for the treatment and prevention of skin diseases caused. Skin diseases caused by skin barrier damage include, but are not limited to, atopic dermatitis, xeroderma, psoriasis, ichthyosis, acne and the like.

In addition, the mechanism of skin barrier protection was confirmed by the increase of the protein amount of occludin and claudin-1, which are close-coupled conformation proteins. Filaggrin is one of several structural proteins that keratinocytes express at the differentiation stage. It is involved in differentiation from the basal layer to the stratum corneum of the epidermis and is a natural moisture factor essential for the maintenance of the moisture of the skin tissue. NMF), and is used as a main index of moisture retention and skin membrane function of the skin. However, since the gene expression of fila green is remarkably increased, the Korean native plant cell, the adventitious root culture or its extract according to the present invention, Barrier protection, enhancement, and improvement.

In addition, the transformed plant cell, adventitious root culture or extract thereof according to the present invention has elastin expression ability. Elastin is a structural substance that exists in the extracellular matrix of connective tissue and is a substance that is present in the skin and imparts elasticity. Elastin is synthesized in cells with a polypeptide called Tropo elastin and then cross-linked between the tropoelastins outside the cell. Thus, elastin has a secondary or three-dimensional elasticity due to cross-linking of tropoelastin. Therefore, the plant cell of the present invention, the cultured adventitious roots or the extract thereof can be utilized for enhancing and improving skin elasticity.

In addition, the plant cells, adventitious roots or extracts thereof according to the present invention promote the proliferation and migration of keratinocytes, and have wound healing and anti-aging effects.

In addition, the offspring plant cells, adventitious root cultures or extracts thereof according to the present invention have anti-inflammatory effects through inhibition of COX-2 and iNOS activity.

In addition, the composition for external application for skin according to the present invention can increase the amount of pro-collagen and collagen biosynthesis, and can be applied to prevent skin aging and improve / prevent skin wrinkles.

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

The cosmetic composition of the present invention may contain an acceptable carrier in cosmetic preparations. Herein, "an acceptable carrier for a cosmetic preparation" is a known or used compound or composition that can be contained in a cosmetic preparation, or a compound or composition to be developed in the future, which is toxic, instable or irritating It says 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 composition for external application for skin according to the present invention may contain glycerin, butylene glycol, propylene glycol, polyoxyethylene hydrogenated castor oil, ethanol, triethanolamine, etc., And preservatives, antioxidants, coloring agents, purified water, and the like may be included as needed.

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 producing a dermal plant cell, an adventitious root culture or an external composition for skin containing the extract according to the present invention is not limited to the above-described manufacturing method, and any person skilled in the art will recognize that, May also be used to prepare a composition for external application for skin containing a Korean native plant cell, an adventitious root culture, or an extract thereof according to the present invention.

In particular, the composition for external application for skin may be prepared in the form of a general emulsified formulation and a solubilized formulation, by a known manufacturing method, in addition to the manufacturing method specifically disclosed in the present invention.

When the composition is prepared with a composition for external application for skin, examples of the cosmetic composition of the emulsified form include nutritional lotion, cream, essence and the like. It can also be prepared in the form of adjuvants which can be applied topically or systemically, which are conventionally used in the field of dermatology by containing a dermatologically acceptable medium or base.

In addition, suitable cosmetic formulations include, for example, solutions, gels, solid or kneaded anhydrous products, emulsions obtained by dispersing an oil phase in water, suspensions, microemulsions, microcapsules, microgranules or ionic forms (liposomes) May be provided in the form of a follicular dispersing agent of the type, cream, skin, lotion, powder, ointment, spray or conical stick. It can also be prepared in the form of a foam or an aerosol composition further containing a compressed propellant.

In addition, the composition for external application for skin of the present invention may further comprise at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softening agents, antioxidants, suspending agents, stabilizers, foaming agents, As used herein means any emulsion or emulsion which is commonly used in emulsifying or emulsifying agents, fillers, sequestering and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics Adjuvants commonly used in the cosmetics or dermatological fields, such as other ingredients. And, the above ingredients can be introduced in amounts commonly used in the field of dermatology.

The composition for external application for skin according to the present invention is useful for preventing skin aging due to the ability of promoting collagen synthesis, improving skin elasticity by enhancing expression of elastase, protecting skin barrier function, enhancing skin barrier function, improving antiinflammation, And functional cosmetics capable of protecting or enhancing skin barrier function.

In the present invention, in the case of a pharmaceutical composition, it can function as a pharmaceutical composition for skin condition improvement as shown in the following examples.

Such a pharmaceutical composition may contain, in addition to the active ingredient, a corn plant cell, an adventitious root culture or an extract thereof, a "pharmaceutically acceptable carrier ", which carrier may be a diluent, a lubricant, a binder, a disintegrant, And a preservative. 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 into conventional pharmaceutical formulations known in the art. The pharmaceutical composition may be formulated and administered in the form of an oral administration preparation, an injection preparation, a suppository, a transdermal preparation, and a dosage form. For example, the formulations may be formulated for oral administration, such as solutions, suspensions, powders, granules, tablets, capsules, pills, or expanses.

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

Particularly, in the following examples, the efficacy with respect to cultured plant cells, adventitious roots or extracts thereof was confirmed, but it will be apparent to those skilled in the art that such effects are also obtained with respect to the culture itself, not the extract.

1-1. Cell induction from offshoot plants according to the present invention

1.1 Induction of Plant Cells in Offspring Plants

The offshore plants were sterilized for 5 minutes with 70% ethanol and 0.5% sodium hypochlorite, and washed several times with sterile water. Sucrose 30.0 g / L, Agar 5 to 10.0 g / L, p-Chlorophenoxyacetic acid (Mucin) were added to the MS basal medium (Murashige and Skoog 1962, Duchefa, Cat No. M0221) (4-CPA) 2 mg / L, Kinetin 0.1 mg, pH 5.5 to 6.0, preferably pH 5.7 to 5.8. Approximately four weeks later, the offspring cotyled on the medium induced cotransplant plant cells. Induced native plant cells were proliferated by subculture at 2 - week intervals.

1.2 Induction of Tacrolimus Cells in Offspring Plants

The offshore plants were sterilized for 5 minutes with 70% ethanol and 0.5% sodium hypochlorite, and washed several times with sterile water. Sucrose 30.0 g / L, Agar 5 to 10.0 g / L, p (p <0.05) were added to the MS basal medium (Murashige and Skoog 1962, Duchefa, Cat No. M0221) by carefully removing the ovary wall with the use of tweezers. The cells were subjected to MS solid medium at pH 5.5 to 6.0, preferably pH 5.7 to 5.8, containing 2 mg / L of chlorophenoxyacetic acid (4-CPA) and 0.5 mg of Zeatin. Tumor cells were induced by culturing for 2 ~ 3 weeks. The induced cells were proliferated by subculture at 2 - week intervals.

1.3 Derivation of adventitious roots of domesticated plants

The offshore plants were sterilized for 5 minutes with 70% ethanol and 0.5% sodium hypochlorite, and washed several times with sterile water. Sucrose 30.0 g / L, Agar 5 to 10.0 g / L in MS basal medium (Murashige and Skoog 1962, Duchefa, Cat No. M0221) was carefully cut with a tweezers and a sharp knife at once, was applied to MS solid medium at pH 5.5 to 6.0, preferably pH 5.7 to 5.8, containing 2 mg / L of p-Chlorophenoxyacetic acid (4-CPA) and 5 mg of Indole-3-butyric acid (IBA). Adrenal ganglia were induced by culturing for 2 to 3 weeks. Induced dorsal root ganglia were propagated by subculture at 2 - week intervals.

1-2. Plant cells, embryonic stem cells, adventitious root culture and mass production derived from offshoot plants according to the present invention

Plant cells, embryonic stem cells and adventitious roots of aseptically induced offspring plants were cultured on MS medium containing 3% sucrose and 2 mg / L of p-chlorophenoxyacetic acid (4-CPA) and 0.1 mg of Kinetin for a period of 2-3 weeks , And then cultured in a liquid culture medium of the same composition except agar using a balloon type bioreactor (Samsung Science) at a temperature of 25 ° C. and an air supply amount of 0.1 vvm, Respectively.

1-3. Production of Plant Cells, Tacrine Cells, and Root Extracts Derived from Seaweed Plants According to the Present Invention

The proliferated cultured plant cells, thyroid cells and adventitious roots were harvested, and then the water was removed with a clean tissue and dried in a dryer at 60 ° C for 2 days. 100 g of the dried culture was placed in a container, and 10 L of purified water (or 10 L of 70% ethanol) was added thereto, followed by heating at 80 to 100 캜 for 8 to 48 hours in a hot water distiller to obtain a hot water extract. After the extraction, the extract was filtered with a mesh to remove solid components, and thus the extracts of Daejuk plant cell cultures, Taejung cell cultures and Rhizome roots extracts were prepared.

1-4. Analysis of HPLC components of culture extracts

Component analysis was carried out by HPLC on cultured plant cells, embryonic stem cells, and adventitious root culture extracts. The analysis conditions were as follows.

division Analysis condition HPLC device type Waters 1525 Binary HPLC Pump Column type Gemini 5 C18 110 A (Phenomenex) Column Specification 250 * 4.60 mm, 5 micron Solvent conditions - Solvent A: Water (containing 0.1% TFA (Trifluoro acetic acid))
- Solvent B: Acetonitrile (containing 0.1% TFA (Trifluoro acetic acid)) wavelength 234 nm (UV lamp) Flow rate 1 ml / min

As a result of the analysis of the extract components of the plant cell culture, HPLC analysis peak results as shown in FIG. 2 were obtained. Also, it was confirmed that the hot water extract, ethanol extract, hydrothermal extract and ethanol extract of turmeric cell culture had similar peak values.

Experimental Example 1: Skin Convergence Effect Test of Dissolved Plant Cells, Tacrine Cells and Adventitious Root Culture Extracts According to the Present Invention

The principle of convergence is the phenomenon that the skin protein is contracted by binding with polymer flavonoids to form crosslinks. Convergence means basically wrinkling or retention, and since astringent agent has a property of binding to protein, it can generally judge the convergence effect depending on the degree of binding of hemoglobin protein to the extract. Such convergent action may be said to have the effect of forming an insoluble film on the surface of the skin and mucous membrane by external action and densifying the tissue to reduce the permeability of the cell membrane. For measurement of convergence effect, blood protein (hemoglobin, Sigma, Cat. # 08449) similar to skin protein was used. Each sample solution and hemoglobin solution (500ppm) were mixed in a ratio of 1: 1, shaken, and centrifuged at 1,500 rpm for 3 minutes, and the absorbance was measured at 576 nm. The measurement of the convergence effect was expressed by the absorbance reduction rate of the sample and the control group.

[Equation 1]

Concentration effect (%) = (1-absorbance of sample-added group / absorbance of no-addition group) x 100

Treatment concentration Measuring Convergence Effect
(Astringent activity test)
(% of control) Control group - 0 Tannic
acid - 37.9 Heat number
extraction Plant cell
Culture extract 0.5% 1.5 One % 13.2 5% 30.1 10% 34.0 Teat cell
Culture extract 0.5% 0.3 One % 14.2 5% 22.1 10% 28.0 Adrenaline
Culture extract 0.5% 1.1 One % 9.3 5% 20.3 10% 30.1 ethanol
extraction Plant cell
Culture extract 0.5% 2.5 One % 15.2 5% 38.1 10% 40.0 Teat cell
Culture extract 0.5% 1.3 One % 13.2 5% 26.1 10% 31.0 Adrenaline
Culture extract 0.5% 1.1 One % 10.3 5% 22.3 10% 34.1

As a result of measuring the convergence effect of plant cell, embryonic cell and adventitious root culture extracts, it was confirmed that the concentration - dependent convergence effect was excellent in leaf - derived plant cell, embryonic cell and adventitious root culture extracts compared to the control group.

Experimental Example  2: Procollagen  Synthetic enhancement test

Human fibroblasts (Human Skin Fibroblast) for 37 ℃, 5% in a CO ₂ incubator of 10% in the cell culture medium to maintain a constant humidity FBS, Penicillin (50U / ml) , Streptomycin DMEM (Dulbecco's containing (50 / ml) (Gibco, USA). Cells were seeded at a density of 1 × 10 ⁵ cells / ml in a 24-well plate and cultured for 24 hours. The control (DMEM medium) and the medium containing the test substance in the concentration were added and cultured in a 5% CO ₂ incubator at 37 ° C for 48 hours. After 48 hours, the amount of newly synthesized collagen was measured by measuring 20 μl of each supernatant of the culture medium and measuring it using Procollagen Type I C-Peptide EIA Kit (PICP, Takara, Cat No. MK101). The amount of PICP was converted into ng / 1 x 10 &lt; ⁵ &gt; cells, calculated according to the following formula, and the results are shown in Table 3.

&Quot; (2) &quot;

Treatment concentration Procollagen biosynthetic amount
PIP (ng / ml) Negative control group - 70 Positive control group
TGF-1
(10 ng / ml) - 280 Heat number
extraction Plant cell
Culture extract 0.5% 75 One % 85 5% 108 10% 118 Teat cell
Culture extract 0.5% 75 One % 210 5% 234 10% 240 Adrenaline
Culture extract 0.5% 87 One % 197 5% 220 10% 195 ethanol
extraction Plant cell
Culture extract 0.5% 154 One % 187 5% 205 10% 221 Teat cell
Culture extract 0.5% 94 One % 120 5% 234 10% 200 Adrenaline
Culture extract 0.5% 70 One % 84 5% 120 10% 151

As a result of measuring the amount of procollagen production related to the collagen synthesis of the plant cell, teat cell, and adventitious root culture derived from the seawater, the extractive and ethanol extracts from the seaweed - derived plant cell, And it was confirmed that the effect of procollagen biosynthesis was excellent.

Experimental Example  3: Skin barrier protection mechanism confirmation test

We examined whether skin barrier enhancement and enhancement functions by checking the mechanism of protection of skin barrier by plant cell, turmeric cell and adventitious root culture extracts of the country.

Human epidermal keratinocytes were incubated with Dulbecco's Modified Eagle's Medium (DMEM), 10% Fatal bovine serum (FBS) and 1% Antibiotic-Antimycotic (GIBCO, Cat. # 15240-062) ² culture dishes at 37 ° C and 5% CO ₂ .

When the human epidermal keratinocytes were confluent at a concentration of 80% or more, they were divided into 12-wells at 3 × 10 ⁵ cells / well and cultured for 48 hours. UVB 40 mJ / cm ² was irradiated using an ultraviolet light irradiator, And cultured in a medium containing 1% each for 3 days. After the incubation, the cells were eluted with cell lysis buffer (0.1% SDS, 1% NP40, 150 mM NaCl, 0.5% sodium deoxycholate, 50 mM Tris-Cl (pH 7.5)) and quantitated by bicinchoninic acid Were measured. Twenty grams of the same amount of protein was added, separated by SDS-polyacrylamide gel electrophoresis, and transferred to a nitrocellulose membrane. Membrane was incubated for 1 h with TBST (10 mM Tris HCl, pH 8.0, 150 mM NaCl, 0.1% Tween 20) containing 5% skim milk and then incubated with occludin (Invitrogen, USA), claudin-1 (abcam, USA) And β-actin (Santa Cruz, USA) primary antibodies. The secondary antibody was reacted with goat anti-rabbit IgG-HRP (Santa Cruz, USA) and donkey anti-goat IgG-HRP (Santa Cruz, USA) for 1 hour. After washing with TBST buffer, the cells were developed with ECL solution kit (Amersham, UK).

To investigate the quantitative changes of adhesion - conformational proteins by plant - derived cells, teatocytes and adventitious roots extracts, we examined the protein changes using keratinocytes. As shown in FIG. 3, keratinocytes irradiated with ultraviolet light showed reduced amounts of occludin and claudin-1 proteins, which are closely related to each other. However, after treatment with ultraviolet radiation, plant cells, embryonic stem cells and adventitious root culture extracts Decreased occludin protein production was increased.

Experimental Example  4: Skin barrier function test

Human epidermal keratinocytes were incubated with Dulbecco's Modified Eagle's Medium (DMEM), 10% Fatal bovine serum (FBS) and 1% Antibiotic-Antimycotic (GIBCO, Cat. # 15240-062) ² culture dishes at 37 ° C and 5% CO ₂ .

When the human epidermal keratinocytes were confluent at 80% or more, the cells were cultured in a 12-well plate at 3 × 10 ⁵ cells / well for 24 hours and then cultured in a culture medium containing 1% of the test substance for 24 hours. When more than 80% confluence of human epidermal keratinocytes is achieved, the cells are seeded at a density of 1 × 10 ⁴ cells / well in a 96-well plate. Then, the plant cells, embryonic cells and adventitious culture extracts of the embryonic cells are treated. After 24 hours of incubation, the expression of filaggrin gene was confirmed by real-time PCR.

RNA isolation and cDNA synthesis are performed using FastLane Cell one-step buffer set (QIAGEN, Cat. # 216213). The cells were washed with FCW (cell wash buffer), and incubated at room temperature for 5 min with 50 μl cell processing mixture (Buffer FCPL supplemented with Buffer FCPL and gDAN Wipeout buffer) And reacted at 75 ° C for 5 minutes. In order to analyze the expression of the filaggrin gene, the cDNA synthesized above is used as a template and subjected to a Rotor Gene Q Real-time PCR Machine (Qiagen) according to the manual of 2X QuantiTect SYBR Green RT-PCR Master Mix. The filaggrin primer used in the experiment was Qiagen Cat. # QT01192646) and the expression rate was Normalization with Housekeeping genes (GAPDH, Cat. # QT01192646). Real-time PCR conditions are as follows.

Treatment concentration sample Filaggrin gene expression rate
(Relative Expression of Filaggrin)  Negative control group 1.00 One % Hot water extraction Plant cell 2.52 Teat cell 1.98 Adrenaline 1.41 Ethanol extraction Plant cell 2.38 Teat cell 3.38 Adrenaline 1.91

The increased expression of filaggrin promotes the normal differentiation of keratinocytes by plant-derived cells, embryonic stem cells and adventitious roots extracts, and promotes the expression of filaggrin in the skin, which is converted into a natural moisturizing factor in the skin. Can be improved.

Experimental Example  5: Analysis of elastin gene expression

Human epidermal fibroblasts were incubated with 100 mM / 60.1 cm ² of Dulbecco's Modified Eagle's Medium (DMEM), 10% Fatal bovine serum (FBS) and 1% Antibiotic-Antimycotic (GIBCO, Cat. # 15240-062) culture dishes at 37 ° C and 5% CO ₂ .

When human dendritic cells were confluent at a concentration of 80% or more, cells were seeded at 3 × 10 ⁵ cells / well in a 12-well plate and cultured for 24 hours. Then, plant cells, embryonic stem cells and adventitious root culture extracts contained 1% Lt; / RTI &gt; for 24 hours. When human dendritic cells are confluent at a concentration of 80% or more, they are seeded at 1 × 10 ⁴ cells / well in a 96-well plate and then treated with the plant cell, embryonic cell and adventitious culture extract of the embryo. After that, the cells were further cultured for 24 hours and Elastin gene expression was confirmed by Real-time PCR.

 RNA isolation and cDNA synthesis are performed using FastLane Cell one-step buffer set (QIAGEN, Cat. # 216213). The cells were washed with FCW (cell wash buffer), and incubated at room temperature for 5 min with 50 μl cell processing mixture (Buffer FCPL supplemented with Buffer FCPL and gDAN Wipeout buffer) And reacted at 75 ° C for 5 minutes. In order to analyze the expression of the filaggrin gene, the cDNA synthesized above is used as a template and subjected to a Rotor Gene Q Real-time PCR Machine (Qiagen) according to the manual of 2X QuantiTect SYBR Green RT-PCR Master Mix. The Elastin primer used in this study was Qiagen Cat. # QT00034594) and the expression rate was Normalization with Housekeeping genes (GAPDH, Cat. # QT01192646). Real-time PCR conditions are as follows.

Treatment concentration part Elastin gene expression rate
(Relative Expression of Elastin)  Negative control group - 1.00 Positive control group
TGF-1
(10 ng / ml) - 1.52 One % Hot water extraction Plant cell 1.29 Teat cell 1.42 Adrenaline 1.39 Ethanol extraction Plant cell 1.19 Teat cell 1.38 Adrenaline 1.31

The expression of Elastin gene related to the skin elasticity of the plant cell, the embryonic cell and the adventitious root culture derived from the seawater showed that the expression rate of Elastin gene was increased by the plant cell, Respectively.

Experimental Example  6: COX -2 and iNOS  Anti-inflammatory efficacy test through active inhibition

Human keratinocyte cell line HaCaT (human keratinocyte) to Dulbecco's Modified Eagle's Medium (DMEM ), 10% Fatal bovine serum (FBS), 1% Antibiotic-Antimycotic (GIBCO, Cat. # 15240-062) and 100 mm / 60.1 cm ² with culture dishes at 37 ° C and 5% CO 2. When HaCaT was confluent at 90% confluence, 1 × 10 ⁵ cells / dish was dispensed into 100 mm / 60.1 cm ² culture dishes, and cultured for 24 hours. After incubation for 4 hrs, the cells were lysed with 0.1% SDS, 1% NP40, 150 mM NaCl, 0.5% sodium deoxycholate, 50 mM Tris-Cl (pH 7.5) and protease inhibitors (Roche, Mannheim, Germany) And protein was quantitated by BCA assay. After transferring the separated proteins to the NC membrane, the membrane was blocked with blocking solution (5% skim milk in TBST (Tris-buffered saline with 0.1% Tween 20 ), Blocked for 30 minutes or more, washed with TBST, and incubated overnight at 4 ° C in a primary antibody (COX-2, Abcam, Cat. # Ab-6665) solution. Then, the cells were washed 3 times for 10 minutes with TBST and reacted with primary antibody conjugated secondary antibody (COX-2: rabbit, iNOS: mouse) for 2 hours. The blot was analyzed by ECL solution kit (Amersham, UK).

The effect of COX-2 and iNOS on the expression of COX-2 and iNOS was investigated by UV irradiation. As a positive control, 5 uM hydrocortisone was used. As a result, as shown in FIG. 4, it was confirmed that the expression of COX-2 and iNOS was decreased in all of the plant cells, tubercle cells and adventitious root culture extracts by hot water extraction and ethanol extraction.

Experimental Example  7: Wound healing assay Using Wound healing , Anti-aging efficacy test

Human keratinocyte cell line HaCaT (human keratinocyte) was incubated in 100 mm / 60.1 cm ² culture dishes at 37 ° C and 5 ° C with Dulbecco's Modified Eagle's Medium (DMEM), 10% Fatal bovine serum (FBS) and 1% Antibiotic-Antimycotic % CO2. When HaCaT was confluent at 90% confluence, cells were plated at 1.5 × 10 ⁵ cells / 12-well plate in 100 mm / 60.1 cm ² dish and cultured for 24 hours. After replacing with medium containing no FBS, And treated with 1% of test substance, plant cells derived from offshoot plants, embryonic stem cells and adventitious root culture extract, respectively. The cell image was photographed at 0 hour after material treatment and cultured at 37 ° C and 5% CO 2. After 18-20 h incubation, the medium was removed and incubated for 15 min at room temperature in a fixing solution (4% paraformaldehyde) and washed 3 times with PBS. Fixed cells were stored at 4 ° C for one month. The extent of the wound healing was measured by taking a picture under a microscope and using Image J program. The positive control group was 300 ng / ml EGF.

 Wound healing assay was used to evaluate the antioxidant effect by observing whether or not the cell proliferation and migration were promoted by the test substance. As a positive control, 300 ng / ml EGF was used.

As shown in FIG. 5 and FIG. 6, the extracts of plant cells, turmeric cells and adventitious roots derived from seaweed were respectively treated at 1% concentration. As a result, in both hot water extraction and ethanol extraction, As a result, it was confirmed that the healing area was increased by adhering to the skin cells and the floor.

Preparation of composition for external application for skin

Cosmetics of the emulsified form such as nutrient lotion, cream, essence, etc. and cosmetics of the solubilized form such as the softened longevity were prepared by the cosmetic product containing the exported plant,

Production Example 2-1: Lotion

According to the following formulation, it was prepared according to the conventional lotion preparation method.

Raw material name Weight% (w / w) glycerin 5.0 Dipropylene glycol 3.0 Hyaluronic acid 0.5 Polyoxyethylene hardened castor oil 0.1 Polyethylene oleyl ethyl 0.1 ethanol 5.0 antiseptic 0.15 Beauty Suitable amount flash Suitable amount Origin plant, Taejung, Root culture extract 2.0 Purified water to 100

Production Example 2-2: Essence

According to the following prescription, it was prepared according to a conventional method for producing essence.

Raw material name Weight% (w / w) Cetostearyl alcohol 1.0 Self emulsifying monostearic acid 1.0 Wax 0.5 Squalane 5.0 Isocetyl octanoate 3.0 Dimethylsiloxane 0.3 Sorbitan monostearate 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 Origin plant, Taejung, Root culture extract 7.0 Purified water to 100

Production Example 2-3: Lotion

According to the following formulation, it was prepared according to a conventional lotion preparation method.

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 Isocetyl octanoate 2.0 Dimethylsiloxane 0.3 Sorbitan monostearate 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 Origin plant, Taejung, Root culture extract 5.0 Purified water to 100

Production Example 2-4: Cream

According to the following formulation, it was prepared according to a conventional cream production method.

Raw material name Weight% (w / w) Cetostearyl alcohol 3.0 Self emulsifying monostearic acid 1.5 Chin type monostearate 1.5 Wax 0.5 Liquid paraffin 8.0 Squalane 7.0 Isocetyl octanoate 4.0 Refined jojoba oil 4.0 Dimethylsiloxane 0.3 Sorbitan monostearate 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 Origin plant, Taejung, Root culture extract 7.0 Purified water to 100

Production Example 2-5: Gel

According to the following prescription, it was prepared according to a conventional gel preparation method.

Raw material name Weight% (w / w) glycerin 4.0 Propylene glycol 4.0 ethanol 10 Polyoxyethylene hardened castor oil 0.1 Carboxy polymer 0.30 Triethanolamine 0.30 antiseptic Suitable amount Spices Suitable amount flash Suitable amount Origin plant, Taejung, Root culture extract 1.0 Purified water to 100

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the invention is not limited thereby. It will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (10)

Aster sphatholifolius Maxim.), or an extract thereof.
[Claim 7] The composition for external application for skin according to claim 1, wherein the skin improving agent is for skin aging.
The composition for external application for skin according to claim 1, wherein the skin-improving agent is for improving wrinkles.
The composition for external application for skin according to claim 1, wherein the skin improving agent is a skin barrier function-protecting or enhancing agent.
[Claim 7] The composition for external application for skin according to claim 1, wherein the skin improving agent is for skin elasticity enhancement.
Plant cell or adventitious root culture of Aster sphathulifolius Maxim., Or an extract thereof.
A plant skin or adventitious root culture of Aster sphathulifolius Maxim., Or an extract thereof.
A country that includes the following steps ( Aster sphathulifolius Maxim.) Method for preparing a skin external cytotoxic agent composition for skin improvement comprising a culture of a tubercle cell of a plant or an extract thereof:
(a) separating the leaf, stem, or embryo in the ovary, and culturing the plant cell or adventitious muscle; And
(b) a step of preparing an external skin application composition containing the cultured plant cell culture or adventitious root culture or extract thereof.
9. The method of claim 8, wherein step (a)
(i) isolating leaves or stem tissues of the transplanting plants and culturing them in a medium containing p-chlorophenoxyacetic acid and kinetin, or
(ii) separating the laver tissue in the ovary of the transplanting plant and culturing it in a medium containing p-chlorophenoxyacetic acid and zeatin, or
(iii) isolating leaves or stem tissues of the decoyed plants and culturing them in a medium containing p-chlorophenoxyacetic acid and indolebutyric acid to obtain an adventitious culture,
&Lt; / RTI &gt;
[9] The method according to claim 8, wherein the step (b) comprises: culturing the cultured Korean plant cell cultures or adventitious root cultures obtained in step (a), pulverizing and mixing them into purified water to prepare a composition, , Followed by sonication or hot water extraction to produce a composition.

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