KR20150090014A - Anti-aging Composition for Skin External Application Comprising Liriodendron tulipifera Placenta Culture Extracts - Google Patents

Abstract

The present invention relates to a skin external application composition including a Magnoliopsida placenta cell culture product or an extract thereof. More specifically, the present invention relates to an anti-aging skin external application composition including a Magnoliopsida placenta cell culture product or an extract thereof as an active ingredient. According to the present invention, the skin external application composition including a Magnoliopsida placenta cell culture product or an extract thereof is not toxic to a skin cell, has an excellent skin collagen synthesis ability, and has effects of reducing the size of a pore, whitening skin, inhibiting secretion of sebum, alleviating acne, moisturization, and anti-inflammation.

Description

[0001] The present invention relates to an anti-aging composition for external application for skin, which comprises a tulip tree cell culture extract,

More particularly, the present invention relates to an anti-aging dermatological composition containing Magnoliaceae procambium cell culture or its extract as an active ingredient. .

Sunlight is essential for our lives, but it has adverse effects on skin disorders such as skin cancer and aging, cataracts, and whole body immunity. Exposure to ultraviolet rays causes various skin changes such as inflammation and tissue damage, cancer development, and aging of the skin due to a biological reaction in the human skin. It is known that UVB is involved mainly in sunburn during skin reaction by ultraviolet rays, and the ability of erythema formation is about 1,000 times stronger than that of UVA.

Over time, the secretion of various hormones that regulate metabolism is reduced, and the function of immune cells and the activity of cells are lowered, so that the biosynthesis of immune proteins and biologic proteins necessary for the living body is reduced The skin becomes thinner (in the case of endogenous aging) and the elasticity is decreased by intrinsic aging which is caused by external aging and photoaging due to various pollutants and ultraviolet ray externally, Keratinocytes and melanocytes 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 protects the skin from external stimuli or forces, and it accounts for 90% of the dermis, so collagen reduction is closely related to skin and aging.

Cells that constitute the skin have decreased cell activity due to endogenous aging, photoaging, and imperfect signal transduction, resulting in increased biosynthesis of matrix metalloproteinases (MMPs), enzymes that degrade skin tissues, decreased collagen biosynthesis, And melanin production, which causes skin blackening, is known to increase. Therefore, it is possible to make skin thicker by proliferating skin cells or increasing the matrix material constituting the skin, a substance capable of inhibiting the biosynthesis of MMPs degrading collagen, which is a substrate substance constituting the skin, melanin biosynthesis Inhibiting substances and the like have been found to alleviate skin symptoms such as wrinkles, loss of elasticity, and blackening of the skin.

About 300,000 kinds of plants are known all over the world, and it has been found that the plants contain various kinds of bioactive active ingredients. These useful substances are secondary metabolites that are produced secondary to the normal metabolic processes of the cell and accumulate in specific locations in the cell. From the biochemical and ecological point of view, they protect the plant itself from microorganisms or external animals, Interest is growing in that it is an advantageous substance. The secondary metabolites produced by plants include alkaloids, flavonoids, carotenoids, glycosides, and terpenoids. These secondary metabolites have many effects such as moisturizing, skin calming and convergence, ultraviolet light blocking, whitening, exfoliation, skin aging prevention, antioxidation, skin wrinkle improvement, antibacterial, anti-inflammatory and anti-cancer effects.

On the other hand, a callus is an oily tissue in which a wound cell restores the ability to divide and protects against scratches when a wound is injured in a plant, and a special tissue mass produced by a method of culturing a tissue cut from the plant in a medium containing auxin And there is a growing interest in the use of such calluses and they are being studied in various fields.

In recent years, much research has been conducted on the development and application of functional foods using phytochemicals, such as phytochemicals. However, there has been a study on cosmetic compositions (Patent Document 1) concerning Magnoliae extracts on Magnolia, It is not at all.

Therefore, it is urgent to develop a functional cosmetic having various functions such as anti-aging, whitening, anti-acne, antibacterial, anti-inflammation, antioxidant,

Patent Document 1: Korean Patent No. 10-079551

While trying to produce a composition for external application for skin having anti-aging ability, the inventors of the present invention have found that the seedling of the Magnoliae plant is isolated and cultured through plant tissue culture, Growth and proliferation of the hair, and anti-aging, anti-inflammation and antioxidant effects including whitening and wrinkle-reducing effects, and the like.

Accordingly, an object of the present invention is to provide an anti-aging dermatologic external composition containing Magnolia thunbergii cell culture or its extract as an active ingredient.

It is still another object of the present invention to provide a method for producing a composition for external application for skin, which comprises the cultured Magnolia tephroblast cell or extract thereof as an active ingredient.

In order to achieve the above object, the present invention provides an anti-aging dermatologic external preparation composition comprising Magnoliaceae procambium cell culture or its extract as an active ingredient.

In the present invention, the magnolia is selected from the group consisting of Annoaceae, Degeneriaceae, Eupomatiaceae, Himantandraceae, Magnoliaceae, Schisandraceae, Lauraceae, ) Or Myristicaceae. ≪ / RTI >

In the present invention, the magnolia tree is selected from the group consisting of Annona squamosa, Eupomatia laurina, Magnolia, Magnolia grandiflora, Liriodendron tulipifera, Magnolia sieboldii, , Schizandra chinensis, and Cinnamomum japonicum.

In the present invention, the composition may be characterized by being used for anti-wrinkle or improving, anti-inflammatory, pore reduction, sebum secretion suppressing, acne improvement, skin moisturizing, whitening, or antibacterial.

(A) isolating and culturing the laver tissue in the ovary of a Magnolia plant; And (b) a step of preparing a culture containing the culture obtained in the step (a) or an extract thereof, and a method for producing an anti-aging dermatological composition containing the extract .

The granulomatous cell culture of Magnolia japonica according to the present invention or the composition for external application for skin containing the extract is excellent in the ability to synthesize skin collagen without toxicity to skin cells and has excellent anti-aging, skin moisturizing, anti- Sebum secretion suppression, acne improvement efficacy, has antibacterial power.

The present invention relates to an anti-aging dermatological composition containing Magnoliaceae procambium cell culture or its extract as an active ingredient, and a method for producing the same.

In one aspect, the present invention relates to an anti-aging dermatological composition containing Magnoliaceae procambium cell culture or its extract as an active ingredient. Specifically, the Magnoliaceae procambium cell 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.

In the present invention, the magnolia is selected from the group consisting of Annoaceae, Degeneriaceae, Eupomatiaceae, Himantandraceae, Magnoliaceae, Schisandraceae, Lauraceae, Myristicaceae, and the like.

In the present invention, the representative group belonging to Magnolia is Annona squamosa, Eupomatia laurina, Magnolia, Magnolia grandiflora, Liriodendron tulipifera, Magnolia sieboldii, Schizandra chinensis, Cinnamomum japonicum, more specifically Annona squamosa, Eupomatia laurina Magnolia kobus, Magnolia denudata, Magnolia grandiflora, Liriodendron tulipifera, Magnolia sieboldii, Magnolia liliiflora, Magnolia obovata, Magnolia denudata var purpurascens, Schisandra chinensis, And cinnamomum japonicum.

In addition, in the present invention, the magnolia thunbule cell culture or extract thereof may contain 0.1 to 10% by weight based on the total weight of the composition, and this ratio is merely a preferable range. For example, in the following examples, 1% , Or 2% by weight of the cells. However, it was confirmed that the cells had excellent efficacy even in 0.1%, 0.5%, 5%, and 10% cells, and the cell survival rate was not affected even at 10%. It is obvious to those skilled in the art that the skin has improved skin whitening, whitening, anti-inflammation, and wrinkle-improving functions even in the case of 20% or more than 30%.

In the present invention, it is preferable to use the Magnolia tephra cell culture (culture) itself, to use the culture by filtration, or to crush the culture, or to dry the culture to obtain powder, .

In the present invention, "extract" means an extract obtained by various known extraction methods such as pulverization of the Magnolia sieboldii tuberculosis cell culture extract, cold water extraction method, hot water extraction method and ethanol extraction method.

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 the like. 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, cold water (15-25 ° C), for example, is mixed with the above-mentioned placental tissue culture or its dried powder and extracted for 3 days to obtain 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. When extraction is carried out 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.

In the present invention, anti-aging means prevention and improvement of skin protection and skin condition, skin whitening, prevention or improvement of skin aging and wrinkles, contraction and shrinkage of pores, skin protection and alleviation of skin inflammation reaction, Barrier function improvement, skin irritation mitigation, skin cell proliferation and regeneration ability, antioxidant ability, and collagen synthesis promoting ability.

In the present invention, the term "comprising as an active ingredient" means a composition for external application for skin, which is capable of exhibiting a skin improving effect upon antiaging, for example, a composition for external application for skin, collagen synthesis associated with wrinkle- Quot; means an effective amount of such an amount as to exhibit whitening efficacy, skin moisturizing effect, pore contraction or iNOS inhibitory activity, sebum inhibition, antibacterial activity and the like.

Therefore, the composition for external application for skin according to the present invention can be applied for prevention of aging of skin and improvement / prevention of skin wrinkles because it has increased pro-collagen and collagen biosynthetic ability. In addition, whitening, shrinkage of pores, anti-inflammation through iNOS inhibitory activity, skin inflammation prevention, alleviation, alleviation, and treatment.

Among the various attractants, 100 ~ 300 M methyl jasmonic acid inducer treatment and high frequency treatment (high frequency treatment at 240 ~ 270 kHz for 3 ~ 5 times at 5 ~ 10 minute interval), 2 Repeating -4 weeks has better efficacy than if the inducer was not treated.

The efficacy of the Magnoliae thunbuocyte cell culture extract can be evaluated by comparing the extract of Magnolia thunbergii cell culture extract with that of other parts such as flowers, leaves, stem and root tissue-derived cell cultures, It has antioxidant, moisturizing, anti-wrinkle, whitening, pore shrinkage, sebum suppression, acne improvement, anti-bacterial effect and excellent polyphenol content and flavonoid content.

From this point of view, the composition for external application for skin according to the present invention includes compositions for improving whitening, improving wrinkles, preventing, anti-inflammation, skin moisturizing, pore shrinking, sebum secretion suppressing, acne improvement / prevention, and antibacterial.

In particular, in the case of a composition for antimicrobial use, it may be a composition having antimicrobial activity against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Candida yeast, and filamentous fungi as exemplified in the examples.

Plant tissue culture is carried out on small live plant tissue ( in in vitro ) and cultivating 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 .

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.

In another aspect, the present invention provides a method of producing a plant of the magnolia plant comprising the steps of: (a) And (b) preparing a composition containing the Magnoliaceae procambium cell culture or an extract thereof. The present invention also relates to a method for producing a composition for external application for skin, which comprises the extract of Magnolia thunbergii cell culture or extract thereof.

In the step (a), it is possible to select a suitable medium for culturing magnolia thrips cells. 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.

In the present invention, it is preferable that in the step (b), the composition containing the cultured magnolia thunbuocyte cell culture obtained through the cultivation in the step (a) is prepared as 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 the step (b), the cultured mammary goblet cells obtained in step (a) are dried, powdered and mixed with purified water to prepare a composition,

drying the Magnolia tephra cell culture obtained in step (a), pulverizing it, mixing it with purified water, and then preparing the composition by cold extraction, hot water extraction or ethanol extraction.

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

In the culture obtained in the step (a), 200 M methyl jasmonic acid was cultured in MS medium, or Magnoliales teat cell cultures were cultured in a high frequency cell culture incubator, and high frequency 240 ~ 270 kHz every 5 ~ 10 minutes at intervals of 3 ~ 5 times and it can be repeated for a week or two weeks. The MS medium can be incubated for 5 weeks with an air supply of 0.1 vvm in a 20 L balloon bioreactor at 25 and humidity 70% in a medium containing 1 mg / L 6-benzylaminopurine and 0.3 mg / L 2,4-D. The high-frequency wave, which is a type of physical attractant, is cultured in conjunction with a balloon type bioreactor (for example, patent application No. 10-2012-0127017) and treated at 5 kHz three times at 5 kHz with high frequency at 240 kHz for 5 days. It can be repeated for two weeks.

In this case, anti-inflammation, antioxidant, moisturizing, anti-wrinkle, whitening, pore contraction, sebum suppression, acne improvement, antibacterial and the like are more effective than those without treatment of the attractants.

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 hardened castor oil, ethanol, triethanolamine, etc., Preservatives, antioxidants, coloring agents, purified water, etc. can 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 preparing a composition for external application for skin containing Magnolia sieboldii cellulata extract according to the present invention is not limited to the above-described preparation method. Any person skilled in the art will be able to make some modifications It is also possible to prepare a composition for external application for skin containing a Magnolia sieboldii cell culture extract 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 cosmetic composition is manufactured from a cosmetic composition, the cosmetic composition of the emulsified formulation includes 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 as an agent for treating or preventing immune diseases, such as prevention of skin aging due to the ability to promote collagen synthesis, whitening due to inhibition of melanogenesis, mitigation / prevention of inflammation through inhibition of skin moisturizing iNOS activity, Inhibiting, preventing acne, and functioning cosmetics capable of antibacterial function.

In the present invention, the pharmaceutical composition has the iNOS activity inhibiting ability shown in the following examples, and can function as a pharmaceutical composition for the treatment and prevention of immune diseases.

Such a pharmaceutical composition may contain, in addition to the active ingredient Magnoliaceae procambium cell culture or extract thereof, a "pharmaceutically acceptable carrier ", which may be a diluent, a lubricant, a binder, a disintegrant, a sweetener, Preservatives, < / RTI > 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.

In particular, the following examples have confirmed the efficacy of Magnolia thunbergii cell culture extracts, but it will be apparent to those skilled in the art that such effects are also obtained with the culture itself, not with the extract.

According to the invention Magnolia tree ( Magnoliales ) Preparation of extracts by site

(1) Cell separation

1.1 Isolation of Magnoliales Tumor Cells

Magnoliales For the cultivation, 70% of the buds of Annona squamosa, Eupomatia laurina, Magnolia, Magnolias, Tulip trees, Rhododendrons, Ethanol and 0.5% sodium hypochlorite for 5 minutes and washed several times with sterile water. Carefully, the ovary wall was removed using tweezers, and the thyroid cells were separated and cultured in MS medium (Duchefa, Cat # M0256) containing 5% sucrose for 2 to 3 weeks.

1.2 Isolation of Magnoliales Flower Cells

Magnoliales For the cultivation, the petals of annona squamosa, Eupomatia laurina, magnolia, Magnolia, tulip, japonica, lilium, and balsam are represented by 70% ethanol And 0.5% sodium hypochlorite for 1 minute and washed several times with sterile water. Carefully cut with a tweezers and a sharp knife at once, and cut flower parts were dipped in MS medium (Duchefa, Cat # M0256) containing 5% sucrose and cultured for 2 to 3 weeks.

1.3 Isolation of Magnoliales Leaf Cells

Magnoliales For the cultivation, the leaves of Annona squamosa, Eupomatia laurina, Magnolia, Magnolia, Tulip tree, Prunus persicae, Schizandra chinensis, And 0.5% sodium hypochlorite for 1 minute and washed several times with sterile water. Carefully cut with a tweezers and a sharp knife at once, and the cut leaves were dipped in MS medium (Duchefa, Cat # M0256) containing 5% sucrose and cultured for 2 to 3 weeks.

1.4 Isolation of Magnoliales Stem Cells

Magnoliales For the cultivation, the leaves of Annona squamosa, Eupomatia laurina, Magnolia, Magnolia, Tulip tree, Prunus persicae, Schizandra chinensis, And 0.5% sodium hypochlorite for 1 minute and washed several times with sterile water. Carefully cut with a tweezers and a sharp knife at once, and the cut stems were streaked on MS medium (Duchefa, Cat # M0256) containing 5% sucrose and cultured for 2 to 3 weeks.

1.5 Isolation of Magnoliales Root Cells

Magnoliales For the cultivation, the leaves of Annona squamosa, Eupomatia laurina, Magnolia, Magnolia, Tulip tree, Prunus persicae, Schizandra chinensis, And 0.5% sodium hypochlorite for 1 minute and washed several times with sterile water. Carefully cut with a tweezers and a sharp knife at once, and the cut root was dipped in MS medium (Duchefa, Cat # M0256) containing 5% sucrose and cultured for 2 to 3 weeks.

(2) Separated Magnoliales Cultivation and mass production of turtles, flowers, leaves, stems and root cells

Aseptically isolated Magnoliales turtles, flowers, leaves, stems and roots were subcultured in MS medium containing 5% sucrose at intervals of 2 ~ 3 weeks. Afterwards, balloon bioreactor (Samsung Sci.) Was used to mass-produce liquid culture of the same composition except for agar at a temperature of 25 and a constant air supply of 0.1 vvm for 14 days.

(3) Magnoliales Manufacture of extracts of cultured turtle, flowers, leaves, stems and root cells

Magnoliales Magnoliales Typically, there are many species of plants, such as Annona squamosa, Eupomatia laurina, Magnolia, Magnolia, Tulipa, Zucchini, And root cells were harvested, and the water was sufficiently removed with a clean tissue, followed by drying in a dryer at 60 ° C for 2 days. 10 g of purified water was placed in a container, and the mixture was heated at 80 to 100 ° C. for 8 to 48 hours in a hot water distiller to obtain a hot water extract ≪ / RTI > After extraction, it is filtered with a mesh to remove solid matter, and Magnoliales are represented by Annona squamosa, Eupomatia laurina, Magnolia, Magnolia, Tulipa, Pinaceae, The cellulase extracts of P. japonica were prepared.

Experimental Example 1 Effect on survival of cells magnoliales (Magnoliales) cuts of the cell culture extract according to the invention Test

Human keratinocyte (HaCaT) was cultured in an ATCC (American Type Culture Collection) in order to examine the proliferative activity of the cell culture extract of Magnoliales according to the present invention. The cells were inoculated into a 24-well culture plate at a concentration of 1 × 10 ⁴ cells / ml. The medium was DMEM (Dubelco's Modified Eagle Medium, BRL, USA) containing 10% FBS. After culturing in DMEM containing 10% FBS for 48 hours and culturing by 25 to 30% of the surface area of the culture dish, the Magnoliales turmeric cell culture extract prepared in Example 1 was replaced with FBS-free DMEM containing 10% And cultured for another 24 hours. 50 μl of a solution of 2.5 mg / ml of 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT, Sigma M5655, USA) (DMSO, Sigma D2650, USA) was added to each well. The resulting solution was stirred for 20 minutes to dissolve the formazan crystals, and 100 μl of each was dissolved in 96 wells And the absorbance at 570 nm was measured by Enzyme-Linked Immunosorbent Assay (ELISA). The degree of proliferative activity of skin cells was calculated as a percentage based on the absorbance of the control group using pure water according to the following formula.

[Equation 1]

Cell viability effect (%) = (absorbance at the time of extract treatment / absorbance of control group) 100

Treatment concentration part Cell survival rate effect (%)
(Cell Viability) Control group 100 One % Annona squamosa Taurus 107 Flower 100 leaf 95 stem 98 Root 102 Eupomatia laurina Taurus 96 Flower 82 leaf 80 stem 75 Root 80 magnolia Taurus 90 Flower 70 leaf 75 stem 72 Root 78 Magnolia tree Taurus 110 Flower 99 leaf 100 stem 102 Root 100 Tulip tree Taurus 120 Flower 110 leaf 100 stem 120 Root 99 Shark tree Taurus 109 Flower 106 leaf 100 stem 99 Root 97 Cherry tree Taurus 90 Flower 87 leaf 89 stem 85 Root 80 Beech tree Taurus 115 Flower 105 leaf 102 stem 108 Root 110

Using the human keratinocyte, cell culture extracts of the representative plants of Magnoliales were treated to determine the suitable range for skin application, that is, the cell viability cell survival effect was confirmed. In the case of 1% concentration of cell culture extracts of Magnolia japonica, the contents of Annona squamosa, Eupomatia laurina, Magnolia, Magnolia, Tulipa, Rhododendron, The cell viability of the cells was not influenced by the cell culture extract.

Experimental Example  2: Identification and content test of extract components

It is an object of the present invention to provide a method for growing a plant belonging to the genus Magnoliales by cell culture and proliferating the plant and to provide a physical and chemical elicitor for increasing the content of polyphenolic compounds, Respectively.

Methods for increasing the content of carotenoids, flavonoids and phenolic compounds, secondary metabolites, by site of plant cells belonging to Magnoliales include

1. Incubate 200 M methyl jasmonic acid in MS medium.

2. Cell cultures of plant parts belonging to Magnoliales were cultivated in a high frequency cell incubator and treated with high frequency 240 or 270 kHz for 3 times at intervals of 5 minutes for one day and repeated for one week or two weeks.

The following experiments were carried out to confirm the total phenolic content and total flavonoid content of phenolic substances, flavonoids, and antioxidative effects of the major components of cell culture extracts of Magnoliales according to the present invention.

Total phenol content measurement

10 ml of distilled water was added to Extract 1, followed by addition of 2 ml of Folin-Ciocalteu phenol reagent (Sigma), followed by reaction at room temperature for 5 minutes. 2 ml of 20% sodium carbonate was added to the reaction mixture, and the mixture was incubated at room temperature for 1 hour. Then, the absorbance was measured at 680 nm using a microplate reader (UVT-06685, Thermo max, USA). At this time, gallic acid (Sigma, USA) was used as the indicator substance, and a calibration curve for the concentration of the indicator substance was prepared, and the absorbance value of each sample was substituted and shown in Table 2.

Total flavonoid content measurement

2% AlCl36H2O dissolved in the same amount of methanol was added to 1.5 ml of the cell culture extract of each Magnoliales portion. The mixture was incubated at room temperature for 10 minutes, and then incubated at 367 nm (UVT-06685, Thermo max, USA) The absorbance was measured. At this time, catechin (catechin, Sigma, USA) was used as the indicator material, and a calibration curve for the concentration of the indicator substance was prepared and the absorbance values of the respective samples were substituted and shown in Table 2.

process
density Incentive part Total phenol content
(mgg ^-1 )
(gallic acid equivalents) Total flavonoid content (mgg ^-1 )
(catechin equivalents) Control group One % Annona squamosa - Taurus 267 160 Flower 134 73 leaf 27 15 stem 16 9 Root 182 99 MeJA Taurus 401 218 Flower 214 116 leaf 134 73 stem 134 73 Root 267 145 RF Taurus 294 145 Flower 107 58 leaf 16 9 stem 16 9 Root 107 58 Eupomatia laurina - Taurus 331 180 Flower 27 15 leaf 53 29 stem 37 20 Root 11 6 MeJA Taurus 321 175 Flower 27 15 leaf 43 23 stem 27 15 Root 27 15 RF Taurus 428 233 Flower 134 73 leaf 150 81 stem 134 73 Root 134 73 magnolia - Taurus 321 175 Flower 27 15 leaf 32 17 stem 32 17 Root 43 23 MeJA Taurus 294 160 Flower 27 15 leaf 32 17 stem 32 17 Root 43 23 RF Taurus 283 154 Flower 32 17 leaf 27 15 stem 32 17 Root 32 17 Magnolia tree

- Taurus 380 207 Flower 64 35 leaf 32 17 stem 27 15 Root 203 111 MeJA Taurus 310 169 Flower 53 29 leaf 16 9 stem 16 9 Root 96 52 RF Taurus 481 262 Flower 160 87 leaf 139 76 stem 150 81 Root 310 169 Tulip tree - Taurus 385 209 Flower 294 160 leaf 176 96 stem 182 99 Root 374 204 MeJA Taurus 321 175 Flower 278 151 leaf 160 87 stem 150 81 Root 267 145 RF Taurus 481 262 Flower 315 172 leaf 283 154 stem 289 157 Root 374 204  Shark tree - Taurus 267 145 Flower 225 122 leaf 32 17 stem 21 12 Root 187 102 MeJA Taurus 160 87 Flower 118 64 leaf 32 17 stem 21 12 Root 53 29 RF Taurus 321 175 Flower 294 160 leaf 80 44 stem 75 41 Root 160 87 Cherry tree - Taurus 449 244 Flower 11 6 leaf 27 15 stem 32 17 Root 91 49 MeJA Taurus 428 233 Flower 16 9 leaf 27 15 stem 27 15 Root 53 29 RF Taurus 374 204 Flower 16 9 leaf 21 12 stem 21 12 Root 27 15  Beech tree - Taurus 385 209 Flower 331 180 leaf 32 17 stem 64 35 Root 246 134 MeJA Taurus 363 198 Flower 321 175 leaf 27 15 stem 53 29 Root 160 87 RF Taurus 470 256 Flower 417 227 leaf 86 47 stem 102 55 Root 299 163

The results of Table 2 show that the cell culture extracts induced by MeJA (methyl jasmonic acid) inducer in Annona squamosa showed high content of polyphenol and flavonoid, and in the treatment of radiofrequency inducer The cell culture extracts derived from Eupomatia laurina, Magnolia, Tulipa, Pinus densiflora, and Fagaceae showed high polyphenol contents and flavonoid contents. In addition, in the case of Magnolia and Schizandra chinensis, the highest polyphenol content and flavonoid content were found in the culture extract of untreated rabbit cell.

Experimental Example 3: Magnolia tree Tracheal cell culture extract Antioxidative effect ( ABTS Radical Scavenging Activity)

The antioxidant activity of ABTS radicals was measured by the method of van den Berg et al., Which is a method using ABTS free radicals produced by the reaction with potassium persulfate, Respectively.

1.0 mM AAPH (2,2'-azo-bis 2-amidinopropanediol hydrochloride) was mixed with 2.5 mM ABTS (2,2'-azino-bis-3-ethylbenzenothiazolin-6-sulfonic acid) dissolved in 100 mM PBS buffer. Lt; 0 > C for 12 minutes. 1% Concentration The absorbance of the extract of Example 1 (20 μl) and 980 μl ABTS solution was reacted at 37 ° C in a water bath for 10 minutes while measuring the absorbance at 734 nm. The positive control group was 0.1% Trolox. The ABTS radical scavenging activity was calculated according to the following equation (2). The results are shown in Table 3.

&Quot; (2) "

process
density Incentive part ABTS radical scavenging activity
(% of control) Control group 0 One % Annona squamosa - Taurus 40 Flower 25 leaf 5 stem 3 Root 34 MeJA Taurus 75 Flower 40 leaf 25 stem 25 Root 50 RF Taurus 50 Flower 20 leaf 3 stem 3 Root 20 Eupomatia laurina - Taurus 30 Flower 5 leaf 10 stem 7 Root 2 MeJA Taurus 60 Flower 5 leaf 8 stem 5 Root 5 RF Taurus 80 Flower 25 leaf 28 stem 25 Root 25 magnolia - Taurus 20 Flower 5 leaf 6 stem 6 Root 8 MeJA Taurus 55 Flower 5 leaf 6 stem 6 Root 8 RF Taurus 53 Flower 6 leaf 5 stem 6 Root 6

Magnolia tree - Taurus 45 Flower 12 leaf 6 stem 5 Root 38 MeJA Taurus 58 Flower 10 leaf 3 stem 3 Root 18 RF Taurus 90 Flower 30 leaf 26 stem 28 Root 58   Tulip tree - Taurus 45 Flower 55 leaf 33 stem 34 Root 30 MeJA Taurus 60 Flower 52 leaf 30 stem 28 Root 50 RF Taurus 90 Flower 59 leaf 53 stem 54 Root 70 Shark tree - Taurus 40 Flower 42 leaf 6 stem 4 Root 35 MeJA Taurus 30 Flower 22 leaf 6 stem 4 Root 10 RF Taurus 60 Flower 55 leaf 15 stem 14 Root 30  Cherry tree - Taurus 25 Flower 2 leaf 5 stem 6 Root 17 MeJA Taurus 80 Flower 3 leaf 5 stem 5 Root 10 RF Taurus 70 Flower 3 leaf 4 stem 4 Root 5 Beech tree - Taurus 50 Flower 52 leaf 6 stem 12 Root 46 MeJA Taurus 68 Flower 60 leaf 5 stem 10 Root 30 RF Taurus 88 Flower 78 leaf 16 stem 19 Root 56

As can be seen from the above Table 3, it was confirmed that the extract of the tubercle cell of the 1% concentration of the magnolia tree of the present invention showed the most excellent antioxidative activity than the cell culture extract derived from other sites.

Experimental Example  4: B16F10 malanoma cell Using melanogenesis  Inhibition measurement

Melanin-producing cells, B16F10, were cultured in DMEM medium supplemented with 10% FBS and 1% antibiotics at 37 ° C, 5%, and CO2. B16F10 cells were suspended in DMEM medium containing 10% FBS at a concentration of 1.5 × 10 ³ cells / well in order to examine the change in melanin production by 1% Magnolia thunbergii cell culture extract. Suspension cells (5 ml) are placed in a tissue culture flask and allowed to attach for 1 day. After adherence, a test sample was added and incubated at 37 ° C for 5 days in 5% CO2. The cultured B16F10 cells were washed with phosphate buffered saline (PBS) and trypsinized. After collection of cells in corning tube into the 1 × 10 ⁶ cell / per 1ml 1N NaOH solution and then dissolving the cells, by measuring the absorbance at 490nm with microplate reader it was determined melanogenesis inhibition.

&Quot; (3) "

Inhibition of melanogenesis (%) = [1 - (Absample / Abcontrol)] 100

Treatment concentration part Melanin Contents
(% of control) Control group 100 One % Annona squamosa Taurus 91 Flower 100 leaf 95 stem 100 Root 98 Eupomatia laurina Taurus 86 Flower 100 leaf 101 stem 100 Root 100 magnolia Taurus 75 Flower 100 leaf 99 stem 95 Root 98 Magnolia tree Taurus 80 Flower 100 leaf 98 stem 100 Root 100 Tulip tree Taurus 75 Flower 90 leaf 100 stem 85 Root 95 Shark tree Taurus 83 Flower 95 leaf 98 stem 100 Root 100 Cherry tree Taurus 80 Flower 98 leaf 97 stem 100 Root 100  Beech tree Taurus 85 Flower 95 leaf 98 stem 92 Root 90

In order to investigate the effect of cell culture extract of Magnolia sieboldii plant on melanin synthesis of B16 / F10 melanoma cells, total melanin level was measured after culturing for 3 days at a concentration of 1%. As a result, it was confirmed that the whitening effect of 1% magnolia seed plant cell culture extract decreased melanin synthesis only in the thyroid gland, and the whitening effect in the remaining area was insufficient.

In addition, in the case of the cells cultured by 200 M methyl jasmonic acid inducer treatment, the extract of the thyroid cell cultures showed a decrease of melanin synthesis about twice as much as that of the untreated case, The cell culture extracts showed a decrease in melanin synthesis of 1.2 times.

Experimental Example  5: Procollagen  Synthetic enhancement test

Human Fibroblast was cultured in DMEM (Dulbecco ' s Modified) containing 10% FBS, Penicillin (50 U / ml), Streptomycin (50 / ml) in a cell incubator maintained at a constant humidity in a 5% Eagle's Medium, Gibco, USA), and the cells were seeded at a density of 1 × 10 ⁵ / ml in a 24-well plate (500 μl) for 24 hours. The medium containing the control (DMEM medium) and cell culture extracts of the magnolia tree plant parts obtained in Example 1 at a concentration of 1% was added and cultured in a 5% CO2 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 < ⁵ > cells, calculated according to the following formula, and the results are shown in Table 5.

&Quot; (4) "

Treatment concentration part Increase in collagen biosynthesis
(% of control) Control group 100 One % Annona squamosa Taurus 117 Flower 110 leaf 105 stem 108 Root 112 Eupomatia laurina Taurus 106 Flower 92 leaf 90 stem 85 Root 78 magnolia Taurus 140 Flower 80 leaf 85 stem 82 Root 88 Magnolia tree Taurus 120 Flower 109 leaf 110 stem 112 Root 110 Tulip tree Taurus 130 Flower 120 leaf 110 stem 130 Root 109 Shark tree Taurus 119 Flower 116 leaf 110 stem 109 Root 107 Cherry tree Taurus 100 Flower 97 leaf 109 stem 75 Root 70 Beech tree Taurus 125 Flower 115 leaf 112 stem 118 Root 120

As shown in Table 5, it was confirmed that the amount of Procollagen biosynthesis was greatly increased by treating the extract of tuber cell cultures among the magnolia plant parts at a concentration of 1% in Example 1, It can be seen that the collagen synthesis enhancing effect is shown by each part of the fish.

In addition, in the case of the cells cultured by the treatment with 200 M methyl jasmonic acid inducer, the amount of procollagen biosynthesis increased 2.5 times as compared with that in the case of untreated cells, The culture extract showed an increase in the amount of Procollagen biosynthesis about 1.2 times.

Experimental Example  6: iNOS  Antiinflammatory effect experiment by active inhibition

The amount of nitric oxide (NO) produced from the Raw 264.7 cell line was determined using the Griess reagent as the NO 2 -form present in the cell culture. In order to measure inhibition of NO production in 1 g LPS-activated Raw 264.7 cells, 1% concentration of the cell culture extract of Magnolia sieboldii was treated with the cell culture extract for 24 hours and the Griess reagent was added to the cell culture supernatant 100 100 μl of Griess reagent (1% sulfanilamide in 5% phosphoric acid, 1% -naphthylamide in H2O) were mixed and reacted on 96-well plates for 10 minutes and absorbance was measured at 540 nm. The concentration of NO2- was obtained by measuring the absorbance by diluting sodium nitrate.

Treatment concentration part Cell Viability (%) NO Production (M) LPS treatment
(-) - Control group - 100 1.5 Control group 65 3.5




LPS treatment
(+)



One %  Annona squamosa Taurus 107 2.5 Flower 100 2.9 leaf 95 2.8 stem 98 2.7 Root 102 2.3  Eupomatia laurina Taurus 96 2.0 Flower 82 3.5 leaf 80 3.4 stem 75 3.5 Root 68 3.5  magnolia Taurus 101 1.9 Flower 70 3.0 leaf 75 3.2 stem 72 3.2 Root 78 3.3  Magnolia tree Taurus 110 2.0 Flower 99 3.5 leaf 100 3.4 stem 102 2.8 Root 100 3.5 Tulip tree Taurus 120 1.5 Flower 110 2.3 leaf 100 2.0 stem 110 2.0 Root 99 2.4 Shark tree Taurus 109 2.7 Flower 106 3.0 leaf 100 3.5 stem 99 3.4 Root 97 3.5 Cherry tree Taurus 90 1.9 Flower 87 3.4 leaf 89 3.3 stem 65 3.5 Root 60 3.4 Beech tree Taurus 105 1.8 Flower 105 2.0 leaf 102 2.3 stem 100 2.1 Root 100 2.3

As shown in Table 6, the activity of iNOS inhibited the production of NO compared with the control group by treatment of the extract of the tubercle cell culture of 1% Magnolia plant, and showed the most excellent inflammation inhibitory effect.

In addition, the cells were cultured in a high frequency cell incubator for 3 days at a frequency of 240 to 270 kHz for 5 days and cultured in a high frequency cell culture incubator. INOS activity was 1.8 times more than that of untreated cells, whereas other cell culture extracts were ineffective.

Experimental Example  7: skin moisturizing effect experiment

1. Annona squamosa

Raw material name Weight% (w / w) compare
Produce
Example 1 Produce
Example 1-1 Produce
Example 1-2 Produce
Example 1-3 Produce
Example 1-4 Produce
Example 1-5 Cetostearyl alcohol 1.0 1.0 1.0 1.0 1.0 1.0 Self emulsifying monostearic acid 1.0 1.0 1.0 1.0 1.0 1.0 Wax 0.5 0.5 0.5 0.5 0.5 0.5 Squalane 5.0 5.0 5.0 5.0 5.0 5.0 Isocetyl octanoate 3.0 3.0 3.0 3.0 3.0 3.0 Dimethylsiloxane 0.3 0.3 0.3 0.3 0.3 0.3 Sorbitan monostearate 0.5 0.5 0.5 0.5 0.5 0.5 Polyethylene glycol monostearate 8.0 8.0 8.0 8.0 8.0 8.0 glycerin 4.0 4.0 4.0 4.0 4.0 4.0 Propylene glycol 0.2 0.2 0.2 0.2 0.2 0.2 Carboxy polymer 0.22 0.22 0.22 0.22 0.22 0.22 Triethanolamine 0.25 0.25 0.25 0.25 0.25 0.25 antiseptic Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Spices Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount flash Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Annona squamosa Thyroid cell culture extract - 2 - - - - Annona squamosa flower cell culture extract - - 2 - - - Leaf cell culture extract of Annona squamosa - - - 2 - - Annona squamosa stem cell culture extract - - - - 2 - Root cell culture extract of Annona squamosa - - - - - 2 Purified water to 100 to 100 to 100 to 100 to 100 to 100

2. Eupomatia laurina (Eupomatia laurina)

Raw material name Weight% (w / w) compare
Produce
Example 2 Produce
Example 2-1 Produce
Example 2-2 Produce
Example 2-3 Produce
Example 2-4 Produce
Example 2-5 Cetostearyl alcohol 1.0 1.0 1.0 1.0 1.0 1.0 Self emulsifying monostearic acid 1.0 1.0 1.0 1.0 1.0 1.0 Wax 0.5 0.5 0.5 0.5 0.5 0.5 Squalane 5.0 5.0 5.0 5.0 5.0 5.0 Isocetyl octanoate 3.0 3.0 3.0 3.0 3.0 3.0 Dimethylsiloxane 0.3 0.3 0.3 0.3 0.3 0.3 Sorbitan monostearate 0.5 0.5 0.5 0.5 0.5 0.5 Polyethylene glycol monostearate 8.0 8.0 8.0 8.0 8.0 8.0 glycerin 4.0 4.0 4.0 4.0 4.0 4.0 Propylene glycol 0.2 0.2 0.2 0.2 0.2 0.2 Carboxy polymer 0.22 0.22 0.22 0.22 0.22 0.22 Triethanolamine 0.25 0.25 0.25 0.25 0.25 0.25 antiseptic Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Spices Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount flash Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Eupomatia laurina Thyroid cell culture extract - 2 - - - - Eupomatia laurina flower cell culture extract - - 2 - - - Leaf cell culture extract of Eupomatia laurina - - - 2 - - Eupomatia laurina stem cell culture extract - - - - 2 - Root cell culture extract of Eupomatia laurina - - - - - 2 Purified water to 100 to 100 to 100 to 100 to 100 to 100

3. Magnolia

Raw material name Weight% (w / w) compare
Produce
Example 3 Produce
Example 3-1 Produce
Example 3-2 Produce
Example 3-3 Produce
Example 3-4 Produce
Example 3-5 Cetostearyl alcohol 1.0 1.0 1.0 1.0 1.0 1.0 Self emulsifying monostearic acid 1.0 1.0 1.0 1.0 1.0 1.0 Wax 0.5 0.5 0.5 0.5 0.5 0.5 Squalane 5.0 5.0 5.0 5.0 5.0 5.0 Isocetyl octanoate 3.0 3.0 3.0 3.0 3.0 3.0 Dimethylsiloxane 0.3 0.3 0.3 0.3 0.3 0.3 Sorbitan monostearate 0.5 0.5 0.5 0.5 0.5 0.5 Polyethylene glycol monostearate 8.0 8.0 8.0 8.0 8.0 8.0 glycerin 4.0 4.0 4.0 4.0 4.0 4.0 Propylene glycol 0.2 0.2 0.2 0.2 0.2 0.2 Carboxy polymer 0.22 0.22 0.22 0.22 0.22 0.22 Triethanolamine 0.25 0.25 0.25 0.25 0.25 0.25 antiseptic Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Spices Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount flash Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Magnolia thunbuocyte culture extract - 2 - - - - Magnolia flower cell culture extract - - 2 - - - Magnolia leaf cell culture extract - - - 2 - - Magnolia stem cell culture extract - - - - 2 - Magnolia root cell culture extract - - - - - 2 Purified water to 100 to 100 to 100 to 100 to 100 to 100

4. Magnolia tree

Raw material name Weight% (w / w) compare
Produce
Example 4 Produce
Example 4-1 Produce
Example 4-2 Produce
Example 4-3 Produce
Example 4-4 Produce
Example 4-5 Cetostearyl alcohol 1.0 1.0 1.0 1.0 1.0 1.0 Self emulsifying monostearic acid 1.0 1.0 1.0 1.0 1.0 1.0 Wax 0.5 0.5 0.5 0.5 0.5 0.5 Squalane 5.0 5.0 5.0 5.0 5.0 5.0 Isocetyl octanoate 3.0 3.0 3.0 3.0 3.0 3.0 Dimethylsiloxane 0.3 0.3 0.3 0.3 0.3 0.3 Sorbitan monostearate 0.5 0.5 0.5 0.5 0.5 0.5 Polyethylene glycol monostearate 8.0 8.0 8.0 8.0 8.0 8.0 glycerin 4.0 4.0 4.0 4.0 4.0 4.0 Propylene glycol 0.2 0.2 0.2 0.2 0.2 0.2 Carboxy polymer 0.22 0.22 0.22 0.22 0.22 0.22 Triethanolamine 0.25 0.25 0.25 0.25 0.25 0.25 antiseptic Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Spices Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount flash Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Extract - 2 - - - - Magnolia flower cell culture extract - - 2 - - - Leaf cell culture extract - - - 2 - - Magnolia stems cell culture extract - - - - 2 - Root cell culture extract of Magnolia - - - - - 2 Purified water to 100 to 100 to 100 to 100 to 100 to 100

5. Tulip tree

Raw material name Weight% (w / w) compare
Produce
Example 5 Produce
Example 5-1 Produce
Example 5-2 Produce
Example 5-3 Produce
Example 5-4 Produce
Example 5-5 Cetostearyl alcohol 1.0 1.0 1.0 1.0 1.0 1.0 Self emulsifying monostearic acid 1.0 1.0 1.0 1.0 1.0 1.0 Wax 0.5 0.5 0.5 0.5 0.5 0.5 Squalane 5.0 5.0 5.0 5.0 5.0 5.0 Isocetyl octanoate 3.0 3.0 3.0 3.0 3.0 3.0 Dimethylsiloxane 0.3 0.3 0.3 0.3 0.3 0.3 Sorbitan monostearate 0.5 0.5 0.5 0.5 0.5 0.5 Polyethylene glycol monostearate 8.0 8.0 8.0 8.0 8.0 8.0 glycerin 4.0 4.0 4.0 4.0 4.0 4.0 Propylene glycol 0.2 0.2 0.2 0.2 0.2 0.2 Carboxy polymer 0.22 0.22 0.22 0.22 0.22 0.22 Triethanolamine 0.25 0.25 0.25 0.25 0.25 0.25 antiseptic Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Spices Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount flash Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Tulip tree cell culture extract - 2 - - - - Tulip tree flower cell culture extract - - 2 - - - Tulip leaf cell culture extract - - - 2 - - Tulip tree stem cell culture extract - - - - 2 - Tulip root cell culture extract - - - - - 2 Purified water to 100 to 100 to 100 to 100 to 100 to 100

6. Fir tree

Raw material name Weight% (w / w) compare
Produce
Example 6 Produce
Example 6-1 Produce
Example 6-2 Produce
Example 6-3 Produce
Example 6-4 Produce
Example 6-5 Cetostearyl alcohol 1.0 1.0 1.0 1.0 1.0 1.0 Self emulsifying monostearic acid 1.0 1.0 1.0 1.0 1.0 1.0 Wax 0.5 0.5 0.5 0.5 0.5 0.5 Squalane 5.0 5.0 5.0 5.0 5.0 5.0 Isocetyl octanoate 3.0 3.0 3.0 3.0 3.0 3.0 Dimethylsiloxane 0.3 0.3 0.3 0.3 0.3 0.3 Sorbitan monostearate 0.5 0.5 0.5 0.5 0.5 0.5 Polyethylene glycol monostearate 8.0 8.0 8.0 8.0 8.0 8.0 glycerin 4.0 4.0 4.0 4.0 4.0 4.0 Propylene glycol 0.2 0.2 0.2 0.2 0.2 0.2 Carboxy polymer 0.22 0.22 0.22 0.22 0.22 0.22 Triethanolamine 0.25 0.25 0.25 0.25 0.25 0.25 antiseptic Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Spices Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount flash Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Tracheal cell culture extract - 2 - - - - Seaweed flower cell culture extract - - 2 - - - Cell culture extract - - - 2 - - Stem cell culture extract - - - - 2 - Root cell culture extract - - - - - 2 Purified water to 100 to 100 to 100 to 100 to 100 to 100

7. Schizophyllum

Raw material name Weight% (w / w) compare
Produce
Example 7 Produce
Example 7-1 Produce
Example 7-2 Produce
Example 7-3 Produce
Example 7-4 Produce
Example 7-5 Cetostearyl alcohol 1.0 1.0 1.0 1.0 1.0 1.0 Self emulsifying monostearic acid 1.0 1.0 1.0 1.0 1.0 1.0 Wax 0.5 0.5 0.5 0.5 0.5 0.5 Squalane 5.0 5.0 5.0 5.0 5.0 5.0 Isocetyl octanoate 3.0 3.0 3.0 3.0 3.0 3.0 Dimethylsiloxane 0.3 0.3 0.3 0.3 0.3 0.3 Sorbitan monostearate 0.5 0.5 0.5 0.5 0.5 0.5 Polyethylene glycol monostearate 8.0 8.0 8.0 8.0 8.0 8.0 glycerin 4.0 4.0 4.0 4.0 4.0 4.0 Propylene glycol 0.2 0.2 0.2 0.2 0.2 0.2 Carboxy polymer 0.22 0.22 0.22 0.22 0.22 0.22 Triethanolamine 0.25 0.25 0.25 0.25 0.25 0.25 antiseptic Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Spices Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount flash Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Tumor cell culture extract of Schizandra chinensis - 2 - - - - Omija flower cell culture extract - - 2 - - - Omija leaf cell culture extract - - - 2 - - Omija stem cell culture extract - - - - 2 - Root cell culture extract of Schizandra chinensis - - - - - 2 Purified water to 100 to 100 to 100 to 100 to 100 to 100

8. Fruit trees

Raw material name Weight% (w / w) compare
Produce
Example 8 Produce
Example 8-1 Produce
Example 8-2 Produce
Example 8-3 Produce
Example 8-4 Produce
Example 8-5 Cetostearyl alcohol 1.0 1.0 1.0 1.0 1.0 1.0 Self emulsifying monostearic acid 1.0 1.0 1.0 1.0 1.0 1.0 Wax 0.5 0.5 0.5 0.5 0.5 0.5 Squalane 5.0 5.0 5.0 5.0 5.0 5.0 Isocetyl octanoate 3.0 3.0 3.0 3.0 3.0 3.0 Dimethylsiloxane 0.3 0.3 0.3 0.3 0.3 0.3 Sorbitan monostearate 0.5 0.5 0.5 0.5 0.5 0.5 Polyethylene glycol monostearate 8.0 8.0 8.0 8.0 8.0 8.0 glycerin 4.0 4.0 4.0 4.0 4.0 4.0 Propylene glycol 0.2 0.2 0.2 0.2 0.2 0.2 Carboxy polymer 0.22 0.22 0.22 0.22 0.22 0.22 Triethanolamine 0.25 0.25 0.25 0.25 0.25 0.25 antiseptic Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Spices Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount flash Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Tangerine cell culture extract - 2 - - - - Cellulase extract - - 2 - - - Cell culture extract - - - 2 - - Mesh stem cell culture extract - - - - 2 - Root cell culture extract - - - - - 2 Purified water to 100 to 100 to 100 to 100 to 100 to 100

In order to examine the skin moisturizing ability during one application, the cosmetic composition was applied to the inside of the face and anterior chamber to 50 persons who were in the 30s to 40s who were thought to be dry skin or dry skin, The skin moisture was measured using a corneometer (CM820 courage Khazaka electronic GmbH, Germany). The coneometer measures the amount of moisture present in the epidermis of the skin by measuring the degree of ion of water using a sensor and numerically expressing the amount of water to calculate the amount of moisture, and the measurement method is as follows.

1) A coneometer probe was placed on the skin area to be measured.

2) When the probe is pressed onto the skin, the electrical conductivity of the skin is quantified through the sensor and displayed on the screen.

3) Measurement was repeated with different measurement sites.

4) After the measurement, the sensor was wiped with a tissue paper such as a kim wipe and then measured again.

The moisture content of the skin was measured using a coneometer at a constant temperature and humidity (40% humidity) under the constant temperature and humidity conditions before initiation of the application, and the changes were measured after 12 hours and 24 hours. The results are shown in Table 15 below.

Skin moisture (%) sample Before application After 12 hours After 24 hours Comparative Preparation Example 1 30.9 38.5 31.4 Production Example 1-1 30.5 41.4 37.7 Production Example 1-2 30.3 39.9 36.3 Production Example 1-3 29.9 39.9 35.9 Production Example 1-4 30.2 40.3 36.5 Production Example 1-5 30.5 41.5 35.6 Comparative Production Example 2 30.7 38.5 31.4 Production Example 2-1 30.4 45.1 43.3 Production example 2-2 30.8 40.1 37.9 Production Example 2-3 30.1 41.2 37.8 Production example 2-4 30.2 41.5 37.5 Production example 2-5 30.3 40.3 37.1 Comparative Production Example 3 30.9 38.4 31.2 Production example 3-1 30.5 46.2 42.2 Production example 3-2 30.3 43.1 39.1 Production Example 3-3 29.9 42.8 38.2 Production example 3-4 30.2 42.1 38.7 Production Example 3-5 30.5 42.5 37.9 Comparative Production Example 4 30.6 38.6 31.6 Production Example 4-1 30.1 44.1 40.7 Production example 4-2 30.1 42.1 36.3 Production Example 4-3 30.1 41.9 36.9 Production Example 4-4 30.2 40.9 36.5 Production Example 4-5 30.3 40.5 37.7 Comparative Preparation Example 5 30.6 38.2 31.2 Production Example 5-1 30.2 48.9 44.5 Production Example 5-2 30.3 43.2 42.1 Production example 5-3 30.9 45.1 41.2 Production Example 5-4 30.2 44.9 41.9 Production Example 5-5 30.5 45.0 41.1 Comparative Preparation Example 6 30.7 38.4 31.5 Production example 6-1 30.4 45.2 43.1 Production Example 6-2 30.8 44.4 42.9 Production example 6-3 30.1 43.2 41.9 Production Example 6-4 30.2 44.5 40.8 Production Example 6-5 30.3 43.4 41.5 Comparative Preparation Example 7 30.9 38.3 31.8 Production Example 7-1 30.5 46.0 41.0 Production Example 7-2 30.3 41.2 42.1 Production Example 7-3 30.1 41.3 40.0 Production Example 7-4 30.2 42.5 39.9 Production Example 7-5 30.5 42.6 41.1 Comparative Preparation Example 8 30.7 38.2 31.0 Production Example 8-1 30.4 49.0 44.0 Production Example 8-2 30.8 43.2 42.1 Production example 8-3 30.1 46.1 41.2 Production Example 8-4 30.2 45.9 41.9 Production Example 8-5 30.3 46.0 41.1

From the above results, it was found that when Comparative Production Examples 1 to 8 were applied, the amount of skin moisture was increased to some extent after 12 hours from application, but returned to the state before application after 24 hours. On the other hand, extracts of cell culture extracts of parts of 1% concentration Magnolia japonica (Annona squamosa), Eupomatia laurina, Magnolia, Magnolia, Tulip tree, It was confirmed that the moisturizing effect lasts more than 24 hours and the skin moisture content increasing effect is also higher than that of Comparative Production Examples 1 to 8. When the skin external application compositions of Examples 1-1 to 8-5 are applied to skin,

In addition, 200 μM methyl jasmonic acid inducer treatment and high frequency cell culture were performed for 3 days at 240-270 kHz for 5 days at a frequency of 240-270 kHz, In the case of the cells, the extract of the tuber cell cultures showed a 1.5 times better skin moisture content than the untreated control, and the other cell culture extracts showed a 1.1 times increase in skin moisture content.

Experimental Example  8: Pore Shrinkage Effect Test In vitro Test )

In order to measure the pore-shrinking effect of the composition of the present invention, the composition prepared in the above example was tested for pore-shrinking effect using hemoglobin as a protein to replace skin. A hemoglobin solution (0.05 g / 50 ml) was prepared with 0.9% Phosphate Buffer Saline (0.1 mM, pH 7.4), 2 ml of the composition obtained in the above Example was added to 2 ml of the hemoglobin solution, The mixture was centrifuged at 3,500 rpm for 10 minutes, and 2 ml of purified water was added to 1 ml of the supernatant. The measurement was carried out at 407 nm in a UV-Visible spectrum. As a control, 2 ml of PBS (Phosphate Buffer Saline) was added in place of the composition of the above Example. In the case of the experimental group containing various concentrations of the composition, the shrinkage effect was measured by measuring the amount of hemoglobin settled. The higher the sedimentation rate (%) of hemoglobin was, the better the shrinkage effect of pores was judged.

The shrinkage effect was evaluated by measuring the sedimentation amount of hemoglobin. The higher the sedimentation rate (%) of hemoglobin was, the better the shrinkage effect was judged. As shown in the results of Table 16, cell culture extracts of 1% magnolia plant part showed significant difference and showed pore shrinking effect.

In addition, 200 μM methyl jasmonic acid inducer treatment and high frequency cell culture were performed for 3 days at 240-270 kHz for 5 days at a frequency of 240-270 kHz, In the case of the cells, the extract of Toc cell culture showed 2.2 times better pore shrinkage than that of untreated, and the other cell culture extract showed 1.1 times increase in pore shrinkage.

Treatment concentration part Precipitation of hemoglobin
(% of control) Control group 2.9 One % Annona squamosa, Taurus 24.8 Flower 15.3 leaf 11.1 stem 13.1 Root 7.9 Eupomatia laurina Taurus 19.3 Flower 12.3 leaf 15.7 stem 13.0 Root 11.1 magnolia Taurus 5.1 Flower 2.9 leaf 3.5 stem 3.0 Root 2.5 Magnolia tree Taurus 28.0 Flower 20.1 leaf 17.9 stem 15.5 Root 13.1 Tulip tree Taurus 33.0 Flower 30.4 leaf 25.8 stem 21.1 Root 25.8 Shark tree Taurus 20.3 Flower 15.5 leaf 15.2 stem 8.8 Root 7.9 Cherry tree Taurus 20.3 Flower 10.3 leaf 8.9 stem 5.3 Root 7.9 Beech tree Taurus 32.4 Flower 16.2 leaf 8.9 stem 8.0 Root 5.5

Experimental Example  9: Anti-sebum effect experiment

Ten subjects were tested for secretion of sebum on skin using a skin oil analyzer (Sebum meter SM810) 4 weeks after using the cell culture extract of the 1% Magnolia sieboldii plant part of the present invention. Experiments were performed at 22 ± 2 ° C and 40 ± 5% humidity. In the case of oily skin, the measured value is 220 / / cm2 or more, and in case of normal skin, it is 100 to 220 / / cm2. As a control, purified water was used. The results of the experiment are shown in Table 17.

Treatment concentration part Sebum suppression effect 0 day (before use) 28 days (after use) Control group 230 225 One % Annona squamosa Taurus 231 175 Flower 229 200 leaf 230 205 stem 232 201 Root 230 210 Eupomatia laurina Taurus 229 200 Flower 231 210 leaf 229 215 stem 230 195 Root 232 199  magnolia Taurus 230 210 Flower 231 205 leaf 232 205 stem 231 210 Root 231 190  Magnolia tree Taurus 230 190 Flower 232 195 leaf 231 205 stem 231 210 Root 230 205 Tulip tree Taurus 230 155 Flower 231 175 leaf 231 180 stem 230 190 Root 231 185 Shark tree Taurus 231 175 Flower 230 178 leaf 230 180 stem 231 190 Root 230 191 Cherry tree Taurus 230 179 Flower 231 199 leaf 231 205 stem 230 200 Root 229 210   Beech tree Taurus 229 170 Flower 231 185 leaf 230 190 stem 229 190 Root 230 195

As shown in the results of Table 17 above, the extract of the 1% Magnolia thunbergii cell culture showed the best inhibitory effect on sebum secretion compared with the control group.

In addition, 200 μM methyl jasmonic acid inducer treatment and high frequency cell culture were performed for 3 days at 240-270 kHz for 5 days at a frequency of 240-270 kHz, In the case of the cells, the extract of Tacrine cell culture showed a 2.5 times more inhibitory effect on sebum secretion than that of untreated control, and the other cell culture extract showed 1.3 times more inhibitory effect on sebum secretion.

Experimental Example  10: Antimicrobial activity measurement

The antimicrobial test was carried out by the solid culture dilution method by the test for the antimicrobial activity of the extract of the 1% magnolia thunbergii plant of the present invention. Staphylococcus aureus KCTC 6910, Gram-negative bacteria Pseudomonas aeruginosa KCTC 1637, E. coli KCTC 1039, and Candida yeast, respectively, were purchased from Korea Research Institute of Bioscience and Biotechnology. albicans KCTC 7965) and aspergillus (Aspergillus nigerKCTC 6910).

Agar Serial Dilution Method

Tritic soy broth was used as the fungi, and the broth was inoculated on the medium and pre-cultured for 24 hours in a 37 ° C shaking incubator. Potato dextrose culture medium was used for culture of yeast, and the bacteria were inoculated on the medium and cultured for 2 days in a 25 ° C shaking incubator. Potato dextrose agar medium was used for culture of molds, and the bacteria were inoculated on the medium and pre-cultured for 7 days in a 25 ° C incubator.

More specifically, in the case of bacteria, bacteria are inoculated on a tryptic soy broth and cultured at 37 ° C for 24 hours. In the case of yeast, bacteria are inoculated on a potato dextrose medium and cultured for 2 days at 25 ° C. The filamentous fungus was inoculated on a potato dextrose agar medium and pre-cultured for 7 days in a 25 ° C. incubator. Spores of filamentous fungus formed on the surface of the medium were recovered by using a smear rod and diluted in sterilized saline. 2 ml of each of the extracts diluted to the appropriate concentration in 5% DMSO (dimethylsulfoxide) physiological saline solution was added to the sterilized Patriedish by the sample and the experimental species, and the control was added to 2 ml of each sample diluted to a proper concentration in 5% physiological saline solution , And 2 ml of a 5% DMSO physiological saline solution was added to the control. Then, 18 ml of a tryptic soya agar medium and a potato dextrose agar medium, which had been sterilized in each Patridish and cooled to 48 ° C, were added, stirred and allowed to stand and solidify. Each of the pre-cultivated test bacteria was applied to each petri dish at a final concentration of about 1 × 10 ⁶ CFU / ml for bacteria, 1 × 10 ⁵ CFU / ml for yeast, about 1 × 10 ⁵ CFU / ⁴ CFU / ml in each petri dish. Each petri dish was incubated at 37 ° C for 24 hours, and the yeast was incubated at 25 ° C for 3 days. The molds were cultured in a 25 ° C incubator for 7 days, and then cloin formation was observed in each compartment. The minimum sample concentration was defined as the minimum inhibitory concentration (MIC), and the results are shown in Table 18. At this time, the minimum inhibitory concentration means that the smaller the value, the higher the antibacterial effect.

Treatment concentration part Minimum inhibitory concentration (MIC, g / ml) S. aureus P. aeruginosa E. Coli C. albicans A. niger One % Annona squamosa Taurus 315 276 122 271 39 Flower 350 289 132 273 40 leaf 325 302 125 277 42 stem 370 310 130 276 46 Root 370 307 133 280 43  Eupomatia laurina Taurus 245 211 98 231 31 Flower 252 230 100 233 35 leaf 266 225 106 235 33 stem 272 221 101 250 34 Root 280 238 103 251 39 magnolia Taurus 363 297 154 298 49 Flower 366 305 157 300 52 leaf 383 310 160 305 56 stem 400 313 163 303 57 Root 403 316 169 301 59 Magnolia tree Taurus 281 231 132 255 38 Flower 286 232 133 259 41 leaf 290 235 135 261 39 stem 310 238 140 263 39 Root 315 233 138 259 37  Tulip tree Taurus 497 439 298 351 198 Flower 508 440 299 353 199 leaf 515 441 297 354 199 stem 520 443 297 352 201 Root 525 439 296 353 198   Shark tree Taurus 377 298 169 401 121 Flower 380 299 169 418 120 leaf 400 300 172 405 121 stem 460 301 172 405 121 Root 425 298 171 406 120  Cherry tree Taurus 534 513 331 399 273 Flower 550 515 331 400 273 leaf 558 513 333 405 275 stem 500 512 335 399 279 Root 525 515 333 401 278  Beech tree Taurus 433 319 197 436 151 Flower 463 319 197 436 151 leaf 460 330 190 433 153 stem 450 320 190 436 152 Root 453 321 193 450 155

As shown in Table 18, all of the cell culture extracts of 1% Magnoliae representative plant-derived parts exhibited antibacterial effects, and particularly, the cell culture extracts from the taeylma showed excellent antimicrobial activity.

In addition, 200 μM methyl jasmonic acid inducer treatment and high frequency cell culture were performed for 3 days at 240-270 kHz for 5 days at a frequency of 240-270 kHz, In the case of the cells, the extract of the tubercle cell culture showed 1.5 times more antimicrobial effect than that of the untreated case, and the other cell culture extract also showed an increase of 1.1 times.

Experimental Example  11: Acne improvement effect

Ten men and women who developed acne were asked to apply the lotion thoroughly over the face for 4 weeks in the morning and evening. The results of the evaluation of the acne healing effect were evaluated according to the degree of the person who participated in the experiment by evaluating the pre-test condition by 1-3 points with reference to the following criteria, and the results are shown in the table.

 ≪ Evaluation of state before test >

 1 = a little acne 2 = a little acne 3 = acne severe

 ≪ Determination of acne effect &

-: Almost no effect, +: slight effect, ++: Much mitigated, +++: Completely healed

Treatment concentration part Pre-test condition Acne effect determination Effect after 2 weeks Effect after 4 weeks 2 % Annona squamosa Taurus Subject 1 2 + + Subject 2 One + ++ Subject 3 One - + Subject 4 2 - - Subject 5 2 + ++ Subject 6 2 + + Subject 7 3 ++ +++ Subject 8 2 - ++ Subject 9 3 + + Subject 10 2 + ++   Eupomatia laurina Taurus Subject 1 2 + ++ Subject 2 One + + Subject 3 One ++ +++ Subject 4 2 - ++ Subject 5 2 + + Subject 6 2 + ++ Subject 7 3 + + Subject 8 2 + ++ Subject 9 3 - + Subject 10 2 - -  magnolia Taurus Subject 1 2 + ++ Subject 2 One + + Subject 3 One + ++ Subject 4 2 + ++ Subject 5 2 - + Subject 6 2 - - Subject 7 3 + ++ Subject 8 2 + + Subject 9 3 + ++ Subject 10 2 + ++ Magnolia tree Taurus Subject 1 2 + ++ Subject 2 One + + Subject 3 One ++ +++ Subject 4 2 - ++ Subject 5 2 + + Subject 6 2 + ++ Subject 7 3 + + Subject 8 2 + ++ Subject 9 3 - + Subject 10 2 - - Tulip tree Taurus Subject 1 2 + + Subject 2 One + ++ Subject 3 One - + Subject 4 2 - - Subject 5 2 + ++ Subject 6 2 + + Subject 7 3 ++ +++ Subject 8 2 - ++ Subject 9 3 + + Subject 10 2 + ++ Shark tree Taurus Subject 1 2 + ++ Subject 2 One + + Subject 3 One ++ +++ Subject 4 2 - ++ Subject 5 2 + + Subject 6 2 + ++ Subject 7 3 + + Subject 8 2 + ++ Subject 9 3 - + Subject 10 2 - -  Cherry tree Taurus Subject 1 2 + + Subject 2 One + ++ Subject 3 One + + Subject 4 2 + ++ Subject 5 2 - + Subject 6 2 - - Subject 7 3 + + Subject 8 2 + ++ Subject 9 3 + + Subject 10 2 + ++ Beech tree Taurus Subject 1 2 + + Subject 2 One + ++ Subject 3 One - + Subject 4 2 - - Subject 5 2 + ++ Subject 6 2 + + Subject 7 3 ++ +++ Subject 8 2 - ++ Subject 9 3 + + Subject 10 2 + ++

As can be seen from Table 19 above, the improvement effect of acne was confirmed in the majority of subjects after 4 weeks of use of the softened lotion containing 2% magnolia thunb. Cell culture extract.

In addition, 200 μM methyl jasmonic acid inducer treatment and high frequency cell culture were performed for 3 days at 240-270 kHz for 5 days at a frequency of 240-270 kHz, In the case of the cells, the extract of the tubercle cell culture showed an antimicrobial effect twice as good as that of the untreated case and the effect of the other cell culture extract was improved by 1.5 times.

Preparation of cosmetic composition

Cosmetics of emulsified form such as nutritional lotion, cream, essence, etc. and cosmetics of solubilized form such as softened longevity were prepared with the cosmetic product containing the Magnoliaceae cell culture extract of Magnolia sieboldii of the present invention as an active ingredient.

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 Magnolia thyroid cell 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 Magnolia thyroid cell 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 Magnolia thyroid cell 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 Magnolia thyroid cell 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 Magnolia thyroid cell 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 (9)

A composition for external application for skin, which comprises an extract of a Liriodendron tulipifera teat cell culture, or a Tulip tree (Liriodendron tulipifera) teat cell culture.
The composition of claim 1, wherein the composition is anti-wrinkle or anti-wrinkle.
The composition of claim 1, wherein the composition is anti-inflammatory.
The composition of claim 1, wherein the composition is for shrinking pores.
The composition of claim 1, wherein the composition is for inhibiting sebum secretion.
The composition of claim 1, wherein the composition is for improving acne.
The composition of claim 1, wherein the composition is for skin moisturization.
The composition of claim 1, wherein the composition is a whitening agent.
2. The composition of claim 1, wherein the composition is antibacterial.