KR20110032670A - Composition for improving skin containing callus of broussonetia plant - Google Patents

Abstract

PURPOSE: A composition containing callus of Broussonetia kazinoki plants is provided to promote skin regeneration, moisturizing, and whitening and to prevent wrinkle and aging. CONSTITUTION: A composition contains 0.0001-25.0 weight% of crushed callus of Broussonetia kazinoki, callus culture liquid, callus extract, crushed material of cells differentiated from the callus, culture liquid of the cells, or extract of the cells as an active ingredient. The composition is used in skin regeneration, anti-wrinkling, skin whitening, moisturizing, or skin tone improvement.

Description

Composition for improving skin containing callus of Broussonetia plant}

The present invention relates to a composition containing the callus of the genus Hakdan.

Plants used to mean food resources in the past, but now they are expanding as a source of a wide range of chemicals, including pharmaceuticals, flavorings, pigments, pesticides, and dyes. In particular, most of the plant-derived useful substances have physiological activities such as antiviral, antibacterial, anticancer, and anti-oxidant abilities, attracting attention as an ideal resource that can be developed as a novel efficacious substance. It also has the advantage of being friendly.

However, there are practical difficulties in commercial development of novel potent substances from plant sources. The reason for this is as follows. Plants grow very slowly, the content of potent substances in plants is extremely limited, potent substances in plants are present only in certain organs of the plant, and problems with natural degradation occur when plants are used as resources. And finally, plant-derived agonists have a very complicated chemical structure, requiring multi-stage polymerization processes, resulting in economic problems that lead to very high production costs. For this reason, it was difficult to stably supply potent substances derived from plants.

The present invention promotes skin regeneration of at least one selected from the group consisting of callus lysate, callus culture medium, callus extract, lysate of cells differentiated from callus, culture solution of cells differentiated from callus and extracts of cells differentiated from callus , Anti-wrinkle, skin whitening, skin moisturizing, skin tone improvement, anti-aging or improvement of skin wrinkles, callus shreds of callus, callus culture, callus extract, shreds of cells differentiated from callus, A skin condition improving composition containing as an active ingredient at least one selected from the group consisting of cultures of cells differentiated from callus and extracts of cells differentiated from callus.

The present invention comprises at least one selected from the group consisting of callus lysate, callus culture medium, callus extract, lysate of cells differentiated from callus, culture medium of cells differentiated from callus and extract of cells differentiated from callus as active ingredients The composition containing is disclosed.

In one embodiment of the present invention, the composition is characterized in that it has a purpose of promoting skin regeneration.

In one embodiment of the present invention, the composition is characterized in that it has a purpose of preventing or improving skin wrinkles.

In one embodiment of the present invention, the composition is characterized in that it has a skin whitening use.

In one embodiment of the invention, the composition is characterized in that it has a skin moisturizing use.

In one embodiment of the invention, the composition is characterized in that it has a skin tone improvement use.

In one embodiment of the present invention, the composition is characterized in that it has an anti-aging or improving use.

The present invention comprises at least one selected from the group consisting of callus lysate, callus culture medium, callus extract, lysate of cells differentiated from callus, culture medium of cells differentiated from callus and extract of cells differentiated from callus as active ingredients It has the effect of promoting skin regeneration, preventing or improving skin wrinkles, skin whitening, skin moisturizing, skin tone improvement, anti aging or improvement.

As used herein, the term "skin" refers to a tissue covering the body surface of an animal, and is a broad concept including not only tissues covering the body surface such as the face or body, but also the scalp and hair.

Hereinafter, the present invention will be described in detail.

With the development of biotechnological techniques, the field of induced differentiation of undifferentiated cells has recently emerged, and many experiments related to induced differentiation of undifferentiated cells have been conducted, such as the study of signals involved in the development process. The plant cell culture method using the same has emerged as an ideal technology for stably supplying useful materials derived from plants without causing environmental problems. The production of useful substances by plant cell culture has numerous advantages over methods by direct extraction from plants. In particular, unlike conventional extraction methods, it is possible to continuously produce without being influenced by the external environment, and thus it is considered to be the best way to solve the problems such as ecosystem destruction.

When harvesting leaves or stems to obtain potent substances from plants, harvesting can be continued without damaging the plant, but when roots are harvested, the whole plant is damaged and the plant can no longer survive. . However, when using differentiated cells, even a small amount of plant cells can easily obtain a large amount of cells differentiated into root tissues, so that raw materials of agonists can be obtained without damaging the plant, thereby protecting the ecosystem while protecting the ecosystem. It is possible to establish a green process for the raw material.

Mulberry is a mulberry family Broussonetia kaempferi , Broussonetia kazinoki ( doughwood ), Broussonetia kurzii , Broussonetia Contains papyrifera (ketchup). In the present invention, the mulberry plant includes the four species, more specifically in the mulberry ( Broussonetia kazinoki ) and beech ( Broussonetia papyrifera ). These are deciduous broad-leaved shrubs in the mulberry tree, and are distributed in most parts of Korea (mostly in the south), China, Taiwan, and Japan. It has long been known that bask fiber is used as a raw material of paper, and it has been known to have various effects. It is reported to be effective in tonic, nominal function, genital, species, ripening and diuresis, remove wind, clear blood and reduce vision.

The present invention comprises at least one selected from the group consisting of callus lysate, callus culture medium, callus extract, lysate of cells differentiated from callus, culture medium of cells differentiated from callus and extract of cells differentiated from callus as active ingredients The composition containing is disclosed. The composition has the effect of promoting skin regeneration, preventing or improving skin wrinkles, skin whitening, skin moisturizing, skin tone improvement, anti-aging or improvement. Kazinol is present in the cells differentiated from the callus or callus of the mulberry plant. This has the advantage of obtaining a raw material of a potent substance without damaging the plant by using the cells differentiated from the callus or callus of the plant of the mulberry plant itself, not the mulberry itself. It can also have better efficacy than the mulberry plant itself.

As used herein, "callus" refers to a cell mass of an early stage cultured in solid medium by inducing callus. "Cells differentiated from cells" refers to the early stages of callus-induced incubation in solid medium, to the process of treating plant hormones to differentiate them into specific organs. In one embodiment of the invention the specific organ may be a leaf, stem or root. In an embodiment of the present invention, cells differentiated from callus include cells differentiated from callus to leaf, cells differentiated from callus to stem, and cells differentiated from callus to root. Typically, the time required for differentiation to specific organs of woody plants is about 1 day to 60 days, specifically, may have a differentiation period between 1 day and 45 days.

As used herein, the term “crushed product” refers to a cell lysate obtained by crushing a cell by a chemical method using a general detergent or the like, or a physical method using a French press, and the like. It may be prepared according to a known conventional method, and the method is not particularly limited.

As used herein, the term "culture medium" refers to the remaining culture solution from which pellets are removed using a physical method such as ultra-centrifugation after undergoing a process of culturing the cells for 6 hours to 72 hours by a general method in the art. The method may be prepared according to conventional methods known in the art, and the method is not particularly limited.

As used herein, the term “extract” refers to a substance obtained by dissolving cells in a solvent and concentrated by distillation or evaporation. The method may be prepared according to a conventional method known in the art, and the method It is not specifically limited. Generally, after drying and pulverizing, an extraction solvent is added and heated to extract the active ingredient and concentrated under reduced pressure to obtain an extract. As the extraction solvent, a solvent that can be generally used may be used, and may include water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate, butyl acetate, 1,3-butylene glycol, and the like.

The cell culture method used in the present invention is a known solid culture method. Solid culture is a culture of callus cells on a solid medium. Callus can be induced as follows: Cut leaves and stems surface-treated with sodium hypochlorite and ethyl alcohol to an appropriate size and cultivate aseptically on a solid culture medium. do. After a few weeks of culture, callus cells are formed at the cutting surface of the roots and leaves and begin to grow. The callus is passaged continuously to establish a stable cell line.

The medium used for inducing callus cells may be MS (Murashige & Skoogs) medium. 0.5-1.0% of agar is added to this MS medium to use as a solid medium. The plant hormone auxin is added as a growth regulator. As auxin, 1-naphthaleneacetic acid (NAA) can be used instead of 2,4-D, which is seriously harmful to humans. Cell growth is also good when the sucrose concentration is maintained at 30 g / 1 or less in the medium.

The culture temperature of 20-25 ℃ is suitable, the growth of cells under dark conditions is better than the light conditions with light. Under these conditions, callus cells growing in solid medium are passaged to fresh medium every 4 to 5 weeks, during which time the amount of cells increases by about 2 to 3 times.

Cells differentiated from the callus of the present invention can be prepared according to a conventional method known in the art as cells differentiated from roots, stems, and leaves in the callus, wherein auxin, gibberellic acid, phyto as a growth regulator in the medium Hormones and the like may be added in different concentrations as appropriate for the organs to differentiate to a concentration between 0.1 ~ 3mg / L.

The skin is divided into three layers of epidermis, dermis, and subcutaneous fat tissue in order from the outside, and the body's primary protective film protects the organs from changes in temperature and humidity, irritation from the external environment such as ultraviolet rays and pollutants. It plays an important role in maintaining homeostasis such as regulation. However, ultraviolet rays, excessive physical and chemical stimuli from the outside, stress, and nutritional deficiency decrease the normal function of the skin and promote skin aging such as loss of elasticity, keratinization and wrinkle formation. In particular, if keratinocytes are made in sequence and the outermost layers of old keratinocytes sometimes break away, the keratinization process, which causes the epidermis to maintain a constant thickness, is not successful, creating abnormal keratinocytes and thickening of the stratum corneum. The skin becomes rough and dull, fine wrinkles are generated and eventually aging of the skin. Therefore, when promoting the growth of keratinocytes, not only can promote skin regeneration, but also prevent and improve wrinkles, have a skin whitening effect, improve skin tone, and further prevent or improve aging.

Collagen refers to an important fibrous protein of animal connective tissue, which has the function of preventing or improving skin wrinkles, skin regeneration, preventing or improving scars, strengthening skin elasticity, and preventing skin aging. Procollagen is a precursor of collagen found in the skin and other organs of many vertebrates, resulting in the same function as collagen. Thus, elevated levels of collagen and procollagen may prevent or improve skin wrinkles, regenerate skin, prevent or improve scars, enhance skin elasticity, and prevent or improve skin aging. Meanwhile, "collagenase" is an enzyme that breaks down collagen and is known as one of the causes of skin wrinkles. Collagenase (Matrix metalloproteinase-1, MMP-1) can be biosynthesized by ultraviolet light associated with the degradation of the skin matrix, which leads to a decrease in the biosynthesis of type 1 procollagen. Therefore, inhibiting the expression of MMP-1 increases the collagen level of the skin, which can also prevent or improve skin wrinkles, regenerate the skin, strengthen skin elasticity, and prevent or improve skin aging. In addition, the diminished capacity of fibroblasts present in the dermal layer of skin is also associated with skin aging.

Melanin is a pigment that determines the color of skin and hair, and melanin produced from melanocytes moves out of the skin and is distributed in the stratum corneum. Exposure to UV rays increases the number of melanocytes and increases the production of melanin, causing pigmentation, blemishes, and freckles. Diseases such as Becker's nevus, lentigines and Nevus spilus are also caused by excessive deposition of epidermal melanin.

Melanin is a complex produced by complex enzymatic and non-enzymatic oxidation of tyrosine, and it is known that tyrosinase plays an important role in this process. Tyrosinase is a metal protein enzyme that binds to copper. It is widely distributed in animals, plants, microorganisms, and humans. Tyrosine is dihydroxy phenylalanine (dopa, dihydroxyl-L-phenylalanine, L-DOPA), and dihydroxy phenylalanine. It acts to convert dopaquinone into dopaquinone. Dopaquinone is converted to dopachrome, and dopachrome is converted to dihydroxyindole and finally synthesized melanin through oxidative polymerization of dihydroxyindole. Therefore, inhibition of tyrosinase activity or inhibition of L-DOPA can inhibit melanin synthesis. Inhibiting melanin production has a skin whitening effect, improves skin tone, and prevents or prevents aging.

Most widely used whitening agents contain tyrosinase inhibitors or tyrosinase inhibitors, such as arbutin, hydroquinone, hydroxyanisole, ascorbic acid derivatives, and kojic acid. ), Corticosteroids, retinoids, etc., but some inhibitors have been reported to be carcinogenic by causing mutations. In addition, there are safety, economical, and transdermal absorption problems. There is difficulty. In particular, hydroxyanisole, hydroquinone and the like have strong melanogenesis inhibitory activity, but have side effects such as causing denaturation or lethality of pigment cells and impairing the intrinsic function of cells. In addition, kojic acid has problems such as low inhibitory activity, discoloration during use, and instability of the substance itself.

The present invention comprises at least one selected from the group consisting of callus lysate, callus culture medium, callus extract, lysate of cells differentiated from callus, culture solution of cells differentiated from callus and extracts of cells differentiated from callus as an active ingredient To promote keratinocyte growth, inhibit MMP-1, increase procollagen, promote tyrosinase activity, inhibit L-DOPA oxidation, inhibit melanin production, promote skin regeneration, prevent skin wrinkles or The present invention relates to a composition having an improvement, skin whitening, skin moisturizing, skin tone improvement, anti-aging or improvement effect.

At least one selected from the group consisting of callus lysate, callus culture medium, callus extract, lysate of cells differentiated from callus, culture solution of cells differentiated from callus and extracts of cells differentiated from callus of the birch plant of the present invention It may be contained in an amount of 0.0001 to 25.0% by weight. If the content is less than 10,000% by weight, the effect is insignificant, and if the content is more than 25.0% by weight may cause problems in the feel of the product and the stability of the formulation.

The present invention comprises at least one selected from the group consisting of callus lysate, callus culture medium, callus extract, lysate of cells differentiated from callus, culture medium of cells differentiated from callus and extract of cells differentiated from callus as active ingredients The cosmetic composition containing the composition containing can be provided. Cosmetic compositions contain a cosmetically or dermatologically acceptable medium or base. These are all formulations suitable for topical application, e.g. emulsions obtained by dispersing the oil phase in solutions, gels, solids, pasty anhydrous products, aqueous phases, emulsions obtained by dispersing the aqueous phase in oil phases, multiemulsions, suspensions, micro emulsions, micro It may be provided in the form of capsules, microgranules, or ionic (liposomal) and nonionic vesicle dispersants, or in the form of creams, skins, lotions, powders, ointments, sprays, cleansers or concealed sticks. These compositions may be prepared according to conventional methods in the art. The composition according to the invention can also be used in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

The cosmetic composition may contain, in addition to the above-mentioned substances, other ingredients that can give a synergistic effect to the main effect, preferably within the range that does not impair the main effect of the present invention, in addition to the active ingredient of the present invention. Other ingredients can be suitably selected and combined with those skilled in the art according to the formulation or use purpose of the other cosmetic composition.

For example, the cosmetic composition may contain a skin absorption promoting substance to increase the effect. In addition, the cosmetic composition of the present invention may include a material selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids and seaweed extract. In addition, the cosmetic composition of the present invention may include other components which are usually formulated into the cosmetic composition as necessary, in addition to the essential components, and examples thereof include fats and oils, humectants, emollients, surfactants, organic and inorganic. Pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusting agents, alcohols, pigments, flavoring agents, blood circulation accelerators, cooling agents, limiting agents, purified water and the like. Other compounding components that can be included in the cosmetic composition of the present invention is not limited thereto, and the compounding amount of the above components is possible within a range that does not impair the object and effect of the present invention.

The cosmetic composition is not particularly limited in formulation, and may be appropriately selected in accordance with the desired purpose. For example, flexible lotion (skin lotion and milk lotion), nourishing lotion, essence, nourishing cream, massage cream, powder, lipstick, makeup base, foundation, patch, spray, pack, gel, essence, eye cream, eye essence, It may be prepared in one or more formulations selected from the group consisting of cleansing cream, cleansing foam, cleansing water, cleanser, body lotion, body cream, body oil and body essence, but is not limited thereto.

The present invention comprises at least one selected from the group consisting of callus lysate, callus culture medium, callus extract, lysate of cells differentiated from callus, culture medium of cells differentiated from callus and extract of cells differentiated from callus as active ingredients A pharmaceutical composition can be provided comprising the composition containing it.

When the composition according to the present invention is applied to a medicament, the composition can be formulated into an oral or parenteral dosage form in the form of solid, semi-solid or liquid by adding a commercially available inorganic or organic carrier. If the active ingredient of the present invention is carried out according to the conventional method, it can be easily formulated, and it can be easily formulated with surfactants, excipients, colorants, spices, stabilizers, preservatives, preservatives, wetting agents, emulsifiers, suspending agents, salts for controlling osmotic pressure and / or Buffers and other commercial auxiliaries can be used as appropriate.

As the preparation for oral administration, tablets, pills, granules, soft and hard capsules, powders, fine granules, powders, solutions, emulsions, syrups, pellets, etc. Can be mentioned. These formulations contain, in addition to the active ingredient, diluents (e.g. lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and glycine), glidants (e.g. silica, talc, stearic acid and its magnesium or calcium salts and polyethylene glycols) It may contain. Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidine, optionally starch, agar, alginic acid or its sodium salt Pharmaceutical additives such as disintegrants, absorbents, colorants, flavors, and sweeteners. The tablets can be prepared by conventional mixing, granulating or coating methods.

In addition, the parenteral administration agent may be, for example, an external preparation for skin, and may be an injection, drop, lotion, ointment, gel, cream, suspension, emulsion, suppository, patch or spray formulation. It is not limited to this.

The pharmaceutical composition according to the present invention may be administered orally, parenteral, rectal, topical, transdermal, intravenous, intramuscular, intraperitoneal, subcutaneous.

In addition, the dosage of the active ingredient will vary depending on the age, sex and weight of the subject to be treated, the specific disease or pathology to be treated, the severity of the disease or pathology, the route of administration and the judgment of the prescriber. Dosage determination based on these factors is within the level of skill in the art. Typical dosages are 0.001 mg / kg / day to 2000 mg / kg / day, more specifically 0.5 mg / kg / day to 2.5 mg / kg / day.

Hereinafter, the structure and effects of the present invention will be described in more detail with reference to Preparation Examples, Experimental Examples, Examples and Comparative Examples. However, these Preparation Examples, Experimental Examples, Examples and Comparative Examples are provided only for the purpose of illustration in order to facilitate understanding of the present invention, the scope and scope of the present invention in the following Preparation Examples, Experimental Examples, Examples and Comparative Examples It is not limited by.

Preparation Example Callus Culture and Preparation of Callus Fragment, Culture Solution, Extract

Roots and leaves with good growth status among the collected mulberry and cotylidum were used as in vivo samples for callus induction. The biological sample was cut to about 10 cm in length, washed, and then placed in a 0.5% sodium hypochlorite solution containing 0.1% of Tween-20, followed by surface sterilization for 20 minutes with stirring. During this period, ultrasonic sonication was performed for 3 seconds to remove microbubbles on the surface of the biological sample. The plate was washed once with sterile water and then sterilized again in a 70% ethanol solution for 1 minute, and then the sterilized and washed biological samples were cut to a size of about 0.5 cm, and 0.5-1.0% of agar was added to MS medium. The culture was started by transferring the mixed solid culture medium. As a growth regulator, 1-naphthalene acetic acid (NAA), a kind of plant hormone auxin, was used. In addition, the concentration of sucrose (sucrose) in the medium was maintained below 30g / l.

All manipulations were performed aseptically on a sterile operating table, and callus was induced from 2-3 weeks after incubation at 25 ° C dark conditions. Callus was well derived from various biological samples, and the excellent one was transferred to fresh medium every month to establish a cell line.

Callus extract is centrifuged to clean the cell line, washed with sterile purified water, sterile dried cells (weight 100g) was pulverized, to the pulverized product was added 0.5 L of 95% ethanol, the extractor attached to the cooling capacitor It was prepared by extracting by heating at 45 ℃ for 3 days at. The extract was filtered through a 350 mesh filter cloth and then filtered through Whatman No. 2 filter paper to use in the following experiment.

On the basis of the established cell lines, callus lysates and callus cultures of mulberry and coriander were prepared in a general manner.

 Experimental Example 1 Evaluation of keratinocyte growth promoting effect (MTT assay)

Effects of Callus Crushes, Callus Cultures, Callus Extracts, Callus Extracts from Cellulose from Cellulose and Cellulose, Cultures, Extracts, Calligraphy from Cells from Cells, Cultures, and Extracts on Growth Capacity of Keratinocytes Was measured by MTT method.

First, the keratinocytes were planted in a 96 well plate at a concentration of 5 × 10 ³ per 200 μl per well and incubated for 24 hours, followed by callus lysate, callus culture medium, callus extract, and callus Cultivates, cultures, extracts of cells differentiated into the roots, cells, cells, and extracts of cells differentiated from stems to callus were incubated for 48 hours by treating with 0.001, 0.01 and 0.1%, respectively, and then inhaled and removed the cultures. Washed once with PBS, MTT (Methylthiazolyldiphenyl-tetrazolium bromide, Sigma, USA) (0.5 mg / mL) solution was added to the cells 100 μl and incubated at 37 ° C., 5% CO ₂ for 4 hours. Thereafter, the culture solution was aspirated and removed, and then 200 µl of DMSO (dimethyl sulfoxide) solution was added, shaken for 10 minutes in a shaker, and the absorbance at 540 nm was read with an ELISA reader (DIbiotech, Korea). . At this time, the control group was used as a non-treated group treated nothing, the results are shown in Table 1 below. Higher absorbance indicates an effect of promoting growth of keratinocytes.

density(%) Absorbance Paper mulberry Coriander Callus Shreds 0.001% 135 138 0.01% 136 141 0.1% 143 154 Callus culture 0.001% 148 135 0.01% 152 137 0.1% 162 140 Callus extract 0.001% 113 115 0.01% 115 119 0.1% 118 124 Crushes of cells differentiated from callus to root 0.001% 125 128 0.01% 126 131 0.1% 138 137 Culture of cells differentiated from callus to root 0.001% 122 121 0.01% 125 125 0.1% 129 126 Extract of Cells Differentiated from Callus to Roots 0.001% 112 114 0.01% 115 118 0.1% 121 123 Remnants of cells differentiated from callus to stem 0.001% 115 121 0.01% 121 127 0.1% 127 129 Culture of cells differentiated from callus to stem 0.001% 128 128 0.01% 130 132 0.1% 138 139 Extract of Cells Differentiated from Callus to Stem 0.001% 103 106 0.01% 103 108 0.1% 106 110 Untreated control 0% 100 101

As shown in Table 1, callus lysate, callus culture medium, callus extract, callus lysate of the mulberry and koji tree used as an active ingredient in the present invention, lysate of cells differentiated from roots in callus, culture solution, extract, callus differentiated into stems Cell debris, culture medium, extracts were all excellent in promoting the growth of keratinocytes. In addition, as the concentration of each component increased, the effect of promoting the division of keratinocytes also increased.

Callus lysate, callus culture medium, callus extract, lysate of cells differentiated from callus to root, cultures, extracts, callus lysate from stems of callus , Cultures, extracts have the effect of promoting the regeneration of the skin, prevent and improve skin wrinkles, improve skin tone, and further prevent and improve aging. These can be used in mixture of 2 or more types.

 Experimental Example 2 Evaluation of MMP-1 Inhibition and Procollagen Promoting Effect (procollagen assay)

To investigate the effects of callus lysate of callus and cotyledon, callus culture, lysate of cells differentiated from root to callus, cultures of cells differentiated from root to callus, and MMP-1 biosynthesis and promoting the biosynthesis of type 1 procollagen MMP-1 and procollagen assays were performed in the following manner.

First, the fat layer of the normal epidermis is removed and chopped to separate the epidermis and the dermis with collagenase, and then the epidermis and the dermal tissue are placed in 0.25% trypsin solution, respectively, at 37 ° C. and 5% CO ₂ incubator. Treated for a minute. Since vortex was performed to release the keratinocytes and fibroblasts, respectively. After collecting and washing the free cells, keratinocytes were cultured at 1 × 10 ⁴ cells / cm ² in KGM (keratinocyte growth medium, Clonetics, USA), and fibroblasts were added with 10% fetal bovine serum (FBS). Cultured in DMEM medium. When grown at about 70-80%, the cells were subcultured at a ratio of 1: 3, and cells subjected to 3-4 subcultures were used for the experiment.

For experiments to measure the amount of MMP-1 fibroblasts were cultured in more than 90% in a 48 well plate (star plate) was placed in a starvation (starvation) state for one day. After washing twice with PBS, the lid of the plate (plate) was opened with 100 μl of PBS and irradiated with UV A (Dermlight cube 401, UVAtec, USA) equipped with UVA filters at 15 J / cm ² . Once again immediately after irradiation, washed once with PBS, callus lysate of 0.001%, 0.01%, and 0.1% of mulberry and coriander, callus culture, lysate of cells differentiated from callus to root, and culture of cells differentiated from callus to root, respectively. This fetal calf serum was added to DMEM without the same, and cultured for 48 hours in a 37 ° C., 5% CO ₂ incubator, and then the amount of free MMP-1 in the medium was determined using the MMP-1 human ELISA system. system) (RPN2610, Amersham Pharmacia Biotech, UK) and calibrated with the total protein amount of fibroblasts. At this time, a non-treated group that was not treated with anything was used as a control group, the results are shown in Table 2.

Meanwhile, in order to measure the amount of procollagen, callus crushed matters of 0.001%, 0.01%, and 0.1% of mulberry and coriander without UV irradiation in starvation conditions under the same conditions as the method of measuring the amount of MMP-1, respectively. , Callus culture, lysate of cells differentiated from roots to callus, cultures of cells differentiated from roots to callus were added, and after 24 hours, the amount of free collagen released in the medium was determined using the procollagen type 1 C-peptide EIA kit (procollagen). Measurement was performed using a type-1 C-peptide EIA kit (MK101, Takara, Japan). At this time, a non-treated group that was not treated with anything was used as a control group, the results are shown in Table 3.

density(%) Amount of MMP-1 Paper mulberry Coriander Callus Shreds 0.001% 70 75 0.01% 66 68 0.1% 62 60 Callus culture 0.001% 72 75 0.01% 71 71 0.1% 66 64 Crushes of cells differentiated from callus to root 0.001% 65 74 0.01% 61 61 0.1% 54 53 Culture of cells differentiated from callus to root 0.001% 67 70 0.01% 63 64 0.1% 61 60 Untreated control 0% 100 100

density(%) Amount of procollagen Paper mulberry Coriander Callus Shreds 0.001% 117 115 0.01% 125 125 0.1% 129 134 Callus culture 0.001% 107 117 0.01% 115 124 0.1% 127 129 Crushes of cells differentiated from callus to root 0.001% 126 127 0.01% 130 132 0.1% 137 137 Culture of cells differentiated from callus to root 0.001% 125 123 0.01% 129 130 0.1% 135 136 Untreated control 0% 100 100

As shown in the results of Table 2, callus shreds of callus and koji tree used as an active ingredient in the present invention, callus culture medium, shreds of cells differentiated from callus to root, cultures of cells differentiated from callus to root It was found that it inhibits MMP-1. Also, as the concentration of each component increased, the amount of MMP-1 decreased proportionally.

As shown in the results of Table 3, callus shreds of callus and coriander used as active ingredients in the present invention, callus culture medium, shreds of cells differentiated from callus to root, cultures of cells differentiated from root to callus are all It was found to increase the biosynthesis of type 1 procollagen. In addition, as the concentration of each component increased, the amount of procollagen produced increased proportionally.

Callus lysate of callus and cotyledon, callus culture medium, lysate of cells differentiated from callus to root, and cultures of cells differentiated from callus to root have excellent MMP-1 inhibitory effect and procollagen biosynthesis promoting effect. Promotes and prevents or improves wrinkles, and further prevents or improves aging. These can be mixed and used 2 or more types.

Experimental Example 3 Evaluation of Tyrosinase Activity Inhibition Activity

Enzyme tyrosinase was extracted from mushrooms, and SIGMA Co., Ltd. was used. First, the substrate tyrosine was dissolved in distilled water to make a 0.3 mg / ml solution, and the solution was added to the test tube by 1.0 ml, and 1.0 ml of potassium-phosphate buffer solution (0.1 mol, pH 6.8) and 0.7 ml of distilled water were added thereto. It was. 0.2 ml of callus lysates, cultures, extracts of mulberry and coriander, lysates of cells differentiated into roots from callus of mulberry and coriander, lysates of cells differentiated into leaves from callus of mulberry, from callus of stems from coriander The lysate, the mulberry extract, and the locust extract of the cells were added to the reaction solution and reacted for 10 minutes in a 37 ° C thermostat. In this case, only 0.2 ml of the solvent was added to the control instead of the test substance. 0.1ml each of 2500Unit / ml tyrosinase solution was added to the reaction solution and reacted for 10 minutes in a 37 ° C thermostat. The test tube containing the reaction solution was placed in iced water and quenched to stop the reaction, and the absorbance at 475 nm was measured with a spectrophotometer. The positive control group used the existing whitening agents arbutin and ascorbic acid.

Tyrosinase inhibitory effect of each test substance was evaluated by calculating the tyrosinase inhibition rate by the following equation (1), the results are shown in Table 4.

% Tyrosinase inhibition rate = 100-{(reaction absorbance of each test substance / control absorbance of control) × 100}

Treatment concentration (%) Tyrosinase Inhibition Rate (%) Paper mulberry callus shreds 0.001% 23.1 0.01% 29.0 0.1% 34.6 Cucumber Callus Shreds 0.001% 23.2 0.01% 27.4 0.1% 36.1 Paper mulberry callus 0.001% 12.8 0.01% 16.4 0.1% 21.7 Cucumber callus culture 0.001% 14.3 0.01% 17.9 0.1% 21.1 Paper mulberry callus extract 0.001% 12.1 0.01% 14.7 0.1% 19.2 Coriander Callus Extract 0.001% 14.2 0.01% 16.3 0.1% 17.8 Crushes of Cells Differentiated from Callus of Paper mulberry to Roots 0.001% 75.3 0.01% 88.7 0.1% 96.3 Culture of Differentiated Cells from Roots of Callus of Paper mulberry 0.001% 15.6 0.01% 19.2 0.1% 31.9 Extract of Cells Differentiated from Roots of Callus of Paper mulberry 0.001% 14.2 0.01% 21.2 0.1% 26.9 Crushed Cells Differentiated from Roots of Callus to the Roots 0.001% 95.2 0.01% 96.3 0.1% 99.1 Remnants of Cells Differentiated to Leaves in Callus of Paper mulberry 0.001% 4.1 0.01% 8.7 0.1% 10.0 Crushed Cells Differentiated into Stems from Callus of Coconut Tree 0.001% 3.2 0.01% 6.5 0.1% 23.5 Paper mulberry extract 0.01% 33.1 Coriander Extract 0.01% 40.4 Ascorbic acid 0.01% 39.2 Arbutin 0.01% 50.4

As can be seen in Table 4, the callus shreds, culture medium, extracts of the mulberry and coriander used as active ingredients in the present invention, the shredded products of the cells differentiated into the roots from the callus of the mulberry and coriander, culture solution, extract, mulberry The inhibitory effect of tyrosinase activity of the lysates of the cells differentiated from the callus of the leaves into the leaves, and the lysates of the cells differentiated from the callus of the coriander to the stems was recognized. Among them, the inhibitory effect of tyrosinase activity of the lysates of the cells differentiated to the root from the callus of the mulberry and the locust tree was higher than the lysates of the cells differentiated from the callus of the mulberry to the leaves and of the cells of the callus of the locust tree. They showed a very good inhibitory effect on tyrosinase activity compared to the extracts of the mulberry and the cockroach. In addition, they had an effect of inhibiting tyrosinase activity superior to that of ascorbic acid and arbutin at the same concentration. And as the amount of each active ingredient increased, tyrosinase activity also tended to be suppressed.

Cells, cell cultures, extracts, lysates of cells differentiated to leaves from callus of mulberry, and cells differentiated to stems from callus of locust tree The crushed product of has a skin whitening effect, improves skin tone, and has an effect of preventing or improving skin aging. These can be mixed and used 2 or more types.

Experimental Example 4 Evaluation of the Antioxidant Activity of L-DOPA

The inhibitory effect of cell debris, mulberry extract, and cucurbita extract on the differentiation of the roots from the callus of mulberry and cotylodon on the L-DOPA (L-dopa) oxidation of tyrosinase catalyzed the rate-determination step of melanin synthesis Evaluated.

Disintegration of cell, branched and cotyledon extracts from the callus of mulberry and cotylidum was dissolved in DMSO and diluted again in 0.1M sodium phosphate buffer (pH 6.5) to give a final concentration of 16.67, 33.34, 66.67, 166.67, 333.33 and 666.67 μg / ml were prepared. 850 μl of 0.1 M phosphate buffer (pH 7.0), 50 μl of the above prepared solution, and 50 μl of mushroom tyrosinase (SIGMA) (1500 to 2,000) U / ml solution were placed in this order. L-DOPA (L-3) , 4-dihydroxyphenylalanine) solution was added and then reacted at 37 ° C. for 15 minutes. After the experiment, the absorbance was measured at 475 nm using a spectrophotometer. The inhibition rate of L-DOPA oxidation activity (%) was calculated as follows by Equation 2, and the results are shown in Table 5.

L-DOPA oxidative activity inhibition rate (%) = 100-{(reaction absorbance of each test substance / control absorbance of control) × 100}

Treatment concentration (%) L-DOPA oxidative activity inhibition rate (%) Crushes of Cells Differentiated into Roots of the Paper Mulberry Callus 0.001% 77.1 0.01% 80.4 0.1% 90.9 Crushed Cells Differentiated into Roots from Coriander Callus 0.001% 87.8 0.01% 93.3 0.1% 97.9 Paper mulberry root extract 0.01% 63.7 Coriander Root Extract 0.01% 75.8 Ascorbic acid 0.01% 51.1 Arbutin 0.01% 89.1

As can be seen from Table 5, it was recognized that the lysates of cells differentiated from the callus of the mulberry and coriander used as active ingredients in the present invention to the root have an excellent L-DOPA antioxidant inhibitory effect. They had a superior L-DOPA antioxidant activity than the root extracts of the mulberry and locust trees. In particular, the oxidation of L-DOPA was inhibited as the amount of lysate of the cells differentiated to the roots in callus of the mulberry and coriander tree was inhibited, and the addition of 0.1% inhibited the oxidation of L-dopa by almost 100%. The inhibitory effect of L-DOPA oxidation of the cell lysates differentiated from the callus of mulberry and koji was found to be superior to the arbutin widely used as a conventional whitening agent.

As described above, the debris of cells differentiated into roots in callus of mulberry and koji tree having L-dopa antioxidant effect has skin whitening effect, improves skin tone, and prevents or improves aging. These can be mixed and used 2 or more types.

Experimental Example 5 Evaluation of Melanin Inhibition

The inhibitory effect of melanin production on the cells differentiated from the roots of the callus of the mulberry and coriander was investigated. The untreated group treated with nothing and the control treated with only solvent DMSO were also tested.

After 5-6 ml of DMEM medium was placed in a 25 cm 2 T-flask, the mouse-derived B-16 melanoma (KCLB 80008) cell line was inoculated in a freezer and incubated for 24 hours at 37 ° C. and 5% CO ₂ . Cells were then separated by treatment with 0.05% trypsin containing 0.02% EDTA, inoculated in 6-well plates at 1 × 10 ⁵ per well and incubated until at least 80% of the cells adhered to the bottom of the well. After incubation, the medium was removed, and the cells of the callus of mulberry and coriander dissolved in DMSO were replaced with medium diluted to the final concentrations of 0.5, 5, and 50 µg / ml of the cell debris, mulberry extract, and coriander extract. 3 days were cultured in the% CO _2, 37 ℃. After 3 days, the cells from which the medium was removed were washed with PBS (phosphated buffer saline), treated with trypsin to recover the cells, centrifuged at 5,000 rpm for 10 minutes, and the supernatant was removed to obtain pellets. The cell pellet was dried at 60 ° C. for 2 hours, dissolved in 100 μl of 1 M sodium hydroxide solution containing 10% DMSO, and then absorbed at 490 nm using a spectrophotometer to determine the amount of melanin per cell protein. Synthetic melanin (Sigma, USA) was dissolved in 100 μl of 1 M sodium hydroxide solution containing 10% DMSO at various concentrations to obtain a standard concentration curve therefrom to determine the concentration of melanin. Based on this value, the percentage of melanin inhibition of cell debris, mulberry extract, and cucurbita extract from the callus of mulberry and beech was determined based on the untreated group. Among them, the results of the addition of the pulverized cells, the mulberry extract, and the beech extract of the cells differentiated into the roots of callus of the mulberry and the beech tree were added to the mouse-derived B-16 melanoma culture such that the final concentration was 50 µg / ml. It was.

As can be seen in Figure 1, the amount of melanin produced when added to the mouse-derived B-16 melanoma culture solution to the final concentration of 50 ㎍ / ㎖ of the cell differentiation to the root in the callus of the mulberry and coriander Significantly lower than the non-added control group. It was confirmed that the melanin production inhibition rate was lower due to less melanin amount of cell debris differentiated from the callus of the mulberry and the beech tree to the root than in the case of the arbutin, the mulberry extract, and the beech tree extract, which are widely used as the existing whitening agents. Therefore, it can be seen that the lysate of the cells differentiated into the roots of callus of the mulberry and coriander acts as an excellent whitening agent.

As described above, the debris of cells differentiated from roots of callus of the mulberry and koji tree having melanin production inhibitory effect has skin whitening effect, improves skin tone, and prevents or improves aging. These can be mixed and used 2 or more types.

 [Examples 1 to 8 and Comparative Example 1]

To the cosmetic composition of Examples 1 to 8 and Comparative Example 1 was prepared in the composition of Table 6.

Ingredients (content: wt%) Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Example 7 Example 8 Comparative Example 1 1.Beads wax 2 2 2 2 2 2 2 2 2 2.stearyl alcohol 3 3 3 3 3 3 3 3 3 3.stearic acid 8 8 8 8 8 8 8 8 8 4.squalene 10 10 10 10 10 10 10 10 10 5.propylene glycol monostearate 3 3 3 3 3 3 3 3 3 6.Polyoxyethylene ether One One One One One One One One One 7.preservative Quantity Quantity Quantity Quantity Quantity Quantity Quantity Quantity Quantity 8.Glycerin 5 5 5 5 5 5 5 5 5 9.propylene glycol 4 4 4 4 4 4 4 4 4 10.triethylamine One One One One One One One One One 11.Sodium 윰 polyacrylate 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 12.Purified water To 100 To 100 To 100 To 100 To 100 To 100 To 100 To 100 To 100 13.Takus Callus Shreddings (Example 1) 5 - - - - - - - - 14.Cellwood Callus Shredding (Example 2) - 5 - - - - - - - 15.Takus callus culture medium (Example 3) - - 5 - - - - - - 16.Cherrywood callus culture (Example 4) - - - 5 - - - - - 17. Crushed Cells Differentiated from Callus of Tak to Root (Example 5) - - - - 5 - - - - 18. Crushed Cells Differentiated to the Roots from the Callus of Cucumber (Example 6) - - - - - 5 - - - 19.Cultural Solution of Cells Differentiated from Callus of Root Tree to Roots (Example 7) - - - - - - 5 - - 20.Cultural Solution of Cells Differentiated from Callus of Roots to Roots (Example 8) 5 - 21.Carbomer Quantity Quantity Quantity Quantity Quantity Quantity Quantity Quantity Quantity

The components 1-7 were melt | dissolved by heating at 65-75 degreeC, and the oily part was manufactured. The water component was prepared by heating and dissolving the components 8-12 at 65-75 degreeC. The prepared oil phase part and component 21 were slowly added while stirring the prepared aqueous part, and emulsified at 7000 to 8000 rpm for 5 minutes to prepare a cream formulation. After cooling to 45 ° C., one of the components 13 to 20 was added and dispersed for 1 minute at 1500 rpm to prepare Examples 1 to 8. In the case of Comparative Example 1, no additional component was added.

Experimental Example 6 Pigmentation Inhibition Effect

A site of 2 × 2 cm ² was set in the lower arms of both arms of the test subjects composed of 20 healthy men and women. Put aluminum foil on the arm so that ultraviolet rays are irradiated only on the test target area, and at the same time, at the distance of 10 cm, two FL20S BLB lamps and FL20S E-30 lamps manufactured by Toshiba Japan at the same time, 0.8 × 10 ^-7 erg / cm ³ / circuit 1 Three consecutive surveys were conducted once daily. The site to be tested was well washed with 70% isopropyl alcohol solution before irradiation. After the irradiation, the formulations of Examples 1 to 8 and Comparative Example 1, which contained a mulberry plant in the UV irradiation site, were applied three times a day. After 3 weeks of application, the degree of pigmentation was visually determined, and the degree of suppression of pigmentation compared to the comparative example was evaluated in three steps of remarkable effect, effectiveness, and invalidity. The results are shown in Table 7.

Applicator Respondents Remarkable effect available invalidity Example 1 2 10 8 Example 2 One 11 8 Example 3 3 6 11 Example 4 2 9 9 Example 5 4 15 One Example 6 3 11 6 Example 7 4 13 3 Example 8 3 10 7 Comparative Example 1 0 7 13

As can be seen from the results of Table 7, Examples 1 to 8 compared to Comparative Example 1 had a large number of people evaluated as a remarkable effect or effective step. Thus, callus lysate of mulberry and coriander, callus culture of mulberry and coriander, lysate of cells differentiated into root in callus of mulberry and coriander, culture of cells differentiated from roots in callus of mulberry and coriander are excellent pigments It was confirmed that it had a deposition inhibitory effect.

Callus cultivars of mulberry and coriander with excellent pigmentation inhibitory effect as above, callus cultures of mulberry and coriander, calligraphy of cells differentiated from roots of callus of mulberry and coriander, callus of mulberry and coriander The culture of differentiated cells has a skin whitening effect, improves skin tone, and prevents or improves aging. These can be mixed and used 2 or more types.

Experimental Example 7 Evaluation of Skin Barrier Function Restoration Effect

In order to measure the skin barrier function recovery effect of Examples 1 to 6, acetone was repeatedly applied to the skin of a hairless mouse to impair barrier function, and then epidermal water loss (TEWL) was measured. When measured by servomed evaporimeter EP1 and reached 4.0 g / m 2 / h, Examples 1 to 6 and Comparative Example 1 were applied to an area of 5 cm 2, and epidermal moisture loss after 1, 2, 4 and 8 hours. The extent to which barrier function is restored is assessed by assessing the extent to which reduction is achieved by measuring. At this time, a conventional lipid mixture (2: 1: 1 mixture of ceramide: cholesterol: fatty acid) was used as a positive control. The epidermal moisture loss at the initial damaged barrier state was set to 100% and the change over time was compared, and the results are shown in Table 8 below.

sample Epidermal moisture loss 1 hour later 2 hours later 4 hours later 8 hours later Example 1 57 47 35 29 Example 2 68 60 42 34 Example 3 65 51 40 32 Example 4 62 57 41 31 Example 5 69 52 47 33 Example 6 69 54 50 33 Comparative Example 1 76 67 61 58 Lipid mixture 65 46 35 26

As can be seen from the results of Table 8, Examples 1 to 6 containing the callus lysate of the mulberry and coriander, the callus culture of the mulberry and coriander, the lysate of cells differentiated into roots in the callus of the mulberry and coriander The amount of epidermal water loss was lower than that of Comparative Example 1 containing no active ingredient and the lipid mixture which is a positive control. Therefore, it was confirmed that the callus lysate of mulberry and coriander, the callus culture of the mulberry and coriander, and the lysate of cells differentiated to the root from the callus of mulberry and coriander have an excellent skin barrier function restoring effect.

Callus lysate of mulberry and coriander, callus culture of mulberry and coriander, and lysate of cells differentiated to roots from callus of mulberry and cotyledon have these effects, and can further prevent and improve aging. . These can be mixed and used 2 or more types.

Experimental Example 8 Evaluation of Skin Tone Improvement Effect

108 adult men and women who were considered to need skin whitening were divided into 9 sets, each of which provided Examples 1 to 8 and Comparative Example 1 and was applied to the face twice a day for 4 weeks.

After using it for 4 weeks, the evaluation of skin tone improvement effect was conducted through a questionnaire about usability and the results are shown in Table 9 below.

Applicator Respondents Very good Good usually Inadequate Example 1 5 6 One 0 Example 2 4 7 3 0 Example 3 3 6 3 0 Example 4 4 4 4 0 Example 5 2 4 5 0 Example 6 3 4 5 0 Example 7 2 4 6 0 Example 8 3 6 3 0 Comparative Example 1 One 3 7 One

As can be seen from the results of Table 9, the people who applied the Examples 1 to 8 was found to feel that the whitening effect is relatively better than the person who applied the Comparative Example 1.

Thus, the callus culture of mulberry and coriander, the callus culture of mulberry and coriander, the culture of cells differentiated from the callus of mulberry and coriander to the root, and the culture of cells differentiated from the callus of mulberry and coriander to the root, skin whitening effect To improve skin tone and prevent or improve aging. These can be mixed and used 2 or more types.

One or more selected from the group consisting of callus lysate, callus culture medium, callus extract, callus extract, lysate of cells differentiated from callus to root, culture solution of cells differentiated from callus and extract of cells differentiated from callus according to the present invention Formulation examples of the cosmetic composition and pharmaceutical composition comprising a composition containing as an active ingredient will be described in more detail, but the cosmetic composition and the pharmaceutical composition is applicable in various formulations, which is not intended to limit the present invention only specific To explain.

[Formulation Example 1] Suspension emulsion lotion

To prepare a suspension emulsion lotion in the composition of Table 10.

Ingredient Content (% by weight) Methylparaben
Spices
Cetyloctanoate
Polyoxyethylene Cured Castor Oil
Polysiloxane
Tocopherol Acetate
ethanol
glycerin
Propylene glycol
Carboxy Vinyl Polymer
Paper mulberry callus shreds
Purified water Suitable amount
Suitable amount
1.0
0.6
0.4
0.1
12.0
6.0
2.0
0.12
2.0
TO 100

Formulation Example 2 Essence

To prepare an essence in the composition of Table 11.

Ingredient Content (% by weight) Methylparaben
Spices
Cetyloctanoate
Octyldodes-16
Coles-24 / Setes-24
Polysiloxane
Tocopherol Acetate
ethanol
glycerin
Xanthan Gum
Propylene glycol
Carboxy Vinyl Polymer
Paper mulberry callus
Purified water Suitable amount
Suitable amount
1.0
0.5
0.2
0.4
0.1
6.0
15.0
0.07
4.0
0.12
6.0
TO 100

Formulation Example 3 Nutrition Cream

To the nutrition cream was prepared in the composition of Table 12.

Ingredient Content (% by weight) Liquid paraffin
Cetostearyl alcohol
Beeswax
Polysiloxane
Lipophilic monostearic acid stearate
Stearic acid
Cetearylglucoside
Monostearic acid polyoxyethylene sorbitan (20 EO)
Hydrogenated Dressin
Cetyloctanoate
Methylparaben
Propylparaben
Spices
Sodium ethyleneamine tetraacetate
glycerin
Carboxy Vinyl Polymer
Propylene glycol
Cucumber Callus Shreds
Purified water 15.0
2.0
3.0
0.5
2.0
2.0
1.5
1.3

2.0
1.0
0.2
0.1
Suitable amount
0.02
5.0
0.10
2.0
2.0
TO 100

Formulation Example 4 Gel

To prepare a gel in the composition of Table 13.

Ingredient Content (% by weight) antiseptic
Spices
Cetearylglucoside
Cetyloctanoate
glycerin
Jojoba wax
Dipantenol
ethanol
Carboxy Vinyl Polymer
Propylene glycol
Culture of Differentiated Cells from Roots of Callus of Paper mulberry
Purified water Suitable amount
Suitable amount
1.5
1.0
5.0
3.0
1.0
7.0
0.6
3.0
6.0

TO 100

[Formulation Example 5] Pack

To prepare a pack in the composition of Table 14.

Ingredient Content (% by weight) Methylparaben
Spices
Cetyloctanoate
Polyoxyethylene Cured Castor Oil
Polysiloxane
Cellulose gum
ethanol
glycerin
Propylene glycol
Polyvinyl alcohol
Crushed Cells Differentiated from Roots of Callus to the Roots
Purified water Suitable amount
Suitable amount
1.0
0.6
0.2
0.2
6.0
5.0
3.0
15.0
3.0

TO 100

[Formulation Example 6] Ointment in external skin preparations

The ointment was prepared in a conventional manner according to the composition described in Table 15 below.

ingredient Content (% by weight) Purified water Balance glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 15.0 Beta Glucan 7.0 Carbomer 0.1 Culture of Differentiated Cells from Roots of Callus of Paper mulberry 1.0 Caprylic / Capric Triglycerides 3.0 Squalane 1.0 Cetearyl Glucoside 1.5 Sorbitan stearate 0.4 Cetearyl Alcohol 1.0 antiseptic Quantity incense Quantity Pigment Quantity Beeswax 4.0

1 is a graph of the results of Experimental Example 5 for the melanin production inhibition rate (%) of the composition according to the present invention.

Claims (9)

Skin containing one or more selected from the group consisting of callus lysate of callus, callus culture, callus extract, lysate of cells differentiated from callus, cultures of cells differentiated from callus and extracts of cells differentiated from callus as active ingredients Regeneration promoting composition. Skin containing one or more selected from the group consisting of callus lysate of callus, callus culture, callus extract, lysate of cells differentiated from callus, cultures of cells differentiated from callus and extracts of cells differentiated from callus as active ingredients Anti-wrinkle or improvement composition. Skin containing one or more selected from the group consisting of callus lysate of callus, callus culture, callus extract, lysate of cells differentiated from callus, cultures of cells differentiated from callus and extracts of cells differentiated from callus as active ingredients Whitening composition. Blood containing one or more selected from the group consisting of callus lysate of callus, callus culture, callus extract, lysate of cells differentiated from callus, cultures of cells differentiated from callus and extracts of cells differentiated from callus as active ingredients Secondary moisturizing composition. Skin containing one or more selected from the group consisting of callus lysate of callus, callus culture, callus extract, lysate of cells differentiated from callus, cultures of cells differentiated from callus and extracts of cells differentiated from callus as active ingredients Tone improving composition. Aging containing at least one selected from the group consisting of callus lysate of callus, callus culture, callus extract, lysate of cells differentiated from callus, cultures of cells differentiated from callus and extracts of cells differentiated from callus as active ingredients Prevention or improvement composition. 7. The method according to any one of claims 1 to 6, The mulberry plant is called Broussonetia kazinoki ) or Coriander ( Broussonetia) papyrifera ). 7. The method according to any one of claims 1 to 6, The cell differentiated in callus comprises a cell differentiated from callus to leaf, stem, or root. 7. The method according to any one of claims 1 to 6, The composition comprises the total composition weight of at least one selected from the group consisting of callus lysate, callus culture medium, callus extract, lysate of cells differentiated from callus, culture solution of cells differentiated from callus and extracts of cells differentiated from callus It comprises 0.0001 to 25.0% by weight relative to the composition.

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