KR20130023850A - Cosmetic composition comprising nano capsule containing extract of camellia sinensis, curcuma longa, magnolia obovata and aralia continentalis and manufacturing method thereof - Google Patents

Abstract

PURPOSE: A cosmetic composition containing a nanocapsule of a Camellia sinensis extract, a Curcuma longa extract, a Magnolia obovata extract, and an Aralia continentalis extract is provided to quickly penetrate the skin with an active ingredient. CONSTITUTION: A cosmetic composition contains a nanocapsule containing a Camellia sinensis extract, a Curcuma longa extract, a Magnolia obovata extract, and an Aralia continentalis extract. The green tea extract contains epigallocatechin gallate(EGCG). The Curcuma longa extract contains curcumin, demethoxycurcumin, and bisdemethoxycurcumin. The Magnolia obovata extract contains honokiol and 4-methoxyhonokiol. The Aralia continentalis extract contains continentalic acid and kaurenoic acid.

Description

Cosmetic composition comprising nanocapsules of green tea extract, turmeric extract, thick extract and poison extract, and a method for preparing the same {Cosmetic Composition Comprising Nano Capsule Containing Extract of Camellia Sinensis, Curcuma Longa, Magnolia Obovata and Aralia Continentalis and Manufacturing Method Thereof}

The present invention relates to a cosmetic composition and a method for producing the same, and more particularly, the present invention relates to a cosmetic composition comprising a nanocapsules of green tea extract, turmeric extract, thick extract and poison extract.

Aging is a functional, structural, and biochemical process that occurs continuously from birth to death, and occurs throughout the cells and tissues that make up the human body. It shows metabolic rate, increased disease, and decreased adaptability. This is a phenomenon that leads to the death of the whole body.

Among these, skin aging is largely divided into two studies. One is 'internal aging' and the aging phenomenon according to the age increase, and the other is 'external aging' and aging phenomenon due to external environment such as ultraviolet rays or pollution. Many theories have been suggested so far about skin aging, but the most scientific approach to skin aging is skin aging by oxidation. Human skin is composed of the epidermis, the dermis, and connective tissue including the stratum corneum, of which the dead layer consists of dead cell layers formed through the differentiation of keratinocytes, the basal cells of the epidermis. It plays a role of protecting the human body from the influence of the external environment. In addition, the dermal layer existing inside the skin is composed of collagen (collagen) and elastin (elastin) to play a role in protecting the skin from sagging by giving elasticity of the skin. The theory of antioxidant is the theory that collagen and elastin are damaged by free radicals generated by factors such as external ultraviolet rays, and accordingly, the elasticity of the skin is reduced and wrinkles are generated.

Until now, natural antioxidants such as vitamin E, vitamin C, carotenoids, glutathione and plant extracts have been used to prevent the aging of the skin, especially wrinkles. Causes irritation, redness, and inflammation, which have serious problems with the skin.

Accordingly, the present inventors have tried to develop a cosmetic with excellent anti-wrinkle effect and antioxidant effect, the first time to verify the anti-aging and wrinkle improvement efficacy and efficacy unknown in the domestic herbal medicines such as green tea, turmeric, thick and live alone. In addition, in order to solve the problems of color and odor affecting skin irritation and cosmetic formulations that herbal medicines generally have, nanotechnology is applied to develop a stable, irritating and effective functional cosmetic.

In this regard, Korean Patent Laid-Open Publication No. 10-2010-0054026 describes a skin external preparation composition containing an extract of Tofu Park as an active ingredient, and Korean Patent No. 10-0902256 describes a mixed turmeric composition comprising turmeric extract. It describes the use of liver protection and blood pressure lowering, and Korean Patent No. 10-0825432 describes the anti-inflammatory and anti-allergic use of the venom extract, but none of the prior literature Wrinkle improvement and antioxidant efficacy, and even cosmetic compositions made thereof as a nano formulation is not disclosed.

It is an object of the present invention to establish a delivery system capable of selectively delivering skin active ingredients to the skin by nanoencapsulating green tea, turmeric, thick and toxin extract having excellent effects on wrinkle improvement and antioxidant, and the nanoencapsulated cosmetic composition and It is to provide a preparation method thereof.

As one aspect for achieving the above object, the present invention green tea ( Camellia sinensis ) extract, turmeric ( Curcuma) longa extract, Magnolia obovata extract and poisonous flower ( Aralia) continentalis ) relates to a cosmetic composition for anti-wrinkle and antioxidant, including nanocapsules encapsulated extract.

Preferably, the green tea extract comprises Epigallocatechin gallate (EGCG).

Also preferably, the turmeric extract includes curcumin, dimethoxycurcumin, and bismethoxycurcumin.

Also preferably, the thick gourd extract includes Honokiol and 4-Methoxyhonokiol.

Also preferably, the poison extract contains Continentalic acid and Kaurenoic acid.

In addition, the cosmetic composition includes one formulation selected from the group consisting of softening cream, nourishing cream, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray and powder.

As another aspect, the present invention relates to a method for producing a cosmetic composition for improving wrinkles and antioxidants, including nanocapsules in which the green tea extract, turmeric extract, hul extract extract and poison extract are encapsulated.

Preferably, the manufacturing method

(1) preparing a green tea extract, turmeric extract, pak extract and poison extract; And

(2) nano-encapsulating the green tea extract, turmeric extract, thick extract and poison extract.

Also preferably, the step (2)

Dissolving the green tea extract, turmeric extract, hulhu extract and poison extract and biodegradable polymer in a volatile organic solvent;

Dispersing and stirring the solution in water to form particles; And

And evaporating off the volatile organic solvent in the particle solution.

The cosmetic composition using the green tea, turmeric, scabbard and poison extract of the present invention has an excellent effect on improving wrinkles and anti-aging, and unlike the conventional plant extract-containing functional cosmetics simply using extracts, the present invention is active through nano-processing. The ingredients can penetrate deep into the skin quickly. Therefore, the present invention is more excellent in supplying the active ingredient to the skin tissue than conventional herbal cosmetics, it can be maintained in the active ingredient as long-term storage, it is possible to provide a functional cosmetic with a stable, irritation-free and increased effectiveness .

1 shows the DHHP scavenging ability of compounds. In the graphs VitC 1, 2, 3 represent 50, 200 and 1000 uM, respectively, EGCG, RV (resveratrol), BDMC (bisdimethoxycurcumin), DMC (dimethoxycurcumin), CUR (curcumin), honokiol (honokiol) ) And 4-MHK (4-methoxyhonokiol) 1, 2, 3 represent 0.5, 5, 50 uM, respectively, CA (continental acid) and KA (kaurenoic acid) 1, 2, 3 are 5, 50 and 250 uM. p <0.05 *, p <0.01 **, p <0.001 ***
Figure 2 shows the results of NF-κBSEAP Reporter gene assay of Transfectant HaCaT cells. BDC (bismethoxycurcumin), DMC (dimethoxycurcumin), CA (continental acid), KA (kaurenoic acid), 4-MHK (4-methoxyhonokiol), ### is p <0.001 vs p <0.01 ** p <0.001 ***.
Figure 3 shows the results of TNF-α activity through reverse transcription polymerase chain reaction (RT-PCR).
Figure 4 shows the results of MMP-1, MMP-2 activity measurement by reverse transcription polymerase chain reaction (RT-PCR).
5 shows MMP-1 expression in human HaCaT cells via Western blot.
6 shows MMP-1 expression in human HaCaT cells via Western blot.
7 shows MMP-1 expression in neonatal foreskin derived fibroblasts via Western blot.
8 shows MMP-1 expression in neonatal foreskin derived fibroblasts via Western blot.
9 shows MMP-1 expression in neonatal foreskin derived fibroblasts via ELISA. In BDMC (bisdimethoxycurcumin), DMC (dimethoxycurcumin), CUR (curcumin) 1, 2, 3 represent 0.15, 1.5, 15 uM, respectively, RV (resveratrol), honokiol and 4-MHK (4- 1, 2, 3 represent 0.1, 1, 10 uM, respectively, in methoxy honokiol), and 1, 2, 3 represent 1, 10, 50 uM of CA (continental acid) and KA (kaurenoic acid), respectively. . # Represents p <0.05 versus p <0.05 *, p <0.01 **, p <0.001 ***.

In the present invention, the term "green tea ( Camellia) sinensis ) "refers to an evergreen broad-leaved shrub belonging to the genus Tea, and preferably, the green tea extract comprises Epigallocatechin gallate (EGCG).

As used herein, the term "turmeric (Curcuma longa ) "refers to perennial grasses of Gingeraceae, also called turmeric. Preferably, the turmeric extract includes Curcumin, Demethoxycurcumin and Bismethoxymethoxycucumin.

As used herein, the term "Magnolia (Magnolia obovata ) "refers to a deciduous tree belonging to the Magnolia family, and preferably, the gourd extract includes Honokiol and 4-Methoxyhonokiol.

In the present invention, the term " Aralia continentalis ) "is a perennial herb that is included in the family Arboraceae, also called dulcis, durdu, auditorium, and vigor, and young sprouts are used for food, and roots are used for medicinal purposes. Continentalic acid) and Kaurenoic acid.

The green tea, turmeric, scabbard and poison extract of the present invention are cut to about 1 to 20 times the weight (kg), preferably about 3 times, by cutting out dried plants, roots or ground parts (leaves, stems) and the like. 10 to 10 times water, C1 to C4 lower alcohol or a mixed solvent thereof, preferably ethanol, about 1 to 24 hours at an extraction temperature of 0 ° C to 100 ° C, preferably 10 ° C to 80 ° C, preferably For about 2 hours to 5 hours can be obtained by filtration, concentration under reduced pressure or drying the extract obtained by using a cold, hot water extraction, ultrasonic extraction, reflux cooling extraction method. It may also be obtained by performing a conventional fractionation process (Harborne J.B., Phytochemical methods: A guide to modern techniques of plant analysis, 3rd Ed. Pp 6-7, 1998).

In the cosmetic composition of the present invention, the green tea, turmeric, grommets, and poison extracts are 0.00001 to 15.0% by weight, more preferably 0.0001 to 10% by weight, most preferably 0.0001 to 5% by weight, based on the total weight of the composition. May be included. When the weight of these active ingredient extract is less than 0.00001% by weight, the effect is less likely to appear, when it exceeds 15.0% by weight, the increase in effect due to the increase of the content is weak and there is a problem that the stability of the formulation is not secured.

The cosmetic composition of the present invention is characterized in that the herbal extracts of the present invention are encapsulated in nano vesicles and nanoencapsulated. Generally, herbal extract-containing cosmetics are known to have better skin activity than general cosmetics, but they may cause skin irritation, redness, and inflammation, which may have side effects on the skin, and problems of color and smell affecting cosmetic formulations. Has been raised.

Thus, in the present invention, in order to solve the skin stability and skin absorption problem of the herbal extract raw material itself and to provide a high-quality cosmetics, by applying nanotechnology to provide a functional cosmetic material that is more stable, less irritating and effective than conventional bio materials do.

Encapsulation technology refers to the technology of encapsulating solid, liquid, or gaseous substances or packaging inside a substance or tissue to release its contents at a controlled rate under certain conditions. It is called (nanocapsule). Nanocapsules can be defined as several nanometers (nm) to several hundred nanometers. Preferably, in the present invention by applying a nano-spheres manufacturing technology of 50 to 500 nm size belonging to the nano range to increase the percutaneous absorption efficiency of the functional cosmetics.

The cosmetic composition of the present invention can be used for wrinkle improvement and antioxidant.

As used herein, the term 'antioxidative capacity' means a function of inhibiting oxidation of skin components by highly reactive free radicals and free radicals under the influence of ultraviolet rays. Free radicals and free radicals oxidize the constituents of the skin to produce peroxides, resulting in structural and functional damage to the skin to promote aging, and the antioxidant function of the present invention serves to protect the skin from it. . The cosmetic composition of the present invention may perform a function of inactivating the free radicals.

Therefore, in a specific embodiment, the present inventors evaluated antioxidant efficacy by evaluating the efficacy of DPPH (1,1-diphenyl-2-picryl hydrazyl) by the active ingredients included in green tea, turmeric, humbac and poison extract in relation to antioxidant activity. It was confirmed (Fig. 1).

As used herein, the term 'wrinkle improvement' means to maintain or enhance the ability to be associated with wrinkles and elasticity of the skin. Collagen and elastin present in the dermal layer of the structure of the skin controls the elasticity of the skin, collagen biosynthesis is affected by the internal and external skin. Specifically, as an internal factor, cellular activity is decreased due to natural aging, collagen fiber decreases, and as an external factor, active oxygen generated due to excessive irradiation and stress of ultraviolet light is a thiol group of protein. It increases the expression of collagenase, which is an enzyme (Matrix Metalloproteinases-1 (MMP-1)) that inhibits the activity of enzymes or breaks down collagen, elastin, etc. Cause results.

In particular, the synthesis and degradation of extracellular matrix in vivo is properly coordinated, but as aging progresses, its synthesis decreases and matrix metalloproteinase (MMP), an enzyme that degrades collagen, especially MMP-1, MMP. -2, MMP-13 is promoted, the elasticity of the skin is reduced, wrinkles are formed.

Accordingly, the present inventors have shown that, in a specific embodiment, with respect to wrinkle improvement, the active ingredients included in green tea, turmeric, hul-bak and toxin extracts inhibit the expression of MMP-1, MMP-2, MMP-13 in human keratinocytes. In addition, the inhibitory effect on the activation of NF-κB (ultraviolet or TNF-α.IL-1) in human keratinocytes and the inhibition of MMP activity in fibroblasts derived from human neonatal foreskin (FIG. 2 to 2) were evaluated. 9).

As used herein, the term 'cosmetic composition' is a composition comprising the extract, the formulation may be in any form. Examples of such formulations are cosmetics prepared using the cosmetic composition, such as creams, packs, lotions, essences, lotions, foundations and makeup bases, and any form of these formulations to achieve the object of the present invention. Furnace can also be produced and commercialized, but is not limited to the above examples. Preferably, the cosmetic composition may include one formulation selected from the group consisting of softening cream, nourishing cream, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray and powder have.

In addition, the cosmetic composition according to the present invention may optionally add purified water, humectant, higher alcohol, higher fatty acid, emollient, chelating agent, thickener, permanent ingredient, pH adjusting agent, flavoring agent, etc. according to the formulation or purpose of use of the cosmetic. have.

Method for producing a cosmetic composition comprising a nanocapsule containing the herbal extract of the present invention, preferably,

(1) preparing a green tea extract, turmeric extract, pak extract and poison extract; And

(2) encapsulating the green tea extract, turmeric extract, thick extract and poison extract in nano vesicles to nanoencapsulate.

The step of preparing the green tea extract, turmeric extract, halved extract and poison extract, about 1 to 20 times the weight (kg) by cutting the dried outpost, root or ground portion (leaf, stem), etc., preferably Preferably about 3 to 10 times water, C1 to C4 lower alcohol, or a mixed solvent thereof, preferably ethanol, from about 1 hour at an extraction temperature of 0 ° C to 100 ° C, preferably 10 ° C to 80 ° C. For 24 hours, preferably about 2 to 5 hours, the extract obtained by using a cold, hot water extraction, ultrasonic extraction, reflux cooling extraction method and the like can be obtained by filtration, concentrated under reduced pressure or dried. It may also be obtained by carrying out a conventional fractionation process.

Next, the step of encapsulating the nano-encapsulated green tea extract, turmeric extract, thick extract and poison extract can be carried out.

As a specific preferred embodiment of nanoencapsulation, a solvent evaporation method, a solvent displacement method or an emulsification solvent diffusion method, a supercritical fluid extraction process and the like can be used.

The solvent evaporation method dissolves a substance encapsulated in a volatile organic solvent and an oil-soluble substance in water, disperses the solution in an aqueous solution containing an anti-agglomerating agent to form particles, and distills the volatile organic solvent to prepare particles. will be. At this time, the size of the particles largely depends on the stirring speed at the time of dispersing, and when using a typical agitator, particles of several microns are formed, and when using a high speed stirring homogenizer, hundreds of nanometers are used. Particles may be formed. In general, in order to obtain particles of several tens of nanometers, particles having a size of 100 nanometers or less may be obtained by using a high pressure homogenizer. When the solvent evaporation method is used, the organic solvent uses a highly volatile solvent, and specific examples thereof include methylene chloride and ethyl acetate.

Solvent replacement or emulsification solvent diffusion method dissolves the encapsulating material and oil-soluble material in the volatile organic solvent mixed with water, dispersing the solution in an aqueous solution containing an anti-flocculation agent to form particles, and distilling the volatile organic solvent to distill the particles. To prepare. Since this method uses a volatile organic solvent mixed with water, particles are formed by the difference in the solubility of the encapsulating material and the useful material in the cosolvent of water and the organic solvent. Therefore, water-soluble surfactants and other surfactants for preventing aggregation It is preferable to include. In addition, hydrophobic surfactants may be used to stabilize the encapsulated oil soluble actives. Factors affecting the size of the particles produced by the solvent replacement method is the ratio of the organic solvent and water, the concentration of the encapsulating agent and sparingly soluble active ingredient used in the organic solvent used, the amount of hydrophobic surfactant, The amount of hydrophilic surfactant, and the stirring speed, and in general, the size of particles obtained through these components and methods is between 100 and 300 nm.

Supercritical fluid extraction process can be widely used in medicine, food, cosmetics, etc., because it has no toxic solvent or surfactant in the particles, compared to other extraction processes, RESS (Rapid Expansion of Supercritical Solutions), GAS (Gas Anti-Solvent), Supercritical fluid Anti-Solvent (SAS), Aerosol Solvent Extraction System (ASES), Solution Enhanced Dispersion by Supercritical Fluids (SEDS), Particles from Gas Saturated Solutions (RPGSS), and Reactive Precipitaion in Supercritical Solution (RPSS) methods can be used.

 As a preferred embodiment, the step of encapsulating the nano-encapsulation by encapsulating the green tea extract, turmeric extract, pak bak extract and poison extract in the cosmetic preparation method of the present invention,

Dissolving the green tea extract, turmeric extract, hulhu extract and poison extract and biodegradable polymer in a volatile organic solvent;

Dispersing and stirring the solution in water to form particles; And

It may include the step of removing by evaporating the volatile organic solvent in the particle solution.

The biodegradable polymer should be excellent in biocompatibility and biodegradability in consideration of the safety of the body when the prepared nanoparticles penetrate the skin, for example, starch, chitosan, alginic acid, lecithin, glucomannan, polysaka as natural polymers. Ride, acacia gum, Irish moss, karaya gum, gum tragacanth, gum guaiac, xanthan gum, locust bean gum and derivatives thereof, including proteins such as casein, gelatin, collagen, albumin, globulin and fibrin, cellulose, Naturally derived carbohydrates and derivatives thereof, such as dextrin, pectin, starch, agar, and mannan, as synthetic polymers, polyvinyl polymers and derivatives thereof (e.g. polyvinylpyrrolidone, polyvinyl alcohol, polyvinyl methyl ether, polyvinyl) Ethers), polycarboxylic acids and derivatives thereof (e.g. polyacrylic acid, polymethacrylic acid, polymethylmethacrylate, etc.), poly Hydrocarbones such as lene and polypropylene, isomers thereof, polysaccharides and derivatives thereof (polysucrose, polyglucose, polylactose and salts thereof), and the like, and polymers in which the aforementioned polymers are crosslinked are possible. . Naturally-derived polymers are known to have excellent biocompatibility, and the polymers described above may be applied to the nanoparticles of the present invention as polymers having excellent biocompatibility.

Also, for example, fatty acid polymers (e.g., polylactic acid, polyglycolic acid, polycitric acid, polymaline acid, etc.) and derivatives thereof, poly-, alpha-, cyanoacrylic acid and derivatives thereof, poly-, beta-, Hydroxybutyric acid, polyalkylene oxalate (e.g. polymethylene oxalate, polytetramethylene oxalate, etc.), polyorthoesters, polyorthocarbonates, polycarbonates (e.g. polyethylene carbonate, polyethylene-propylene carbonate Etc.), and polyamino acids (e.g., polygammabenzylglutamic acid, polyalanine, polygammamethylglutamic acid, etc.) can be used, and polystyrene, polyacrylic acid and its derivatives, polymethacrylic acid and derivatives thereof having excellent biocompatibility. , Copolymers of acrylic acid-methacrylic acid, polyamides (eg nylon), polyethylene terephthalate, polyamino acids, silicone high Monopolymers such as porcelain, dextranstearate, ethylcellulose, acetylcellulose, nitrocellulose, polyurethane, maleic anhydride copolymer, ethylene-vinylacetate copolymer, polyvinylacetate, polyvinyl alcohol, polyacrylamide Polymers, two or more copolymers, and mixtures of these polymers are available, and derivatives and salts thereof may be used.

The volatile organic solvents include methanol, ethanol, acetone, THF (tetrahydrofuran), acetonitrile, and 1,4-dioxane as volatile organic solvents mixed with water. And volatile organic solvents that do not mix with water. Halogenated alkanes (chloromethane, dichloromethane, chloroform, tetrachloromethane, dichloroethane, etc.), ethyl acetate, diethyl ether, cyclohexane, benzene, toluene, etc. Can be illustrated.

In step (2), an emulsifier can be added, and as an emulsifier, an emulsifier of polyoxyethylene type, saturated and unsaturated alkyl sulfates, ammonium of fatty acids, alkanesulfonates, fatty alcohol phosphorates, fatty acid partial glycerides, polyglycerol esters , Propylene glycol esters of fatty acids, lactates of fatty acids, and the like.

In addition, a microfluidizer (Microfluidics Corporation, USA) which uses a high pressure and high shear force to make an emulsion into uniform nano-sized particles may be used to manufacture nanoparticles, and the pressure may be between 500 bar and 1500 bar. The flow rate can be adjusted between 20 ml / min and 150 ml / min.

Hereinafter, the present invention will be described in more detail. However, the following examples are illustrative of the present invention, and the present invention is not limited by the following examples.

Example  1. Preparation of experimental materials

1-1. Compound and Laboratory Equipment

Green tea ( Camellia Epigallocatechin gallate (EGCG) isolated from sinensis ), Curcumin, Demethoxycurcumin, Bismethoxycurcumin and Bismethoxycurcumin, Magnolia isolated from Curcuma longa Honokiol and 4-Methoxyhonokiol isolated from obovata ), Continentalic acid and Kaurenoic acid isolated from Aralia continentalis were tested.

As reagents and analyzers, Dulbecco's phosphate buffered saline (D-PBS), Dulbecco's modified Eagle's medium (DMEM), penicillin-streptomycin mixed solution, sulfanilamide, naphtylethylenediamine dihydrochloride (naphtylethylenediamine dihydrochloride) The back was purchased from Sigma (St Louis, MO), the fetal bovine serum (FBS) was purchased from South Pacific (New Zealand), and the geneticin (antibiotic G-418) was purchased from Gibco BRL (Grand Island, NY, USA). Was used. Related culture dishes were purchased from SPL (Korea) company's animal cell culture titration product.

As an experimental device, BIO-LINK crosslinker (Vilber Lourmat) Ultra violet spectrophotometer (Jasco), Microplate reader (Molecular Device), Microfluorometer (Molecular Device), LAS-1000 (Fuji) were used.

1-2. Cell culture

Human keratinocyte (HaCaT) cells and fibroblas derived from human neonatal foreskin are cultured in an incubator maintained at 37 ° C., 5% CO ₂ under DMEM containing 10% FBS and 1% antibiotic (penicillinstreptomycin). It was. Transfectant HaCaT cells infected with pNF-kB SEAP NPT were further selected with 500 μg / ml of neomycin derivative geneticin.

Example  2. DPPH  Antioxidant efficacy measurement

DPPH (1,1-dimethyl-2-picrylhydrazyl radical) scavenging activity was measured to measure the antioxidant activity of the sample. The DPPH solution was dissolved in methanol at a concentration of 1.5 × 10 ^-4 M and L-ascorbic acid (vitamin C) was used as a control. After treating DPPH solution to various concentrations of samples, the reaction was performed at room temperature for 30 minutes, and the absorbance was measured at 540 nm.

Example  3. Transfectant HaCaT  Cell NF -KB SEAP  Reporter gene Assay

HaCaT cells were cultured at a 24-well plate cell concentration of 1 × 10 ⁵ cells / well and treated with sample compounds. Then, 50 μl from each well was reacted for 5 minutes at 65 ° C. to inactivate the enzyme, stored at 20 ° C., and 25 μl of each sample was placed in 3 repeats in a 96 well plate and 100 μl of assay buffer (500 μM 4methylumbelliferyl phosphate). , 1 mM MgCl 2, 2M diethanol amine) was reacted at 37 ° C. for 60 minutes, and after 360 nm excitation, the fluorescence emitted at 449 nm was measured, and the alkaline phosphatase secreted due to the activation of NF-κB. The activity of was measured (Fig. 2).

Example  4. Reverse transcription Polymerase chain reaction (RT-PCR)  through TNF -α, MMP -One, MMP -2 activity measurement

Cell suspensions of HaCaT and neonatal foreskin-derived fibroblasts were divided into 12 well plates at a cell concentration of 2 × 10 ⁵ cells / well, stabilized by incubation for 24 hours, and then treated with a concentration of candidate compounds for 12 hours. treatment). Thereafter, the culture medium of each well was removed and UV B 30mJ / cm ² was irradiated with D-PBS in the case of HaCaT cell line, and TNF-α was treated in the concentration of 10 ng / ml in the case of fibroblast line, and then pretreated again. Each test compound was retreated under the same conditions and further incubated for 5 hours. After the culture was removed, total RNAs were extracted using Easy-BLUE (IntronBioTech, Korea), and 260/280 nm was obtained using UV-spectrophotometery. 5 μg of each sample was taken and cDNA was synthesized using reverse transcriptase. Thereafter, the synthetic products were serially synthesized through cDNA based PCR to obtain PCR products such as MMP-1, MMP-2, and TNF-α. The same amount of the PCR product of each sample was electrophoresed on agarose gel to isolate the amplified gene, and then stained with ethidium bromide, photographed with a UV illumonator-image analyzer, and analyzed. GAPDH was used as a loading control (FIGS. 3 and 4).

Example  5. Western Blat  through MMP -One, MMP Confirmation of -13 Expression

Western blot was performed to determine the effect of each compound on MMP-1, MMP-13 gene expression. Cell suspensions of HaCaT cells and neonatal foreskin-derived fibroblasts were divided into 6-well plates at a cell concentration of 3 X 10 ⁵ cells / well and incubated for 24 hours to stabilize, followed by treatment of each candidate compound group for 24 hours. Pre-treatment. Thereafter, the culture medium of each well was removed and UV B 30 mJ / cm ² was irradiated with D-PBS in the case of HaCaT cell line, and TNF-α was treated in the case of fibroblast line at a concentration of 10 ng / ml and then again. Each experimental compound was retreated and further incubated for 24 hours under the same conditions as the pretreatment. After completion of the experiment, lysis buffer (20 mM Tris-Cl pH 7.5, 150 mM NaCl, 1 mM Na2EDTA, 1 mM EGTA, 1% NP-40, 1 mM DTT, 1 mM Na3VO4, 1 mM PMSF and protein inhibition cocktail) was added. Total protein was separated by treatment, and the amount of protein in all samples was quantified using UV spectrophotometery 595 nm. Load and fractionate 20 μg of total protein in 10% SDS-PAG and induce blocking, gel-membrane transfer, primary and secondary antibody reactions using a conventional western blot method, and use WEST-Save as an ECL-like reagent. (AB Fronteer, Korea) was recorded by comparing the amount of fluorescence developed on the membrane with the final LAS1000 (Figs. 5 to 8).

Example  6. ELISA  through MMP -1 expression confirmation

MMP-1 secreted into the cell culture was measured using a RayBioHuman MMP-1 ELISA Kit (Cat #: ELH-MMP1-001). Fibroblasts were treated after TNF-α treatment for 24 hours, followed by dilution. Plate-diluted medium coated with anti-human MMP-1 was reacted at 4 ° C., followed by reaction with MMP-1 antibody, HRP-Streptavidin, and color development through TMBtetramethylbenzidine), and absorbance at 450 nm was measured (FIG. 9). The amount of expressed human MMP-1 was corrected by measuring the total protein extracted from the total cells of each treatment group used in the experiment (Bradford method).

Experiment result

Green tea ( Camellia Epigallocatechin gallate (EGCG) isolated from sinensis showed strong DHHP scavenging activity.

Curcumin, Dimethoxycurcumin and Bismethoxymethoxycurcumin isolated from Curcuma longa showed DHHP scavenging ability, NF-κB inhibition, TNF-α transcription inhibition and MMP in keratinocytes. -1 inhibition, and the only effect in ELISA against MMP-1 secreted from fibroblasts, and is considered to be a natural product showing a strong effect at low concentrations.

Hug ( Magnolia Honokiol and 4-Methoxyhonokiol isolated from obovata ), and Continentalic acid and Kaurenoic acid isolated from Aralia continentalis have DHHP scavenging ability. It was confirmed that keratinoids exhibited weak NF-κB inhibition and TNF-α transcription inhibition and reduced expression of MMP-1 through western blot. (Table 1)

origin
Compound name HaCaT keratinocyte Fibroblast NF-κB TNF-α MMP-1 MMP-1
ELISA
Turmeric Curcumin + ++ ++ Dimethoxycurcumin ++ ++ ++ + Bismethoxycurcumin ++ ++ ++ + Thick Honokiol + - + - 4-methoxy honokiol + - ++ - Solo Continental Acid + - + - Kaureno Iksan + - ++ -

Example  7. Fabrication of Nano Formulations

Using a fatty acid polymer such as polylactic acid, polyglycolic acid, or isopropyl myristate (IPM) as a biodegradable polymer, nanocarriers having a size of 100-200 nm are prepared by solvent diffusion method, and an emulsifier (emulsifier of polyoxyethylene type, saturated and unsaturated Alkyl sulfates, ammoniums of fatty acids, alkanesulfonates, fatty alcohol phosphates, fatty acid partial glycerides, polyglycerol esters, propylene glycol esters of fatty acids, lactates of fatty acids) and then stirred in a high speed stirring homogenizer To prepare a cosmetic formulation, such as a green tea, turmeric, bakbak and solvent cream containing a lubricating extract.

Example  8. Of nano formulation  Evaluation of efficacy related to wrinkle improvement

In 20 healthy women, the control product (physiological saline 1cc) and the nano formulation (1cc) prepared in Example 7 were applied to the subject twice a day (morning, before going to bed) around the eyes, and 4 weeks later. And after 8 weeks, the skin moisturizing degree, elasticity, wrinkle improvement effect, skin irritation degree and feeling of sensory evaluation were to be evaluated, 5 points if the feeling is excellent, 0 points if no effect at all 6 steps each (However, the degree of skin irritation and the feeling of use are 0 points when there is no irritation or the feeling of use is excellent, and 5 points indicate when the irritation is severe or the feeling is bad). The results are shown in Table 2.

Nano formulation of Example 7 Control group After 4 weeks 8 weeks later After 4 weeks 8 weeks later Skin hydration 3.6 4.2 1.2 1.1 Elasticity 2.8 3.1 0.8 0.8 Wrinkle improvement 3.0 4.2 0.9 1.2 Degree of skin irritation 0.0 0.0 0.0 0.0 Feeling 0.0 0.0 0.0 0.0

Although the preferred embodiments of the present invention have been described, the present invention is not limited to the specific embodiments described above. That is, those skilled in the art to which the present invention pertains can make many changes and modifications to the present invention without departing from the spirit and scope of the appended claims, and all such appropriate changes and modifications are possible. Equivalents should be considered to be within the scope of the present invention.

Claims (9)

Green tea ( Camellia sinensis extract, Curcuma longa extract, Magnolia obovata extract and poisonous flower ( Aralia) continentalis ) Wrinkle improvement and antioxidant cosmetic composition comprising a nanocapsule containing an extract.
The cosmetic composition of claim 1, wherein the green tea extract comprises Epigallocatechin gallate (EGCG).
The cosmetic composition of claim 1, wherein the turmeric extract comprises curcumin, dimethoxycurcumin, and bismethoxycurcumin.
The cosmetic composition according to claim 1, wherein the thick gourd extract comprises Honokiol and 4-Methoxyhonokiol.
The cosmetic composition of claim 1, wherein the poison extract comprises Continentalic acid and Kaurenoic acid.
According to claim 1, wherein the nanocapsules have a diameter of 50 to 500 nm cosmetic composition.
According to claim 1, wherein the cosmetic composition is a flexible cosmetics, nourishing cosmetics, nourishing cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray and powder one formulation selected from the group Cosmetic composition to have.
(1) preparing a green tea extract, turmeric extract, pak extract and poison extract; And
(2) comprising the steps of nano-encapsulating the green tea extract, turmeric extract, thick extract and poison extract,
The manufacturing method of the cosmetic composition of any one of Claims 1-6.
The method of claim 8, wherein step (2)
Dissolving the green tea extract, turmeric extract, humbac extract and poison extract and biodegradable polymer in volatile organic solvent;
Dispersing and stirring the solution in water to form particles; And
Method for producing a cosmetic composition comprising the step of removing by evaporating the volatile organic solvent in the particle solution.

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