KR20160020145A - Cosmetic Composition comprising Ribonucleic acid and Extract of Panax ginseng as active ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition containing ribonucleic acid and a Panax ginseng extract as active ingredients. The present invention relates to a cosmetic composition inhibiting an activity of ribonuclease (RNase) promoting the degradation of ribonucleic acid by comprising a Panax ginseng extract and ribonucleic acid. According to the present invention, a formation containing the Panax ginseng extract has increased stability of micro-ribonucleic acid having excellent anti-aging activity and whitening activity, to maximize an effect of the micro-ribonucleic acid. The Panax ginseng extract of the present invention can be effectively used in preservation of the ribonucleic acid in cosmetics including the ribonucleic acid such as micro-ribonucleic acid.

Description

(Cosmetic Composition of Ribonucleic acid and Extract of Panax ginseng as active ingredient)

The present invention relates to a cosmetic composition comprising ribonucleic acid and ginseng extract as an active ingredient. The present invention relates to a cosmetic composition capable of inhibiting the activity of a ribonucleic acid degrading enzyme that promotes degradation of ribonucleic acid by incorporating ginseng extract together with ribonucleic acid.

Ribonucleic acid or RNA is a type of nucleic acid that forms a nucleotide based on ribose, unlike deoxyribose nucleic acid (DNA) which is based on deoxyribose. A single helix has a long twisted structure, and a complementary base pair is formed inside the molecule to have a partially double helix structure. Unlike deoxyribonucleic acid, uracil is paired with adenine instead of thymine. Ribonucleic acid acts in the process of protein synthesis, and some viruses have ribonucleotides instead of deoxyribonucleic acid as a genetic material. Ribonucleic acid is composed of ribosomal ribonucleic acid (rRNA) which constitutes ribosomes according to function and structure, messenger ribonucleic acid (mRNA) which has genetic information on protein synthesis transferred from deoxyribonucleic acid, amino acid corresponding to codon of messenger ribonucleic acid Transporting ribonucleic acid (tRNA).

All organisms have several ribonucleolytic enzymes. Ribonucleic acid degrading enzyme (RNase) is an enzyme that promotes the degradation of ribonucleic acid. When the ribonucleic acid is no longer needed, the ribonucleic acid decomposing enzyme (RNase) RNA) plays an important role in the maturation process of ribonucleic acid. Ribonucleic acid degrading enzymes play an important role in various biological processes such as inhibition of neovascularization and self-incompatibility in plants. The ribonucleic acid degradation system is also the first defense mechanism against ribonucleic acid viruses and plays an important role in intracellular immune mechanisms such as ribonucleic acid interference (RNA interference). Ribonucleic acid degrading enzymes are divided into endoribonuclease and exoribonuclease according to the action mechanism. Endoribonucleic acid degrading enzyme recognizes specific nucleotide sequences of one strand of RNA and cuts out various kinds of nucleotides. RNase A, RNase H, RNase I, RNase III, RNase L, RNase P, RNase PhyM, RNase T1, RNase T2, RNase U2, RNase V1, exoribonuclease is an enzyme that removes bases from the 5 'to 3' or 3 'to 5' direction of RNA from the end, and is a polynucleotide phospholyase (PNPase), RNase PH, RNase II, RNAse R, RNase D, RNase T, oligoribonuclease, , And Exoribonuclease II.

Ribonucleotides in all cells are involved in a variety of mechanisms including 5'end capping, 3'end polyadenylation, and folding within the RNA-protein complex. Lt; RTI ID = 0.0 > ribonuclease. ≪ / RTI > Another protective mechanism is by the ribonucleic acid protease inhibitor. The ribonucleic acid protease inhibitor inhibits the activity of the ribonucleic acid degrading enzyme to prevent degradation of the ribonucleic acid.

The ribonuclease inhibitor (RNase inhibitor) is a large protein consisting of about 450 amino acid residues. It is characterized by a large amount of cysteine and leucine compared with other proteins. It is a major intracellular protein that accounts for 0.1% by weight of cellular proteins and plays an important role in regulating RNA life. The α-helix and β-sheet structures constitute ribonucleoproteinase inhibitors in a cross-over manner, and the ribonucleoproteinase inhibitors have a strong binding force to ribonucleoproteinases. The dissociation constant between them is physiological About 20 fM under the same conditions. The binding ability of ribonuclease inhibitors to ribonucleolytic enzymes plays a very important role because ribonucleic acid degrading enzymes are particularly cytotoxic to cancer cells. Studies are underway to weaken the binding ability with ribonucleoproteinase inhibitors by changing the protein structure while maintaining the activity of the ribonucleic acid degrading enzyme. Therefore, ribonucleic acid degrading enzymes have become a target of new anticancer drugs, and it is important to avoid the activity of ribonucleic acid protease inhibitors.

In addition, microRNA (miRNA), which is emerging as a new biocontrol agent in the 21st century, is very small, consisting of about 21 nucleotides, but it binds complementarily to mRNA and highly selectively regulates all gene expression in cells, It is known to play a role in all cell reactions such as differentiation, aging, and activity. Recently, it has been shown that miR-125b regulates FGRF2 to participate in psoriasis, Let-7 miRNA indirectly increases the expression of collagen type 1 in dermal fibroblasts, miR-152 and mir-181a are extracellular It has been reported that miR-434-5p is involved in the whitening of skin cells. It is known that microribonucleic acid has a new role in skin aging.

Therefore, applying micro ribonucleic acid, which can suppress skin aging such as elasticity, whitening, moisturizing, to cosmetics, can make a new category of anti-aging products.

However, ribonucleic acid degrading enzymes are present in the hands, sweat, saliva and the like of the human body. Therefore, there is a problem in the stability of the ribonucleic acid after application due to the characteristics of cosmetics applied to the skin. Therefore, there is a need for a method for inhibiting the activity of ribonucleolytic enzymes in cosmetics and enhancing the stability of ribonucleic acid.

Accordingly, the present inventors have studied activities related to ribonucleic acid for various natural products.

Accordingly, the inventors of the present invention selected ginseng, a perennial herbaceous plant of Araliaceae, among various natural products, and produced an extract from the ginseng to measure ribonuclease activity inhibitory effect. As a result, it was found that ginsenoside can be expected to have a ribonucleoproteinase inhibitor effect because of its excellent inhibitory effect on ribonucleic acid protease activity.

It is an object of the present invention to provide a cosmetic composition which contains ginseng extract to enhance the stability of ribonucleic acid useful for skin, thereby maintaining its efficacy.

It is also an object of the present invention to provide a cosmetic composition which exhibits excellent effects through preparation of supercritical, ultrasonic and fermented extracts and purification after ginseng extract to maximize its efficacy.

Accordingly, it is an object of the present invention to provide a cosmetic composition containing ribonucleic acid and ginseng extract as an active ingredient.

Preferably, the cosmetic composition is characterized by having an anti-aging effect and a whitening effect by inhibiting ribonucleic acid degrading enzyme activity.

Also preferably, the ribonucleic acid is selected from the group consisting of transporting ribonucleic acid (tRNA), messenger ribonucleic acid (mRNA), ribosomal ribonucleic acid (rRNA), microribonucleic acid (miRNA), small interfering ribonucleic acid (siRNA) non-coding RNA).

Preferably, the ginseng extract enhances the stability of the ribonucleic acid.

Preferably, the ginseng extract is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition, and the ribonucleic acid may be contained in an amount of 0.000001 to 30.0% by weight based on the total weight of the cosmetic composition.

Preferably, the ginseng extract is selected from the group consisting of (a) water, an anhydrous or a lower alcohol having 1 to 4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, chloroform, butyl acetate, diethyl ether, (B) a supercritical extraction method, (c) an ultrasonic extraction method, or (d) a fermentation extraction method, wherein the extraction is performed using an extraction solvent selected from the group consisting of methanol, hexane and mixtures thereof.

The ginseng extract according to the present invention exhibited an effect of inhibiting ribonucleolytic enzyme activity (Experimental Example 1), an effect of inhibiting microorganism activity (Experimental Example 2), an effect of inhibiting ribonucleic acid protease activity in Experimental Example 3 ), And has the effect of enhancing the whitening effect with the micro ribonucleic acid in the cosmetic formulations (Experimental Example 4).

In addition, according to the present invention, the ginseng extract extracted through ultrasound, supercritical extraction and fermentation process has further improved ribonuclease activity inhibiting effect.

FIG. 1 shows the inhibition rate of ribonucleolytic enzyme activity of ginseng extract according to the present invention.
FIG. 2 shows the inhibitory effect of the ginseng extract according to the present invention on the activity of ribonucleolytic enzymes against microRNAs.
3 shows the inhibition rate of ribonucleolytic enzyme activity of the cosmetic composition containing the ginseng extract according to the present invention.

The ginseng (reed, gold stamens, inhame, red liquor, blood ginseng, yellow ginseng, yanshan ginseng, starch ginseng), which is an effective ingredient of the cosmetic composition of the present invention, is a perennial herbaceous plant belonging to Araliaceae, and its scientific name is Panax ginseng . It is often grown as a plant growing in deep mountain regions. Its height reaches 60㎝, rhizomes are short, stand straight or diagonally, and bottom like bellflower develops. At the end of the rhizome, one main stem comes out, and at the end, 3 or 4 leaves are alive and 5 leaves are long. It is widely cultivated in Korea and China. Ginseng is used for body weakness, boredom, fatigue, loss of appetite, vomiting and diarrhea. It helps the lung function and increases the reflex activity and new function.

The pharmacological actions of ginseng include cerebral cortex excitation and inhibition, anti-fatigue, immunity enhancement, cardiac contraction, hyperglycemia inhibition, promotion of protein synthesis, homeostasis, anticancer and detoxification. Also, it has been reported that ginsenoside F5 of ginseng leaves has a whitening and anti-inflammatory effect, and that ginseng fruit inhibits melanin synthesis and has a skin whitening effect. In order to distinguish ginseng from herbs of other countries, Korean ginseng is named Korean ginseng, foreign ginseng ginseng, oriental ginseng, and ginseng ginseng. The main active ingredient of ginseng is known as a complex carbohydrate called saponin or ginsenoside, and it is known that Korean ginseng contains 37 kinds of ginsenosides Rf, Rg and Rh.

In the present invention, the ginseng extract means an extract obtained from various organs or parts (e.g., leaves, flowers, roots, stems, fruits, etc.) of ginseng, preferably an extract obtained from the seedling or root, Means an extract obtained from the outpost.

In addition, the ginseng extract of the present invention can be prepared by using a conventional solvent according to a conventional method known in the art, that is, under the conditions of ordinary temperature and pressure, but preferably (a) water, Propanol and butanol), propylene glycol, 1,3-butylene glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane (such as methanol, (B) solvent extraction using a solvent selected from the group consisting of chloroform, hexane and mixtures thereof; (b) supercritical extraction with carbon dioxide and supercritical extraction with high temperature; or (c) extraction with ultrasonic extraction.

In the present invention, a solvent extraction method using an extraction solvent is preferably used.

The ginseng extract in the present invention may be dissolved in various extraction solvents, for example, water, anhydrous or lower alcohol having 1 to 4 carbon atoms (e.g., methanol, ethanol, propanol and butanol), propylene glycol, And at least one extraction solvent selected from the group consisting of glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane and mixtures thereof. (v / v) ethanol or water, and most preferably 70% (v / v) ethanol.

It is apparent to those skilled in the art that the extract of the present invention can be obtained not only by the above-mentioned extraction solvent but also by other extraction solvent, which exhibits substantially the same effect.

In addition, the ginseng extract of the present invention includes not only the extract obtained by the above-mentioned extraction solvent, but also the extract obtained through ordinary purification and fermentation process. For example, decompression by carbon dioxide, extraction by supercritical extraction with high temperature, extraction by extraction using ultrasound, separation by ultrafiltration membrane with constant molecular weight cutoff value, various chromatography (size, charge, hydrophobic or affinity And the active fraction obtained through various purification and extraction methods, such as an extract obtained by fermentation products using natural or various microorganisms, are also included in the extract of the present invention.

Further, the present invention provides a cosmetic composition characterized in that the above extract is obtained by liquid-freezing obtained by cold-pressing and heating filtration at room temperature, and further by concentrating or lyophilizing the solvent under reduced pressure.

The supercritical fluid extraction refers to supercritical fluid extraction. Generally, the supercritical fluid is extracted from the liquid and the liquid when the gas reaches the critical point at high temperature and high pressure. (J. Chromatogr. A. 1998; 479: 200-205). In addition, it has been reported that the supercritical fluid has a polarity similar to that of a nonpolar solvent.

Carbon dioxide is a supercritical fluid with both liquid and gaseous nature, with the operation of supercritical fluid equipment reaching its critical pressure and temperature, resulting in increased solubility in fat-soluble solutes. When supercritical carbon dioxide passes through an extraction vessel containing a certain amount of sample, the lipophilic substance contained in the sample is extracted into supercritical carbon dioxide.

When the supernatant carbon dioxide containing a small amount of cosolvent is passed through the sample remaining in the extraction vessel after extracting the lipid-soluble substance, components that were not extracted by pure supercritical carbon dioxide can be extracted.

The supercritical fluid used in the supercritical extraction method of the present invention can effectively extract an active ingredient by using a mixed fluid in which a cosolvent is further mixed with supercritical carbon dioxide or carbon dioxide.

These co-solvents may be used alone or in admixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane and diethyl ether.

Most of the extracted samples contain carbon dioxide. Since the carbon dioxide is volatilized into air at room temperature, the extract obtained by the above method can be used as a cosmetic composition, and the cosolvent can be removed by a reduced pressure evaporator.

In addition, the ultrasonic extraction method is an extraction method using energy generated by ultrasonic vibration. Ultrasonic waves can destroy an insoluble solvent contained in a sample in a water-soluble solvent. Due to the high local temperature generated at this time, Since the kinetic energy of the particles is increased, sufficient energy required for the reaction is obtained. By inducing the high pressure by the shock effect of the ultrasonic energy, the mixing efficiency of the substance contained in the sample and the solvent is increased, thereby increasing the extraction efficiency.

As the extraction solvent which can be used for the ultrasonic extraction method, one or a mixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane and diethyl ether can be used. The extracted sample is recovered by vacuum filtration, and the filtrate is recovered, and the extract is removed by a vacuum evaporator and freeze-dried to obtain an extract.

The ginseng extract according to the present invention also includes a fermented extract. The fermented extract of ginseng can be prepared as follows. The ginseng is finely crushed to a size of about 100 to 500 mesh, then 1 to 50 g / L of a conventional microorganism culture solution is added, and microorganisms such as yeast strain or lactic acid bacteria are added in an amount of 10,000 to 100,000 cfu / L. The culture temperature is 30 to 37 ° C. The pH is 5 to 7 and aerobic or anaerobic conditions are maintained for about 5 to 10 days. Which can be obtained through aging and filtration.

According to the present invention, the ginseng extract is contained in an amount of 0.0001 to 30.0 wt% based on the total weight of the cosmetic composition, and more preferably, the ginseng extract is contained in an amount of 0.01 to 10 wt% based on the total weight of the cosmetic composition do. When the content of the extract is less than 0.0001% by weight, the ribonucleoproteinase activity inhibitory effect is not exhibited. When the content of the extract is more than 30.0% by weight, There is a problem with safety and stability of the product and it is not economical.

According to the present invention, the ribonucleic acid is contained in an amount of 0.000001 to 30.0% by weight, more preferably 0.00001 to 10% by weight, based on the total weight of the cosmetic composition, based on the total weight of the cosmetic composition. When the content of the extract is less than 0.000001% by weight, the anti-aging effect is not exhibited. When the content is more than 30.0% by weight, the degree of increase of the anti-aging effect against the increase of the content is insignificant, It is not economical.

In the present invention, the ribonucleic acid includes a transporting ribonucleic acid (tRNA), a messenger ribonucleic acid (mRNA), a ribosomal ribonucleic acid (rRNA), a microRNA (miRNA) and a small interfering ribonucleic acid (siRNA) Ribonucleic acid.

Therefore, the cosmetic composition of the present invention contains a ribonucleic acid having an excellent anti-aging activity and a ginseng extract having an excellent inhibitory effect on the activity of ribonuclease degradation, and has an anti-aging effect that is more improved than that of each component do.

The ribonucleic acid degrading enzyme referred to in the present invention is an endoribonucleic acid degrading enzyme or an exoribonuclease degrading enzyme.

Specifically, the endoribonucleic acid degrading enzyme includes RNase A, RNase I, RNase III, RNase H, RNase L, RNase P, RNase PhyM, RNase T1, RNase T2, RNase U2, RNase V1 and RNase V.

Specific examples of the exolibonucleolytic enzymes include polynucleotide phospholyase (PNPase), RNase PH, RNase D, RNase II, RNAse R, RNase T, oligoribonuclease, Exoribonuclease I and Exoribonuclease II.

The ingredients contained in the cosmetic composition of the present invention may contain, as an active ingredient, the ingredients commonly used in cosmetic compositions in addition to the above-mentioned effective ingredients, and may contain conventional ingredients such as antioxidants, stabilizers, solubilizers, vitamins, Supplements, and carriers.

The cosmetic composition of the present invention can be prepared into any of the formulations conventionally produced in the art and can be used in the form of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, , Oil, powdered foundation, emulsion foundation, wax foundation, pack, massage cream and spray, but is not limited thereto. More specifically, it can be manufactured in the form of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

When the formulation of the present invention is a paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

In the case where the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. Especially, in the case of a spray, a mixture of chlorofluorohydrocarbons, propane / Propane or dimethyl ether.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether Alkylamido betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

When the cosmetic composition of the present invention is a soap, a surfactant-containing cleansing formulation or a surfactant-free cleansing formulation, it may be applied to the skin and then wiped off, washed or rinsed with water. The surfactant-containing cleansing formulation may be a cleansing foam, a cleansing water, a cleansing towel and a cleansing pack. The surfactant-free cleansing formulation may be a cleansing cream, , Cleansing lotion, cleansing water and cleansing gel, but is not limited thereto.

On the other hand, the cosmetic process of the present invention refers to all cosmetic processes using the cosmetic composition of the present invention. That is, all methods known in the art using a cosmetic composition belong to the cosmetic method of the present invention.

The cosmetic composition of the present invention may be used alone or in combination, or may be used in combination with other cosmetic compositions other than the present invention.

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

Manufacturing example  1. Manufacture of ginseng extract

The shrubs of shredded ginseng were refluxed for 3 hours in 70% (v / v) aqueous ethanol solution for 5 hours, cold-frozen, and filtered through Whatman # 4 filter paper. The filtered extract was concentrated under reduced pressure and freeze-dried at 50 DEG C or lower.

Manufacturing example  2 to 3. Supercritical  Manufacture of fluid extracts and ultrasonic extracts

The supercritical extract was obtained by extracting with a conventional supercritical extraction method (extracting with ethanol as a co-solvent in a supercritical state under a pressure of about 300 atm at a temperature of about 60 DEG C). The extract was obtained by extraction using ultrasound (extraction with supersonic ultrasonic device at 30 DEG C for about 2 hours at an intensity of 25 KHz) (Production Example 3). In the case of Production Example 3, ethanol was used as a co-solvent.

Manufacturing example  4. Manufacture of ginseng fermented extract

The shredded ginseng is finely crushed to a size of about 100 to 500 mesh, then 1 to 50 g / L of a conventional microorganism culture solution is added, and microorganisms such as yeast strains or lactic acid bacteria are added in an amount of 10,000 to 100,000 cfu / L. The culture temperature is 30 to 37 ° C. The pH is 5 to 7 and aerobically or normally anaerobic conditions are incubated for about 5 to 10 days. Thereafter, the fermented extract was obtained through aging and filtration (Production Example 4).

Experimental Example  1. Inhibitory effect of ginseng extract on ribonucleic acid degrading enzyme

In Experimental Example 1, the effects of the ginseng extracts obtained in Production Examples 1 to 4 on the activity of ribonuclease degrading activity were measured.

The total ribonucleic acid used in the present invention was extracted from fibroblasts (Korean Cell Line Bank, Korea, 1x10 ⁶ cells / rxn), which is human normal skin cells, using GenElute ^™ Mammalian Total RNA Miniprep Kit (Sigma Aldrich, USA). The process of extracting whole ribonucleic acid will be briefly described as follows. First, add 250 μl of lysis buffer containing 2-mercaptoethanol to the collected fibroblasts (1 × 10 ⁶ cells), and mix well with a vortexer to disrupt the cells. Transfer to the GenElute filtration column and centrifuge for 2 minutes at 12,000 - 16,000 g. After adding 250 μl of 70% ethanol, transfer the eluate / ethanol (lysate / ethanol) to the GenElute Binding Column, mix well and centrifuge for 15 seconds at 12,000 - 16,000 g. Discard the supernatant. Add 500 μl wash solution I and centrifuge for 15 seconds at 12,000 - 16,000 g to discard the supernatant. After transferring the binding column to a new collection tube, add 500 μl of wash solution II, centrifuge at 12,000 - 16,000 g for 2 minutes and discard the supernatant. Transfer the binding column to a new collection tube, add 50 μl of elution solution, and centrifuge for 1 minute to extract the entire RNA (RNA).

The extracted ribonucleic acid was measured absorbance at 260 nm and 280 nm using a nano-drop instrument and the overall yield and purity were confirmed.

Generally, it is known that about 120 units of ribonucleolytic enzymes per ml of serum are present in humans. Pooled Human Normal Serum (Innovative Research, USA) was purchased for screening natural products with high activity inhibitory potency against various endoribonucleic acid degrading enzymes present in human serum and diluted 1/10 with sterilized PBS buffer And then used in the experiment.

20 μl of the ginseng extracts of Production Examples 1 to 4 and 50 μl of human serum were mixed with the buffer in a final volume of 0.5 ml and reacted at 22 ° C. for 15 minutes. The concentration of ginseng extract used in the test was 2,500ppm, 12,500ppm, and 25,000ppm, respectively, and then 20μl of the extract was reacted to obtain final concentrations of 100ppm, 500ppm and 1000ppm, respectively. 100 μg of human whole ribonucleic acid was added thereto and reacted at 37 ° C for 5 minutes. After completion of the reaction, 0.5 ml of 10% TCA was added to precipitate a ribonucleic acid having a large molecular weight, and the precipitated ribonucleic acid was recovered to measure the amount of ribonucleic acid at 260 nm. As a control group, 5 μl of RiboLock ™ RNase inhibitor (500 U / ml, manufactured by Thermo Scientific, USA) was used as a control, without Ganoderma lucidum extract and human serum, and human serum as a negative control. ) And human serum.

The inhibition rate (%) of ribonucleolytic enzyme activity was calculated using Equation 1, and the results are shown in FIG.

1, it was confirmed that the ginseng extract of the present invention inhibited ribonucleolytic enzyme activity in a concentration-dependent manner. Compared with the ribonucleic acid protease inhibitor (RiboLock ™), which inhibits the activity of ribonucleic acid degrading enzyme, the ginseng extract has a similar inhibitory effect on ribonucleolytic enzyme activity. On the other hand, most of the ribonucleic acid was decomposed in the negative control group which did not contain both the ginseng extract and the ribonuclease inhibitor.

Experimental Example 2. Inhibitory Effect of Ginseng Extract on Microribonucleolytic Activity

In Experimental Example 2, the effect of the ginseng extract obtained in Preparation Example 1 on the activity of ribonucleic acid degrading enzyme was measured.

The micro-ribonucleic acid used in the present invention was synthesized at 10 nmole in Genolruch (note). Pooled Human Normal Serum (Innovative Research, USA) was purchased for screening natural products with high activity inhibitory potency against various endoribonucleic acid degrading enzymes present in human serum, diluted 1/10 with sterilized PBS buffer And then used in the experiment. The synthesized micro-ribonucleic acid is known to have a whitening effect by inhibiting the activity of tyrosinase with miR-434-5p. The base sequence is 5'-GCU CGA CUC AUG GUU UGA ACC A -3 '.

In order to confirm the inhibitory effect of ginseng extract on ribonuclease activity, 10 μl of microRNA was diluted to 10 mg / ml with 100 μl of RNase free water. 10 μl of the diluted micro ribonucleic acid, 500 μg of Preparation Example 1, and 50 μl of human serum were mixed and reacted in a final volume of 500 μl at room temperature for 15 minutes. As a control, ginseng extract and human serum were not used. As a positive control, 5 μl of RiboLock ™ RNase Inhibitor (500 U / ml, Thermo Scientific, USA) was used instead of ginseng extract. After completion of the reaction, the reaction mixture was subjected to electrophoresis at 120 V for 1 hour in 2% TBE agarose gel, thereby confirming the inhibitory effect of ginseng extract on ribonuclease activity.

The inhibitory effect of ginseng extract on ribonucleic acid degrading enzyme activity on the micro-ribonucleic acid is shown in Fig.

Formulation Examples 1 to 4: Preparation of cosmetic composition containing ginseng extract

The cosmetic compositions containing the ginseng extract were prepared so as to contain the ginseng extracts of Production Examples 1 to 4, respectively. For comparison of the efficacy, instead of the ginseng extract, RNA inhibitor inhibitor (Comparative Formulation Example 1) and extract (Comparative Formulation Example 2) which does not contain any of the antioxidants or the RNAase inhibitor is shown in Table 1 below.

(Unit: wt%) division ingredient Formulation Example 1 Formulation Example 2 Formulation Example 3 Formulation Example 4 compare
Formulation Example 1 compare
Formulation Example 2 A Production Example 1 2.0 - - - - - Production Example 2 - 2.0 - - - - Production Example 3 - - 2.0 - - - Production Example 4 - - - 2.0 - - Ribonucleic acid 0.001 0.001 0.001 0.001 0.001 0.001 Human serum 0.001 0.001 0.001 0.001 0.001 0.001 Ribonucleoproteinase inhibitor - - - - 0.0002 - glycerin 5 5 5 5 5 5 EDTA-2Na 0.02 0.02 0.02 0.02 0.02 0.02 Purified water to 100 to 100 to 100 to 100 to 100 to 100 B PIGGY-60 Hydrogenated Castor Oil 0.5 0.5 0.5 0.5 0.5 0.5 Polyethylene glycol 400 3 3 3 3 3 3 Microcrystalline 0.3 0.3 0.3 0.3 0.3 0.3 Sorbitan stearate 0.5 0.5 0.5 0.5 0.5 0.5 Tocopheryl acetate 0.2 0.2 0.2 0.2 0.2 0.2 Cyclomethicone 0.3 0.3 0.3 0.3 0.3 0.3 BHT 0.05 0.05 0.05 0.05 0.05 0.05 C Incense, preservative Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount

Experimental Example  3. Inhibition of ribonucleolytic enzyme activity in the formulation of ginseng extract

The inhibitory effect of ribonuclease degrading enzyme activity on the cosmetic compositions of Formulation Examples 1, 2, 3 and 4 was measured as follows. Human serum and whole human ribonucleic acid were included in the formulations together to confirm that the ribonucleic acid was protected by the ginseng extracts of Production Examples 1 to 4. [ Formulation Examples 1 to 4 were prepared, and after 8 hours, the ribonucleic acid remaining in Formulation Examples 1, 2, 3 and 4 was separated to determine the amount of ribonucleic acid at 260 nm. In addition, the ribonucleic acid was isolated from the cosmetic composition (Comparative Formulation Example 1) containing the RNAase inhibitor instead of the ginseng extract and the cosmetic composition (Comparative Formulation Example 2) containing no ginseng extract or the RNAase inhibitor, Was measured. The experimental results are shown in Fig. In FIG. 3, it can be confirmed that the RNAs were hardly decomposed in the formulations containing the ginseng extracts of Production Examples 1 to 4.

Formulation examples 5 to 10. Preparation of cosmetic composition containing ginseng extract

The cosmetic compositions containing the ginseng extracts were prepared by respectively containing the ginseng extracts of Preparation Examples 1, 3 and 4 (Formulation Example 5-10). For comparison of the efficacy, (MiR-434-5p) and a ribonuclease inhibitor (RNase inhibitor) in which a cosmetic composition (Comparative Formulation Example 3), a cosmetic composition containing only a microbic acid (miR-434-5p) (Comparative Formulation Example 5), and a cosmetic composition (Comparative Formulation Example 6) containing neither ginseng extract nor microRNA nucleic acid (miR-434-5p) are shown in Table 2 below.

(Unit: wt%) division ingredient Formulation Example 5 Formulation Example 6 Formulation Example 7 Formulation Example 8 Formulation Example 9 Formulation Example 10 Comparative Formulation Example 3 Comparative Formulation Example 4 Comparative Formulation Example 5 Comparative Formulation Example 6 A Ginseng extract
(Production Example 1) 2.0 2.0 - - - - - - - - Ginseng extract
(Production Example 3) - - 2.0 2.0 - - - - - - Ginseng extract
(Production Example 4) - - - - 2.0 2.0 - - - - Niacinamide - - - - - - 2 - - - miR-434-5p 0.0001 - 0.0001 - 0.0001 - - 0.0001 0.0001 - 1,3-butylene glycol 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Ribonucleoproteinase inhibitor - - - - - - - - 0.0002 - EDTA-2Na 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 Purified water to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100 to 100 B Cetostearyl alcohol 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Glyceryl stearate 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Microcrystalline 0.7 0.7 0.7 0.7 0.7 0.7 0.7 0.7 0.7 0.7 Squalane 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Liquid paraffin 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Trioctanoin 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Polysorbate 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 Sorbitan stearate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Tocopheryl acetate 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Cyclomethicone 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 BHT 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 C Incense, preservative Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount Suitable amount

Experimental Example  4. Including micro ribonucleic acid and ginseng extract Cosmetics  Experiment of whitening effect of composition

The whitening effect test of the cosmetic composition containing the micro ribonucleic acid and the ginseng extract of the present invention was conducted on 20 black Korean women of 19 to 40 years old. In the measurement area, 10 of the arms were displayed in 1 cm length and 1 cm, and ultraviolet irradiation was used to cause skin pigmentation. The formulations 5 to 10 and Comparative Examples 3 to 6 of the cosmetic compositions were cleaned every morning and evening twice, respectively, and then the appropriate amount of the cosmetic compositions was continuously adjusted for 8 weeks. After 8 weeks, skin color (L value) was measured using Minolta CR 300 and the degree of change (ΔL) of skin color was calculated. The results are shown in Table 3 below.

DELTA L value Formulation Example 5 2.47 Formulation Example 6 18.9 Formulation Example 7 2.31 Formulation Example 8 1.75 Formulation Example 9 2.39 Formulation Example 10 1.81 Comparative Formulation Example 3 1.92 Comparative Formulation Example 4 0.38 Comparative Formulation Example 5 1.54 Comparative Formulation Example 6 0.11

As shown in Table 3, the degree of skin color change (DELTA L) after 8 weeks showed that the whitening effect in cosmetic formulation examples (Formulation Examples 5, 7 and 9) containing both ginseng extract and microRNA nucleic acid (Formulation Examples 6, 8, and 10) containing only the ginseng extract. This appears to be the result of the synergistic effect of microribonucleic acid miR-434-5p on the whitening effect of ginseng extract. On the other hand, the cosmetic formulation containing only micro-ribonucleic acid (Comparative Formulation Example 4) has a whitening effect, but the effect is inferior to that of a ginseng extract. However, it was confirmed that the cosmetic preparation containing the micro ribonucleic acid and the ribonuclease inhibitor together (Comparative Formulation Example 5) had excellent whitening effect. It seems that the ribonucleic acid decomposing enzyme present in the arm of the subject does not exhibit the whitening effect due to the degradation of the microRNA.

Therefore, it can be seen that the cosmetic composition containing the ginseng extract of the present invention has an effect of doubling the whitening effect of the micro-ribonucleic acid by protecting and stabilizing the micro-ribonucleic acid from the ribonucleic acid decomposing enzyme.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (7)

Ribonucleic acid and ginseng extract as an active ingredient. The cosmetic composition according to claim 1, wherein the cosmetic composition has an anti-aging effect and a whitening effect. 3. The method of claim 1 or 2, wherein the ribonucleic acid is selected from the group consisting of transporting ribonucleic acid (tRNA), messenger ribonucleic acid (mRNA), ribosomal ribonucleic acid (rRNA), microRNA (miRNA), small interfering ribonucleic acid Non-coding RNA, and non-translational ribonucleic acid (non-coding RNA). The cosmetic composition according to claim 1 or 2, wherein the ginseng extract enhances the stability of the ribonucleic acid. The cosmetic composition according to claim 1 or 2, wherein the ginseng extract is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition. The cosmetic composition according to claim 1 or 2, wherein the ribonucleic acid is contained in an amount of 0.0001 to 30.0% by weight based on the total weight of the cosmetic composition. The ginseng extract according to claim 1 or 2, wherein the ginseng extract is selected from the group consisting of (a) water, an anhydrous or a lower alcohol having 1 to 4 carbon atoms, propylene glycol, butylene glycol, glycerin, acetone, ethyl acetate, chloroform, (B) a supercritical extraction method, (c) an ultrasonic extraction method, or (d) a fermentation extraction method, which comprises extracting a cosmetic product, which is extracted by a solvent extraction method using an extraction solvent selected from the group consisting of ethyl ether, dichloromethane, Composition.

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