KR20030080429A - Microemulsion having 20-O-[α-L-arabinopyranosyl(1-〉6)-β-D-glucopyranosyl]-20(S)-protopanaxadiol by nano-emulsion technology, and skin care and pharmaceutical compositions for external applications utilizing thereof - Google Patents

Abstract

PURPOSE: A microemulsion particle containing 20-0-£α-L-arabinopyranosyl(1->6)-β-D-glucopyranosyl|-20(S)-protopanaxadiol separated from bioconversion of ginseng saponin is provided. The external preparation composition of skin using the same has an excellent effect of preventing skin aging, such as wrinkle improvement. CONSTITUTION: The microemulsion particle contains 10¬-8 to 99.9999% by weight of 20-0-£α-L-arabinopyranosyl(1->6)-β-D-glucopyranosyl|-20(S)-protopanaxadiol of the formula 1 as an effective ingredient and liposome containing lecithin, lecithin derivatives and a co-emulsifier, based on the total weight of the particle. In formula, R2 is OH and R3 is H. The microemulsion particle is prepared from acid hydrolysis, base hydrolysis or enzymolysis of ginseng extract and ginseng saponin. The cosmetic and pharmaceutical composition having excellent effects of proliferating skin cells and promoting collagen biosynthesis is obtained by formulation of the microemulsion particle.

Description

Fine emulsified particles containing 20-0- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol by nanoemulsification technology And an external composition for skin using the same {Microemulsion having 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol by nano-emulsion technology, and skin care and pharmaceutical compositions for external applications utilizing

The present invention is 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (the main metabolite of ginseng prepared by bioconversion method) S) -protopanaxadiol using a nanoemulsification technique in the fine emulsified particles and liposomes by the external skin composition for maximizing skin absorption, more specifically nanoemulsification techniques such as high pressure emulsion and solvent extraction method 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 () in fine emulsified particles and liposomes together with emulsifiers having excellent skin affinity such as lecithin. The present invention relates to a method for containing S) -protopanaxadiol, and to formulating the same, a cosmetic and pharmaceutical composition for preventing skin aging having an excellent effect on promoting skin cell proliferation and collagen biosynthesis by greatly improving transdermal absorption. In addition, in the formulation of 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol, In addition to liposomes, it may be collected and used in general emulsified particles of 500 nm or more and 1000 μm or less, dissolved in oil, or used as it is, or encapsulated. 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopa by applying microemulsion techniques including general methods of formulation techniques Means all methods for applying the efficacy of naxadiol as a cosmetic and a medicine.

Skin is the body's primary protective film that protects internal organs from changes in temperature and humidity, and from stimuli from the external environment such as ultraviolet rays and pollutants, and plays an important role in maintaining homeostasis such as body temperature control. However, excessive physical and chemical stimuli from the outside, stress and nutritional deficiency decrease the normal function of the skin and promote skin aging phenomenon such as loss of elasticity, keratinization and wrinkle formation. In order to maintain the skin, there have been efforts to effectively inhibit skin aging by maintaining skin's intrinsic function and activating skin cells by using cosmetics that have been enhanced with bioactive substances obtained from various animals, plants, and microorganisms.

However, conventional cosmetic raw materials had various problems such as insufficient efficacy or cause skin side effects.

Therefore, studies on raw materials having an anti-aging effect without causing skin side effects have been actively conducted, and among them, the interest in ginseng extract is very high, and steady research is being conducted. Looking at the research direction of ginseng extract in the meantime, ginseng extract (ginseng saponin) from ginseng extract (ginseng saponin), ginseng aglycon (aglycon) and ginseng saponin through the purification of ginseng saponin 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol was prepared and developed for separation and purification.

Ginseng saponins are glucose, rhamnose, xylose and arabinose in the alcoholic OH groups at the R1, R2 and R3 positions of the non-sugar moiety, a triterpene of the dammarane type. Sugars such as) has an ether-bonded structure, a total of 29 species have been identified in ginseng to date. Each component of the ginseng saponin was named ginsenoside as a glycoside contained in Shibata ginseng in 1964, and is an oleanine-based saponin in order of distance separated from thin layer chromatography (TLC). Ginsenoside-Ro and ginsenoside-Ra, -Rb1, -Rb2, -Rc, -Rd, -Re, -Rf, -Rg1, -Rg2, -Rg3 and -Rh, etc. were named.

This ginseng saponin was found to be different in chemical structure and pharmacological efficacy from the saponins of other plants contained in 750 kinds of plants. In particular, ginseng saponin was found to be very mild, not toxic by overdose, and have little hemolytic action.

In addition, it was reported that ginseng saponin was applied to human skin in the form of liposome, a complex with phospholipids, to revitalize aged skin, increase elasticity, increase hydration, and promote blood circulation ( Curri.SB, Gezz, Z, Longhi, MG, Castelpietra, R: Fitoterapia, 57, 217 (1986)) (Gezzi, A, Longhi, MG, Mazzoleni, R, Curri, SB: Fitoterapia, 57, 15 (1986) (Bombardelli, E. Curri, SB, Gariboldi, PL: Proc. 5th Intl. Ginseng Sym. Seoul Korea, 11 (1988))

Then, in order to apply ginseng saponin as a raw material of anti-aging products, the bioconversion 20-O- [α-L-arabinofyranosyl (1) that maintains the efficacy of ginseng saponin and increases skin permeability. -> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol on the skin was confirmed.

As a case using the ginseng extract and ginseng saponin, cosmetics (US Patents 5,565,207, 5,567,419, 5,578,312, 5,663,160, 5,626,868, 5,753,242, 5,747,300, 5,853,705, 6,027,728, 6,063,366, 6,221,569, 6,221,569, 6,221, 569, 6,221, 569, 6,221, 569) 5,587,167, 5,674,488, 5,665,393, 5,629,316, 5,776,460, 5,739,165, 5,916,555, 6,071,521, 6,083,512, 6,255,313 Many studies have been reported.

However, ginseng saponins have a structure in which alcoholic OH groups of R1, R2, and R3 are linked by ether bonds to the dammarane type, so that the hydrophilicity is large, and as the molecular weight increases, the skin permeability and absorption are low. The saponin itself has a hydrophilic property, so it could not pass through the stratum corneum, so ginseng saponin was difficult to enter the skin.

On the other hand, as research on metabolites of saponins has recently been conducted, it is suggested that the efficacy of ginseng saponins is an active body of enterobacterial metabolites in which saponins are degraded by intestinal bacteria rather than saponins themselves. Ginsenosides Rh1, Rh2 and F1, compounds K and 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucose, consisting of a sugar (glucose) attached to the aglycone It is known that pyranosyl] -20 (S) -protopanaxadiol and the like have pharmacological effects such as inhibiting cancer cell proliferation, inhibiting tumor growth, and enhancing anticancer activity of anticancer agents.

20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanax with two sugars removed by removing some of the sugars from ginseng saponin Although studies on diols are being actively conducted, there have been no studies on techniques for introducing them into the skin and formulation techniques. Also used to apply the physiological activity effect of 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol There are no limitations in the formulation technology, but general emulsified particles of 500 nm to 1000 um or less using general emulsification techniques and microemulsion particles of 500 nm or less and 30 nm or less using microemulsification technology, 20-O- [α-L-arabinofyranosyl (1 -> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol was envisaged.

Therefore, in the present invention, 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S)-purified from ginseng purified saponins by bioconversion. Protoparnaxadiol is the main active ingredient and it is contained in the fine emulsified particles and liposomes using skin-friendly emulsifier and nanoemulsification technology. It is intended to provide a composition.

1 is a skin cell organization chart showing the results of measuring collagen biosynthesis efficacy of Comparative Example 1.

2 is a skin cell organization chart showing the results of measuring collagen biosynthesis efficacy of Example 1.

The present invention is 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (the main metabolite of ginseng prepared by bioconversion method) The present invention relates to an external skin composition for maximizing skin absorption by containing S) -protopanaxadiol in fine emulsified particles and liposomes using nanoemulsification technology, and more specifically, nanoemulsification technology such as a high pressure emulsion method and a solvent extraction method. 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) in fine emulsified particles together with emulsifiers such as lecithin and excellent skin affinity. The present invention relates to a method for containing protopanaxadiol, and to formulating the same, to a significantly improved percutaneous absorption rate and to a skin anti-aging cosmetic and pharmaceutical composition having excellent skin cell proliferation and collagen biosynthesis promoting effect.

The above 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol is represented by the following formula (1).

In the above R ₁ is O-Glc ⁶ - ¹ Arap and (arabino pyrazol nosil), R ₂ is OH, R ₃ is H.

In general, hydrophobic substances are more effective than hydrophilic substances for effective skin penetration. In order to pass between intercellular lipids, including ceramides, which are distributed in the stratum corneum of the skin, hydrophobic substances are more effective in interacting with intracellular lipids than hydrophilic substances, and thus more freely pass through the outermost layer of skin. Because there is.

As shown in Formula 1, 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadai used in the present invention. Ole is characterized by a reduced molecular weight and hydrophobicity by removing some of the sugars from common ginseng saponins.

Hereinafter, the present invention will be described in more detail.

20-O-[[alpha] -L-arabinopyranosyl (1-> 6)-[beta] -D-glucopyranosyl] -20 (S) -protopanaxadiol used in the present invention is a ginseng purified saponin acid. After hydrolysis with alkali or enzyme to remove sugar from ginseng saponin, the reaction solution is prepared by passing through a silica column.

The enzymes used here include β-glucosidase, α, β-arabinosidase, and α, β-rhamnose enzymes (α, exo-glucose degrading enzymes such as β-rhamosidase), and complex enzymes containing them.

Without applying a separate nanoemulsification technology of the present invention, it is also possible to formulate by dissolving in a general oil, 20-O- [α-L- arabinofyranosyl ( 1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol is contained in an amount of 10 ^{-10 to} 70% by weight based on the total weight of the emulsified particles and liposomes, and 20-O Fine emulsified particles containing [[alpha] -L-arabinopyranosyl (1-> 6)-[beta] -D-glucopyranosyl] -20 (S) -protopanaxadiol differ depending on the preparation method, but It is contained in the amount of 10 ^{-10 to} 99.99% by weight relative to the total weight of the composition. In addition to the method of applying the technology of collecting inside the fine emulsified particles in the emulsified particles and capsules by a general emulsification method having a particle size of 500um to 1000um, or 20-O- [α-L-arabinofyranosyl 1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol can be used by dissolving in oil.

The fine emulsified particles of the present invention have a particle diameter of about 500 nm to 30 nm, preferably about 300 nm to 50 nm. In the case of a conventional emulsified particles, the microemulsion particles and liposomes of the present invention have a size of at least 500 nm or more, and thus the percutaneous absorption available area also increases due to a relatively increased contact area with the skin. In addition, in consideration of the fact that the intercellular lipids in the stratum corneum layer is about 50 nm and the emulsification film of the emulsified particles is flexible, the fine emulsified particles of the present invention are easily absorbed and diffused into the intercellular lipids. It was.

That is, emulsified particles having an average particle diameter of 300 nm to 50 nm prepared by nanoemulsification technology are obtained through the two paths of increasing the contact area with the skin and penetration and diffusion into intercellular lipids. Percutaneous absorption of 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol is enhanced.

On the other hand, lecithin, which is an emulsifier used in the present invention, contains 0.5 to 5% by weight, preferably 2 to 3% by weight of the total weight of the emulsified particles, and the components of the lecithin include phosphatidylcholine, lysophosphatidylcholine, Unsaturated choline-based, serine-based, ethanolamine-based compounds such as phosphatidylethanolamine, and hydrogenated forms thereof.

In addition, the co-emulsifier used in conjunction with the lecithin in the present invention may be an anionic, cationic, nonionic or zwitterionic emulsifier, 0.5 to 5 times the lecithin content, depending on the content and composition of the lecithin used, Preferably it is used in the ratio of 1-3 times.

In addition, 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl was used as a nanoemulsification technique using a high pressure emulsification method (pressure of 500-2,500 bar) and a solvent extraction method. ] -20 (S)-Protopananaxadiol containing fine emulsified particles.

Skin anti-aging cosmetics containing 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol according to the present invention And the pharmaceutical composition is not particularly limited in its formulation. For example, supple cosmetics, astringent cosmetics, nourishing cosmetics, nourishing cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, body lotion, body cream, body oil and body It can be formulated into cosmetics such as essences, makeup bases, foundations, hair dyes, shampoos, rinses, body cleansers, toothpastes, mouthwashes, and lotions, ointments, gels, creams, patches or sprays.

Hereinafter, the configuration and effects of the present invention will be described in more detail with reference to Examples and Test Examples.

Reference Example 1 Preparation of Ginseng Purified Saponin

4 g of ethanol including water and water was added to 2 kg of red ginseng, and the mixture was extracted under reflux three times at 77 ° C. and then deposited at 15 ° C. for 6 days. Thereafter, the residue and the filtrate were separated through filter cloth filtration and centrifugation. The extract obtained by concentrating the separated filtrate under reduced pressure was suspended in water, and then extracted five times with 1 L of ether to remove the pigment, and the aqueous layer was 1-butanol. Extracted three times with 500 ml. The total 1-butanol layer obtained therefrom was treated with 5% KOH, washed with distilled water, and then concentrated under reduced pressure to obtain 1-butanol extract, which was dissolved in a small amount of methanol and added to a large amount of ethyl acetate. By drying the precipitate, 100 g (yield: 5%) of ginseng purified saponin was obtained, and a total of 1 kg of ginseng purified saponin was prepared by repeating the same operation 10 times.

REFERENCE EXAMPLE 2 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol by acid hydrolysis method Manufacture

20 g (v / w) of 7% sulfuric acid / 50% ethanol solution (v / w) was added to 100 g of purified ginseng saponin obtained in Reference Example 1, and the mixture was heated and refluxed in a 100 ° C. water bath for 6 hours to bind to ginseng saponin. Sugar bonds were hydrolyzed. The reaction solution was concentrated under reduced pressure, the solvent was removed, distilled water (1,000 ml) was added to the residue and suspended, and then extracted three times with the same amount of ether. The total ether layer was washed with distilled water and then dehydrated with anhydrous magnesium sulfate (MgSO ₄ ), filtered and concentrated to obtain a crude product. The obtained crude product was separated by silica gel column chromatography (chloroform: methanol = 9: 1-> 4: 1, gradually increasing polarity), and thin layer chromatography (chloroform / methanol / water = 65) for each fraction was performed. / 35/10, Rf = 0.49) using 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadai The fractions of ol are isolated and recovered to give 700 mg of 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol 700 mg (Yield 0.7%) was obtained.

REFERENCE EXAMPLE 3 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol by base hydrolysis method Manufacture

100 g of ginseng purified saponin obtained in Reference Example 1 was dissolved in dry pyridine (5000 ml), and sodium methoxide (powder, 100 g) was added thereto to reflux for 8 hours on an oil bath to hydrolyze the sugar bond of saponin. The reaction solution was concentrated under reduced pressure, the solvent was removed, distilled water (1,000 ml) was added to the residue and suspended, and then extracted three times with the same amount of ether. The total ether layer was washed with distilled water and then dehydrated with anhydrous magnesium sulfate (MgSO ₄ ), filtered and concentrated to obtain a crude product. The obtained crude product was separated by silica gel column chromatography (chloroform: methanol = 9: 1-> 4: 1, gradually increasing polarity), and thin layer chromatography (chloroform / methanol / water = 65) for each fraction was performed. / 35/10, Rf = 0.49) using 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadai The fractions of ol are separated and recovered to give 850 mg of 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol (Yield 0.85%) was obtained.

[Reference Example 4-1] 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadai through enzymatic digestion Manufacturing

100 g of ginseng purified saponin obtained in Reference Example 1 was dissolved in 1000 ml of citrate buffer (pH 5.5), and 1 g of naringinase enzyme isolated in penicillium was added and stirred for 48 hours in a 40 ° C water bath. The reaction was carried out. After periodic confirmation by thin layer chromatography, when the substrate was completely lost, the reaction was terminated by heating in hot water for 10 minutes, and then the reaction solution was extracted and concentrated three times with the same amount of ether. The obtained product was separated by silica gel column chromatography (chloroform: methanol = 9: 1-> 4: 1, gradually increasing polarity), and thin layer chromatography (chloroform / methanol / water = 65 / 35/10, Rf = 0.39) using 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol Fractions were separated and recovered and finally 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol 1400 mg ( Yield 1.4%).

Reference Example 4-2

100 g of ginseng purified saponin was dissolved in 1000 ml of citrate buffer solution (pH 4.0) containing 15% ethanol, and 0.5 g of naringinase enzyme isolated in penicillium was added thereto for 48 hours in a 40 ° C water bath. The reaction was carried out while stirring. After periodic confirmation by thin layer chromatography, when the substrate was completely lost, the reaction was terminated by heating in hot water for 10 minutes, and then the reaction solution was extracted and concentrated three times with the same amount of ethyl acetate. The obtained product was separated by silica gel column chromatography (chloroform: methanol = 9: 1-> 4: 1, gradually increasing polarity), and thin layer chromatography (chloroform / methanol / water = 65 / 35/10, Rf = 0.39) using 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol Fractions were separated and recovered, and finally 900 mg of 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol Yield 0.9%).

Reference Example 4-3

100 g of ginseng purified saponin was dissolved in 1000 ml of citrate buffer solution (pH 4.0) containing 15% ethanol, and 2 g of pectinase enzyme isolated from Aspergillus was added thereto and stirred for 48 hours in a 30 ° C water bath. The reaction was carried out. After periodic confirmation by thin layer chromatography, when the substrate was completely lost, the reaction was terminated by heating in hot water for 10 minutes, and then the reaction solution was extracted and concentrated three times with the same amount of ethyl acetate. The obtained product was separated by silica gel column chromatography ((chloroform: methanol = 9: 1-> 4: 1 with increasing polarity) and separated by thin layer chromatography (chloroform / methanol / water = 65). / 35/10, Rf = 0.49) using 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadai The fractions of ol are isolated and recovered to give 1100 mg of 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol (Yield 1.1%) was obtained.

Reference Example 4-4

100 g of ginseng purified saponin was dissolved in 1000 ml of citrate buffer (pH 5.5), and 2 g of pectinase enzyme isolated in Aspergillus was added thereto and reacted with stirring for 30 hours in a 30 ° C water bath. After periodic confirmation by thin layer chromatography, when the substrate was completely lost, the reaction was terminated by heating in hot water for 10 minutes, and then the reaction solution was extracted and concentrated three times with the same amount of ether. The obtained product was separated by silica gel column chromatography (chloroform: methanol = 9: 1-> 4: 1, gradually increasing polarity), and thin layer chromatography (chloroform / methanol / water = 65 / 35/10, Rf = 0.39) using 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol Fractions were separated and recovered, and finally 1200 mg of 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol ( Yield 1.2%).

Nano-emulsified particles containing 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol obtained from the above reference example Was prepared as in Examples 1 to 3.

Example 1

20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopa dissolved in lecithin, hydrogenated lecithin, cholesterol, soybean oil and propylene glycol Completely dissolve Naxadiol to 70 ~ 75 ℃, mix with pre-heated water-part (distilled water, EDTA), pre-emulsion with regular homomixer (3,000 ~ 6,000rpm for 3 minutes). ) The test was performed at 1,000Bar / 3cycles using a microfluidizer. In the case of hydrogenated lecithin, the emulsion has excellent stability, but in terms of skin absorption, the skin affinity is inferior to that of unsaturated lecithin. Thus, two lecithins were mixed and used.

Example 2

20-O- [α-L-arabinofyranosyl (1-) dissolved in lecithin, PEG-5 rapeseed sterols, capric / caprylic triglycerides, BHT, a-tocopherol and pentylene glycol and ethanol > 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol, dissolved completely by heating to 70-75 ° C, then mixed with pre-heated water-part (distilled water, EDTA) Pre-emulsification with a mixer (3,000-6,000 rpm for 3 minutes) was performed at 1,000 Bar / 3cycles using a high pressure emulsifier. Here, BHT was added as an antioxidant to compensate for the chemical instability of unsaturated lecithin, and PEG-5 rapeseed sterol was added as an auxiliary emulsifier to increase the emulsion stability.

Example 3

Hydrogenated Lecithin, Hydrogenated Lysophosphatidylcholine (HLPC), and 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) dissolved in propylene glycol and ethanol Dissolve completely by heating protopanaxadiol to 70 ~ 75 ℃, mix with pre-heated water-part (distilled water, EDTA, glycerin, betaine) and emulsify with general homomixer (3,000 ~ 6,000rpm) ) Cooled to room temperature. For reference, among the components, hydrogenated lysophosphatidylcholine is a component produced by hydrolysis of hydrogenated phosphatidylcholine (HPC) among the components constituting hydrogenated lecithin, and has better emulsifying power than HPC.

20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadai prepared in Examples 1 to 3 above In order to prepare micro-emulsion particles and liposomes and general emulsions containing ol, and to compare the percutaneous absorption difference, Comparative Examples 1 to 3 containing ginseng purified saponin were prepared. Same as 1. In Table 1 below, Example 1, Example 2, and Comparative Example 1 and Comparative Example 2 used a high-pressure emulsification method to prepare microemulsified particles, and Example 3 and Comparative Example 3 were prepared using a general homo mixer. Emulsified particles were allowed to form.

ingredient Example Comparative example One 2 3 One 2 3 Hydrogenated lecithin 1.5 - 2.5 1.5 - 2.5 lecithin 3.0 2.0 - 3.0 2.0 - PEG-5 Rape Seed Sterols - 4.0 - - 4.0 - Ceteares-10 - - - - - - Capric / Caprylic Triglycerides - 7.5 - - 7.5 - Hydrolyzed lysophosphatidylcholine - - 0.15 - - 0.15 cholesterol 1.5 - - 1.5 - - Soybean oil 7.5 - - 7.5 - - Pentylene glycol - 5.0 - - 5.0 - Propylene glycol 5.0 - 4.0 5.0 - 4.0 ethanol - 7.5 6.5 - 7.5 6.5 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol 1.5 1.5 1.5 - - - Ginseng Purified Saponin - - - 1.5 1.5 1.5 Alpha Tocopherol - 0.2 - - 0.2 - Butylhydroxytoluene - 0.01 - - 0.01 - EDTA 0.05 0.05 0.05 0.05 0.05 0.05 glycerin - - 4.0 - - 4.0 Betaine - - 1.0 - - 1.0 Distilled water to 100 to 100 to 100 to 100 to 100 to 100

Hereinafter, fine emulsified particles and liposomes containing 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol or Referring to the skin cosmetic composition formulated with the general emulsified particles are as follows.

[Prescription Example 1] Cream Preparation

Furtherance Formulation Example 1 Formulation Example 2 Formulation Example 3 Comparative Formulation Example 1 Comparative Formulation Example 2 Comparative Formulation Example 3 Example 1 10.0 - - - - - Example 2 - 10.0 - - - - Example 3 - - 10.0 - - - Comparative Example 1 - - - 10.0 - - Comparative Example 2 - - - - 10.0 - Comparative Example 3 - - - - - 10.0 Beeswax 10.0 10.0 10.0 10.0 10.0 10.0 Polysorbate 60 1.5 1.5 1.5 1.5 1.5 1.5 Solbitan Sesquioleate 0.5 0.5 0.5 0.5 0.5 0.5 PEG60 Cured Castor Oil 2.0 2.0 2.0 2.0 2.0 2.0 Liquid paraffin 10.0 10.0 10.0 10.0 10.0 10.0 Squalane 5.0 5.0 5.0 5.0 5.0 5.0 Caprylic / Caprol Triglycerides 5.0 5.0 5.0 5.0 5.0 5.0 glycerin 5.0 5.0 5.0 5.0 5.0 5.0 Butylene glycol 3.0 3.0 3.0 3.0 3.0 3.0 Propylene glycol 3.0 3.0 3.0 3.0 3.0 3.0 Triethanolamine 0.2 0.2 0.2 0.2 0.2 0.2 antiseptic Quantity Quantity Quantity Quantity Quantity Quantity Pigment Quantity Quantity Quantity Quantity Quantity Quantity Spices Quantity Quantity Quantity Quantity Quantity Quantity Purified water to 100 to 100 to 100 to 100 to 100 to 100

[Prescription 2] Nutritional Cosmetics

Furtherance Formulation Example 4 Formulation Example 5 Formulation Example 6 Comparative Formulation Example 4 Comparative Formulation Example 5 Comparative Formulation Example 6 Example 1 10.0 - - - - - Example 2 - 10.0 - - - - Example 3 - - 10.0 - - - Comparative Example 1 - - - 10.0 - - Comparative Example 2 - - - - 10.0 - Comparative Example 3 - - - - - 10.0 Cetylethylhexanoate 5.0 5.0 5.0 5.0 5.0 5.0 Cetostearyl alcohol 1.0 1.0 1.0 1.0 1.0 1.0 Lipophilic monostearic acid stearate 0.8 0.8 0.8 0.8 0.8 0.8 Squalane 2.0 2.0 2.0 2.0 2.0 2.0 Polysorbate 60 1.5 1.5 1.5 1.5 1.5 1.5 Solbitan Sesquioleate 0.5 0.5 0.5 0.5 0.5 0.5 glycerin 5.0 5.0 5.0 5.0 5.0 5.0 Triethanolamine 0.2 0.2 0.2 0.2 0.2 0.2 Carboxy Vinyl Polymer 0.2 0.2 0.2 0.2 0.2 0.2 antiseptic a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Pigment a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Spices a very small amount a very small amount AL amount a very small amount a very small amount a very small amount Purified water to 100 to 100 to 100 to 100 to 100 to 100

[Prescription 3] Flexible Cosmetics

Furtherance Formulation Example 7 Formulation Example 8 Formulation Example 9 Comparative Formulation Example 7 Comparative Formulation Example 8 Comparative Formulation Example 9 Example 1 10.0 - - - - - Example 2 - 10.0 - - - - Example 3 - - 10.0 - - - Comparative Example 1 - - - 10.0 - - Comparative Example 2 - - - - 10.0 - Comparative Example 3 - - - - - 10.0 Betaine 3.0 3.0 3.0 3.0 3.0 3.0 Natto gum 3.0 3.0 3.0 3.0 3.0 3.0 Cellulose gum 0.08 0.08 0.08 0.08 0.08 0.08 ethanol 5.0 5.0 5.0 5.0 5.0 5.0 Polyoxyethylene Cured Castor Oil 0.5 0.5 0.5 0.5 0.5 0.5 Tocopherol Acetate 0.2 0.2 0.2 0.2 0.2 0.2 antiseptic a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Pigment a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Purified water to 100 to 100 to 100 to 100 to 100 to 100

[Prescription Example 4] Gel

Furtherance Formulation Example 10 Formulation Example 11 Formulation Example 12 Comparative Formulation Example 10 Comparative Formulation Example 11 Comparative Formulation Example 12 Example 1 10.0 - - - - - Example 2 - 10.0 - - - - Example 3 - - 10.0 - - - Comparative Example 1 - - - 10.0 - - Comparative Example 2 - - - - 10.0 - Comparative Example 3 - - - - - 10.0 Disodium Ethylenediaminetetraacetate 0.02 0.02 0.02 0.02 0.02 0.02 Ethoxyglycol 1.00 1.00 1.00 1.00 1.00 1.00 Polyacrylate 20.00 20.00 20.00 20.00 20.00 20.00 ethanol 30.00 30.00 30.00 30.00 30.00 30.00 Hydrogenated castor oil 0.80 0.80 0.80 0.80 0.80 0.80 Phenyltrimethicone 0.20 0.20 0.20 0.20 0.20 0.20 Triethanolamine 0.40 0.40 0.40 0.40 0.40 0.40 incense a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Purified water to 100 to 100 to 100 to 100 to 100 to 100

[Prescription Example 5] Ointment

Furtherance Formulation Example 13 Formulation Example 14 Formulation Example 15 Comparative Formulation Example 13 Comparative Formulation Example 14 Comparative Formulation Example 15 Example 1 10.0 - - - - - Example 2 - 10.0 - - - - Example 3 - - 10.0 - - - Comparative Example 1 - - - 10.0 - - Comparative Example 2 - - - - 10.0 - Comparative Example 3 - - - - - 10.0 Caprine / Capryltriglycide 10.0 10.0 10.0 10.0 10.0 10.0 Liquid paraffin 10.0 10.0 10.0 10.0 10.0 10.0 Solbitan Sesquioleate 6.0 6.0 6.0 6.0 6.0 6.0 Octyldodeceth-25 9.0 9.0 9.0 9.0 9.0 9.0 Cetylethylhexanoate 10.0 10.0 10.0 10.0 10.0 10.0 Squalane 1.0 1.0 1.0 1.0 1.0 1.0 Salicylic acid 1.0 1.0 1.0 1.0 1.0 1.0 glycerin 15.0 15.0 15.0 15.0 15.0 15.0 Sorbitol 10.0 10.0 10.0 10.0 10.0 10.0 Distilled water to 100 to 100 to 100 to 100 to 100 to 100

[Prescription Example 6] Spray

Furtherance Formulation Example 16 Formulation Example 17 Formulation Example 18 Comparative Formulation Example 16 Comparative Formulation Example 17 Comparative Formulation Example 18 Example 1 10 - - - - - Example 2 - 10 - - - - Example 3 - - 10 - - - Comparative Example 1 - - - 10 - - Comparative Example 2 - - - - 10 - Comparative Example 3 - - - - - 10 Triethanolamine 0.2 0.2 0.2 0.2 0.2 0.2 Polyvinylpyrrolidone / Vinyl Acetate 3.0 3.0 3.0 3.0 3.0 3.0 glycerin 5.0 5.0 5.0 5.0 5.0 5.0 Polyacrylate 0.2 0.2 0.2 0.2 0.2 0.2 Distilled water to 100 to 100 to 100 to 100 to 100 to 100

The prescription is used in a pump container.

[Example 7] Patch

Furtherance Formulation Example 19 Formulation Example 20 Formulation Example 21 Comparative Formulation Example 19 Comparative Formulation Example 20 Comparative Formulation Example 21 Example 1 10 - - - - - Example 2 - 10 - - - - Example 3 - - 10 - - - Comparative Example 1 - - - 10 - - Comparative Example 2 - - - - 10 - Comparative Example 3 - - - - - 10 Polyvinyl alcohol 2.0 2.0 2.0 2.0 2.0 2.0 Polyvinylpyrrolidone 3.0 3.0 3.0 3.0 3.0 3.0 Sodium polyacrylic acid 3.0 3.0 3.0 3.0 3.0 3.0 Sodium alginate 1.0 1.0 1.0 1.0 1.0 1.0 Retinyl palmitate 1.0 1.0 1.0 1.0 1.0 1.0 Butylene glycol 3.0 3.0 3.0 3.0 3.0 3.0 Chondroitin Sulfate 1.0 1.0 1.0 1.0 1.0 1.0 Skirt Mushroom Extract 1.0 1.0 1.0 1.0 1.0 1.0 Meadowfoam oil 1.0 1.0 1.0 1.0 1.0 1.0 PEG (20) sorbitan stearate 1.0 1.0 1.0 1.0 1.0 1.0 BHT a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Zinc oxide a very small amount a very small amount a very small amount a very small amount a very small amount a very small amount Distilled water to 100 to 100 to 100 to 100 to 100 to 100

Test Example 1 Percutaneous Absorption Measurement Test

Skin absorption was measured by using Franz permeation cells on guinea pig skin. Immediately before the test, abdominal skin of the guinea pigs was taken out, cut into square 1 cm 2 areas, and then mounted in a transmission cell having a diameter of 0.9 cm in the diameter of the penis and fixed with a clamp. One side of the skin (donor container) is diluted with distilled water to 0.05 ml of Examples 1 to 3 and Comparative Examples 1 to 3 described above, and in the case of Formulation Examples 1 to 15 and Comparative Formulation Examples 1 to 15, 0.5 ml was added. The other side (receptor vessel) was to contact the solvent mixed with purified water and ethanol in a 4: 1 weight ratio, the temperature was maintained at 32 ℃ the actual skin temperature during the test. After the start of the test, a portion of the solvent was taken at regular intervals, and then 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyira was absorbed into the skin using HPLC. Nosyl] -20 (S) -protopanaxadiol was measured and expressed as the skin absorption (μg / cm 2 / wt%) per application concentration, and the results are shown in Table 2 below. For ginseng purified saponins, the content of transdermal absorbed saponins was quantified. In the case of pananaxadiol, the percutaneous absorption of 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol The total transdermal absorption was calculated by quantitative analysis of the content.

<HPLC Analysis Conditions>

-Column: C18 (ODS)

Solvent Flow: 1ml / min

-Detection UV: 203nm

Sample test concentration: 5mg / ml

-Sample injection amount: 10㎍

Eluent: Gradient condition

A: Acetonitrile / D.I. water = 15/85

B: Acetonitrile / D.I. water = 80/20

<Solvent Gradient Conditions>

Minutes A (%) B (%) 0 100 10 70 30 25 50 50 40 100 70 100

Percutaneous absorption according to elapsed time (Examples 1-3 and Comparative Examples 1-3) Example Elapsed time (hr) Comparative example Elapsed time (hr) 0 4 8 12 0 4 8 12 One 0 15.15 28.98 33.13 One 0 3.52 8.11 14.41 2 0 16.02 26.14 34.21 2 0 3.66 8.35 14.06 3 0 14.59 20.25 25.32 3 0 3.35 5.04 10.95

Percutaneous absorption according to elapsed time (Formulation Examples 1-15 and Comparative Formulation Examples 1-15) Formulation example Elapsed time (hr) Comparative formulation example Elapsed time (hr) 0 4 8 12 0 4 8 12 One 0 12.12 24.00 35.21 One 0 2.51 3.98 4.68 2 0 11.25 21.63 33.74 2 0 1.15 2.62 3.86 3 0 2.21 3.86 5.40 3 0 0.48 1.01 2.03 4 0 15.98 31.86 31.97 4 0 2.62 3.21 5.93 5 0 11.45 27.63 32.71 5 0 2.24 3.53 5.92 6 0 2.24 3.75 5.11 6 0 0.45 0.97 1.97 7 0 14.30 28.59 33.99 7 0 3.23 4.84 6.83 8 0 11.22 21.45 32.55 8 0 1.56 3.47 5.31 9 0 2.23 3.65 5.06 9 0 0.50 1.03 2.11 10 0 15.21 31.25 31.21 10 0 2.33 5.13 7.02 11 0 11.52 21.88 35.94 11 0 1.56 2.73 4.32 12 0 2.12 3.36 5.35 12 0 0.49 1.11 2.23 13 0 12.13 30.99 31.85 13 0 2.45 4.10 6.02 14 0 11.73 21.97 35.43 14 0 1.12 3.54 6.33 15 0 2.23 3.45 5.61 15 0 0.48 0.96 2.06

From the above test results, it can be seen that Examples 1 and 2, which are the case of emulsified particles prepared by applying the nanoemulsification technique, showed better transdermal absorption than those of Example 3 forming general emulsified particles. Compared to Comparative Examples 1 to 2, which are microemulsion particles prepared by using nanoemulsification technology, it was confirmed that the percutaneous absorption rate was much higher. In particular, when the nanoemulsification technique is applied to 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol ( In Examples 1 to 2, it was confirmed that there was a dramatic increase in transdermal absorption.

Furthermore, through the corresponding examples and comparative examples, 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S)- Protoparnaxadiol showed higher transdermal absorption than other glucosyl ginsenosides of ginseng saponin, which is 20-O- [α-L-arabinofyranosyl (1-> 6) -β. -D-glucopyranosyl] -20 (S) -protopanaxadiol may be derived from the characteristics of the chemical structure.

That is, 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol is much higher transdermal than ginseng purified saponin. Absorption rate, especially by applying lecithin and nanoemulsification technology excellent skin affinity was found to exhibit excellent transdermal absorption rate.

In addition, this can be confirmed through Formulation Examples 1 to 15 and Comparative Formulation Examples 1 to 15, which formulate examples and comparative examples, and 20-O- [α due to the use of lecithin having excellent nanoemulsification technology and skin affinity. It was confirmed that the transdermal absorption rate of -L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol was reproduced and applied as it is in the formulation.

Considering that the makeup duration is generally 4 to 8 hours, 20-O- [α-L-arabinofyranosyl (1-> 6) -β- compared to the examples and formulations containing ginseng purified saponin D-glucopyranosyl] -20 (S) -protopanaxadiol in Examples and Formulations can be seen that the percutaneous absorption is significantly superior.

Test Example 2 Measurement of Proliferative Effect of Fibroblasts

Human fibroblasts cultured in DMEM (Doubecco's Modified Eagle's Media) medium containing 3.5% fetal calf serum were dispensed at a 96-well microtiter plate at 5,000 cells / well, and Example 1 was used as a sample. 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol and ginseng purified saponin of Comparative Example 1, respectively It was used in an amount of 1%, and using Formulation Example 1 and Comparative Formulation Example 1 in an amount of 10% each, sequentially diluted 1/10 by a culture medium, and then incubated at 37 ° C for 4 days. After incubation, 50 μl of 0.2% MTT (3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide) solution was added to each well, followed by incubation for 4 hours at 37 ° C. Formazan was dissolved in dimethyl sulfoxide (DMSO). The absorbance of the dissolved formazan was measured at 570 nm using a microplate reader. This was carried out in the same manner as above for the control group not treated with ginseng purified saponin and biozine to measure the absorbance. Test group containing purified ginseng saponin and 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol and After comparing the absorbance of the control group without the content, the results are shown in Table 3.

Sample concentration (%) Fibroblast proliferation (%) Example 1 Comparative Example 1 Formulation Example 1 Comparative Formulation Example 1 1 × 10 ^-8 5 3 5 3 1 × 10 ^-7 16 5 12 6 1 × 10 ^-6 28 8 23 10 1 × 10 ^-5 45 13 44 13 1 × 10 ^-4 54 19 52 17 1 × 10 ^-3 66 25 64 23 1 × 10 ^-2 81 41 78 39 1 × 10 ^-1 98 47 95 43

From Table 3, 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S)-compared to fibroblasts treated with ginseng purified saponin It was found that the proliferative effect of the fibroblasts treated with the protopanaxadiol was much higher. In addition, in the case of Formulation Example 1 and Comparative Formulation Example 1 formulated in each example, it was confirmed that Formulation Example 1 has a superior fibroblast proliferation effect than Comparative Formulation Example 1.

Test Example 3 Measurement of proliferation effect of keratinocytes

Using the keratinocytes to measure the proliferation effect of the keratinocytes in Example 2, Comparative Example 2 and Formulation Example 2 and Comparative Formulation Example 2 in the same manner as in Test Example 2, the results are shown in Table 4. .

Sample concentration (%) Keratinocyte proliferation (%) Example 2 Comparative Example 2 Formulation Example 2 Comparative Formulation Example 2 1 × 10 ^-8 5 4 5 4 1 × 10 ^-7 13 6 13 6 1 × 10 ^-6 18 7 18 7 1 × 10 ^-5 25 11 25 11 1 × 10 ^-4 34 14 34 14 1 × 10 ^-3 39 19 38 18 1 × 10 ^-2 45 23 44 21 1 × 10 ^-1 53 27 51 25

As can be seen in Table 4, compared with keratinocytes treated with ginseng purified saponin, 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] It was confirmed that keratinocytes treated with -20 (S) -protopanaxadiol showed about 2 times improved proliferative effect. These results were confirmed to show the same tendency in the formulation prepared using the nanoemulsification technology and skin-friendly lecithin and the formulation containing the same.

Test Example 4 In vitro Collagen Biosynthesis Efficacy Measurement

After culturing the human fibroblasts in a 24-hole plate incubator, and diluting them sequentially with culture medium for Example 3 and Comparative Example 1 by using the same method as Test Example 2, and added to each one by one, and Formulation Example 3, comparison For Formulation Example 3 was added by diluting sequentially in culture medium 1/10. On the third day of culture, 0.5 ml of DMEM medium containing 10% fetal calf serum was added to each well, followed by 10 [mu] g of L [2, 3, 4, 5-3H] -proline. After 24 hours, the media and cells in each well were scraped, washed in 5% Trichloroacetic acid (TCA) solution, and dispensed into two test tubes, and in one test tube, Type I cola. 1 unit / μl of genase (type I collagenase) was added and incubated at 37 ° C. for 90 minutes, and other test tubes were stored at 4 ° C. Thereafter, 0.05 ml of 50% TCA was added to all test tubes and allowed to stand at 4 ° C. for 20 minutes, followed by centrifugation at 12,000 rpm for 10 minutes, respectively, and the supernatant and the precipitates were respectively liquid scintillation counter (LSC). To obtain a decay per minute (DPM) value, to obtain a collagen biosynthesis value (RCB; Relative Collagen Biosynthesis) for the control group and the test group based on Equation 1 below, the results are shown in Table 5 below .

Sample concentration (%) Collagen Biosynthesis Proliferative Capacity (%) Example 3 Comparative Example 3 Formulation Example 8 Comparative Formulation Example 8 1 × 10 ^-8 5 2 2 2 1 × 10 ^-7 7 2 5 2 1 × 10 ^-6 11 4 9 4 1 × 10 ^-5 16 6 13 6 1 × 10 ^-4 21 10 17 9 1 × 10 ^-3 29 12 24 11 1 × 10 ^-2 38 16 31 15 1 × 10 ^-1 44 20 36 19

From the results in Table 5, 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) compared to fibroblasts treated with ginseng purified saponin The fibroblasts treated with) -protopanaxadiol showed a three-fold improvement in collagen biosynthesis-promoting effects. These results showed that 20-O- [α-L-Ara was used even without nanoemulsification. The same tendency was observed in the examples prepared using Vinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol and skin affinity lecithin and formulations containing the same. Could be confirmed.

[Test Example 5] Determination of collagen biosynthesis efficacy in vivo

Hairless mouse (42 weeks, female) in 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadai All (excipient; EtOH: PG = 7: 3) was apply | coated and patched for 3 days, and it rested for 24 hours, and then repeatedly patched for 3 days. After that, the skin of the bald mice was biopsyed and tissue stained. Tissue staining was performed using type I pN procollagen, MMP-1 (Immunostaining against Matrix Metalloproteinase-1) and Haematoxylin and Eosin Staining. The changes in the expression quantity qualitative (quantitative) and skin (epidermal) thickness were observed, and the results are shown in FIGS. 1 and 2.

1 and 2, it was confirmed that the collagen biosynthesis was promoted in Example 1 rather than Comparative Example 1, which is 20-O- [α-L- arabinofyranosyl ( 1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol can be effectively delivered into the skin.

As a result, the ginseng tablet saponin is not effectively absorbed by the skin, and as a result, it was confirmed that the collagen biosynthesis effect is minimal in Examples and Formulation Examples.

This is a result showing that ginseng purified saponin is structurally difficult to absorb the skin, which is not easily overcome by the use of skin-friendly lecithin or formulation using nanoemulsification technology.

That is, from the above test results, 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S)-having a structure that facilitates transdermal absorption. By using skin-friendly lecithin and nanoemulsification technology in the transdermal absorption of ProtoPanaxadiol, it was confirmed that maximized transdermal absorption is possible, and as a result, effective collagen biosynthesis can be achieved.

Test Example 6 Skin Wrinkle Improvement Effects on Human Skin

Thirty-five subjects with facial wrinkles aged 35 to 45 years were given the cream formulation of Formulation Example 1 and Comparative Formulation Example 1, and the skin wrinkle improvement effect was evaluated. The cream of Formulation Example 1 was used on the left side of the subject and the cream of Comparative Formulation Example 1 was used on the right side for 3 months, and the skin condition was measured on both sides of the face before the use of the cream. The change in skin wrinkles was measured by the method. Skin measurement was performed in a constant temperature room with a temperature of 24 ° C. and a relative humidity of 40%. The wrinkles at the tail of the eyes were replicated, and skin wrinkles were measured using a Visiometer system (C + K). The amount of change in skin wrinkles was calculated according to Equation 2 below.

(In the above formula, Tdi; measured value at D ₉₀ , Tdo; measured value at D _0. )

According to the above formula, the calculated results are shown in Table 6 below.

Wrinkle Reduction (△%) Formulation Example 1 56 ± 15% Comparative Formulation Example 1 21 ± 10%

As can be seen from the above results, in the present invention, in order to overcome the disadvantage of low transdermal absorption due to the structural characteristics of ginseng saponin, 20-O- [α-L-arabinofyranosyl (1-> 6) from which sugar was removed -(beta) -D-glucopyranosyl] -20 (S) -protopanaxadiol is contained inside fine emulsion particles using skin-friendly emulsifier and nanoemulsification technology to maximize transdermal absorption and reduce skin aging such as wrinkle reduction. The prevention effect was found to be excellent.

Claims (12)

20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) represented by Chemical Formula 1 prepared by bioconversion ginseng saponin to remove sugar )-Fine emulsified particles containing protopanaxadiol as an active ingredient through nanoemulsification technology. [Formula 1] In the above R ₁ is O-Glc ⁶ - ¹ Arap and (arabino pyrazol nosil), R ₂ is OH, R ₃ is H. The 20-O- [α-L-arabinopyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol according to claim 1, which is the active ingredient. The content of is in the range of 10 ^{-8 to} 99.99% by weight based on the total weight of the fine emulsified particles, characterized in that the fine emulsified particles. The 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol according to claim 2, which is the above-mentioned effective ingredient. The content of the fine emulsified particles is 0.001 to 30% by weight based on the total weight of the fine emulsified particles. The fine emulsified particle according to claim 1, wherein an average particle diameter of the fine emulsified particles is 30 nm to 500 nm. The fine emulsified particle according to claim 1, comprising liposomes containing lecithin, lecithin derivatives and coemulsifiers. The method of claim 1, wherein the 20-O- [α-L-arabinofyranosyl (1-> 6) -β-D-glucopyranosyl] -20 (S) -protopanaxadiol is selected from ginseng extract and A method for producing fine emulsified particles comprising ginseng purified saponin prepared by a process of acid hydrolysis, base hydrolysis or enzymatic decomposition. The method of claim 6, wherein the enzymatic digestion method is β-glucosidase, α, β-arabinosidase (α, β-arabinosidase), α, β-rhamnose which break down sugar bonds with enzymes. A method for producing microemulsified particles, characterized by using exo-sugar-binding degrading enzymes such as degrading enzymes (α, β-rhamosidase) and complex enzymes containing them. The method of claim 1, wherein the nanoemulsification technique is a high-pressure emulsification method, and the pressure condition is 500-2500 bar. The method of claim 5, wherein the content of the lecithin is 0.5 to 10% by weight based on the total weight of the finely emulsified particles, and the components of the lecithin are unsaturated choline compounds, serine compounds, and ethanolamines comprising phosphatidylcholine, lysophosphatidylcholine or phosphatidylethanolamine. Method for producing finely emulsified particles, including liposomes, characterized in that in the form of a compound or a hydrogenated system. The method of claim 5, wherein the co-emulsifier is an anionic, cationic, nonionic, or zwitterionic emulsifier, and is used at a ratio of 0.1 to 10 times the lecithin content to be used. A skin external preparation composition for preventing skin aging containing the fine emulsified particles prepared according to any one of claims 1 to 10 at a ratio of 10 ^{-8 to} 99.99% by weight. 12. The method of claim 11, wherein the skin external preparation is softening cream, astringent makeup, nourishing cosmetic, nutrition cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, body lotion, body A skin external composition comprising a cream, body oil and body essence, makeup base, foundation, hair dye, shampoo, rinse, body cleanser, toothpaste, mouthwash, ointment, patch or spray.