KR100465976B1 - Nanoemulsion having Ginsenoside F1 by nano-emulsion technology, and skin care and pharmaceutical compositions for external applications utilizing thereof - Google Patents

Abstract

The present invention is a ginsenoside F1 (20-O-β-D-glucopyranosyl-20 (S) -protopanaxatriol, which is a major metabolite of ginseng prepared by the bioconversion method, The present invention relates to a skin external preparation for maximizing skin absorption by containing the ginsenoside F1 in fine emulsified particles and liposomes, using 'ginsenoside F1' as a main active ingredient, and using nanoemulsification technology. More specifically, by using nanoemulsification technology such as high pressure emulsification technique and solvent extraction method, emulsifiers such as lecithin and emulsifiers having excellent skin-friendly emulsifiers and the method of containing the ginsenoside F1 in fine emulsified particles and liposomes The present invention relates to a skin anti-aging cosmetic and pharmaceutical composition having a greatly improved absorption rate and having excellent skin cell proliferation and collagen biosynthesis promoting effects.

Description

Nanoemulsion having Ginsenoside F1 by nano-emulsion technology, and skin care and pharmaceutical compositions for external applications utilizing

The present invention is a ginsenoside F1 (20-O-β-D- glucopyranosyl-20 (S) -protopanaxatriol, which is a major metabolite of ginseng prepared by the bioconversion method; Hereinafter referred to as ginsenoside F1) using the nanoemulsification technology in the fine emulsified particles and liposomes to the external composition for skin maximized skin absorption, more specifically nanoemulsification such as high pressure emulsion and solvent extraction method Method to contain ginsenoside F1 in fine emulsified particles and liposomes together with emulsifiers such as lecithin and skin-friendly emulsifiers, and by formulating them, greatly improves transdermal absorption and promotes excellent skin cell proliferation and collagen biosynthesis. It relates to a skin anti-aging cosmetics and pharmaceutical compositions. In addition, in formulating ginsenoside F1, in addition to the fine emulsified particles and liposomes, they 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 they are, or encapsulated. Means all methods for applying the efficacy of ginsenoside F1 as a cosmetic and a medicine by applying a microemulsion technique, including the general method of the formulation technology.

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 are being 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 It was developed in the direction of preparing and separating and purifying ginsenoside F1.

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 it was an oleanine-based saponin in order of the 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 the ginseng saponin as a raw material of the anti-aging product, it was confirmed that the efficacy of the ginsenoside F1 of the bioconversion ginsenoside F1 that maintains the efficacy of the ginseng saponin, and especially increased skin permeability.

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 sugars are linked by ether bonds to alcoholic OH groups at the R1, R2 and R3 positions in the dammaran type, and thus have high hydrophilicity, 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 and compound K, which are composed of a sugar (glucose) attached to aglycone, are known to have pharmacological effects such as inhibiting cancer cell proliferation, inhibiting tumor growth, and increasing anticancer activity of anticancer drugs. ought.

Although research on ginsenoside F1 obtained by removing a part of sugar from ginseng saponin has been actively conducted, there has been no research on the technique of introducing it into the skin and formulation technology. In addition, there is no limitation on the formulation technology used to apply the physiological activity effect of ginsenoside F1, but general emulsified particles of 500nm to 1000um or less using general emulsification technology and microemulsion particles of 500nm or less to 30nm or more using microemulsification technology, It is expected that ginsenoside F1 may be captured and encapsulated.

Therefore, in the present invention, ginsenoside F1 separated and purified from ginseng saponin by the bioconversion method as a main active ingredient and excellent skin cell proliferation by containing fine emulsified particles and liposomes inside using skin-friendly emulsifier and nanoemulsification technology. And to provide a skin anti-aging cosmetics and pharmaceutical composition having a collagen biosynthesis promoting effect.

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 relates to a composition for external application for skin that maximizes skin absorption by containing ginsenoside F1, a major metabolite of ginseng manufactured by bioconversion, in fine emulsified particles and liposomes using nanoemulsification technology. More specifically, by using nanoemulsification techniques such as high pressure emulsification and solvent extraction, a method of containing ginsenoside F1 in fine emulsified particles together with emulsifiers having excellent skin affinity such as lecithin, and formulating the same, greatly increases transdermal absorption. It relates to a skin anti-aging cosmetics and pharmaceutical compositions having improved skin cell proliferation and collagen biosynthesis promoting effect.

The ginsenoside F1 is represented by the following formula (1).

When ginsenoside F1 of Formula 1 is expressed as a general formula, it is as follows. Wherein R ₁ is O-Glc, R ₂ is OH, and R ₃ is OH.

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 above, ginsenoside F1 used in the present invention is characterized by reducing the molecular weight by removing a portion of the sugar from the common ginseng saponin, and is hydrophobic.

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

Ginsenoside F1 used in the present invention is prepared by hydrolyzing ginseng purified saponin with acid, alkali or enzyme, and removing the sugar from ginseng saponin 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, ginsenoside F1 in the preparation of fine emulsified particles and liposomes 10 relative to the total weight of the emulsified particles and liposomes ^It is contained in an amount of ^{-10 to} 70% by weight, preferably 0.001 to 30% by weight. In addition, the fine emulsified particles containing ginsenoside F1 is contained in an amount of 10 ^{-10 to} 99.99% by weight based on the total weight of the cosmetic composition, although there is a difference depending on the manufacturing method. In addition to applying the method of collecting the inside of the fine emulsified particles in the emulsified particles and capsules by a general emulsification method having a particle size of 500um to 1000um, or to dissolve in an oil capable of dissolving ginsenoside F1 It may be.

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. It increases the percutaneous absorption rate of ginsenoside F1.

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, fine emulsified particles containing ginsenoside F1 are prepared using a high pressure emulsification method (pressure 500-2,500 bar) and a solvent extraction method as nanoemulsification technology.

Skin anti-aging cosmetics and pharmaceutical compositions containing ginsenoside F1 according to the present invention is not particularly limited in the 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 l of ethanol containing water was added to 2 kg of red ginseng, and refluxed and extracted three times at 77 ° C., followed by immersion 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 Preparation of Ginsenoside F1 by Acid Hydrolysis Method

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 = 0.65) was used to isolate and recover the fraction of ginsenoside F1 to finally give 2400 mg of ginsenoside F1 (yield 2.4%).

Reference Example 3 Preparation of Ginsenoside F1 by Base Hydrolysis Method

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 = 0.65) was used to isolate and recover the fraction of ginsenoside F1 to finally give 2700 mg (g. 2.7%) of ginsenoside F1.

[Reference Example 4-1] Preparation of ginsenoside F1 through enzymatic digestion

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 = 0.65) was used to isolate and recover the fraction of ginsenoside F1 to finally give 7600 mg (ginseng 7.6%) of ginsenoside F1.

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 = 0.65) was used to isolate and recover the fraction of ginsenoside F1 to finally obtain 2600 mg of ginsenoside F1 (yield 2.6%).

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 = 0.65) was used to isolate and recover the fraction of ginsenoside F1 to finally give 1800 mg (yield 1.8%) of ginsenoside F1.

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 = 0.65) was used to separate and recover the fraction of ginsenoside F1 to finally give 2800 mg (yield 2.8%) of ginsenoside F1.

Nano-emulsified particles containing ginsenoside F1 obtained from the above reference example were prepared as in Examples 1 to 3.

Example 1

Ginsenoside F1 dissolved in lecithin, hydrogenated lecithin, cholesterol, soybean oil and propylene glycol was dissolved completely by heating to 70 ~ 75 ℃, then mixed with pre-heated water part (distilled water, EDTA) and pre-emulsified by general homomixer. Pre-emulsion was carried out (3 min, 3,000 ~ 6,000rpm) and 1,000Bar / 3cycles were carried out using a microfluidizer. In the above, hydrogenated lecithin is excellent in emulsion stability, but skin affinity in comparison with unsaturated lecithin in terms of skin absorption, was used by mixing two lecithins.

Example 2

Ginsenoside F1 dissolved in lecithin, PEG-5 rapeseed sterols, capric / caprylic triglycerides, BHT, a-tocopherol and pentylene glycol and ethanol was heated to 70-75 ° C. to complete dissolution. Pre-emulsion was mixed with a pre-heated water part (distilled water, EDTA) using a general homomixer (3,000 ~ 6,000rpm for 3 minutes) and 1,000bar / 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 ginsenoside F1 dissolved in propylene glycol and ethanol were dissolved completely by heating to 70-75 ° C, and then mixed with pre-heated water-parts (distilled water, EDTA, glycerin, betaine) The mixture was emulsified with a general homomixer (3,000 to 6,000 rpm for 3 minutes) and 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.

Comparative Example 1 to prepare the fine emulsified particles and liposomes containing the ginsenoside F1 and the general emulsion prepared in Examples 1 to 3, and to compare the percutaneous absorption difference of each ginseng extract saponin extract To Comparative Example 3 was prepared, the contents of which are summarized in Table 1 below. 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 Ginsenoside F1 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, the external composition for skin containing the fine emulsified particles and liposomes or the general emulsified particles containing ginsenoside F1 will be described.

[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 collected at regular time intervals, and then the amount of ginsenoside F1 absorbed into the skin was measured using HPLC, which is expressed as the skin absorption amount (µg / cm 2 / wt%) per application concentration. The results are shown in Table 2 below. For ginseng purified saponin, the content of transdermal absorbed saponin was quantified, and for ginsenoside F1, the total transdermal absorption was calculated by quantitative analysis of the content of transdermal absorbed ginsenoside F1.

<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 15.45 39.74 58.43 One 0 3.52 8.11 14.41 2 0 13.02 38.64 55.27 2 0 3.66 8.35 14.06 3 0 12.59 33.55 45.32 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 34.00 54.21 One 0 2.51 3.98 4.68 2 0 11.21 21.63 51.64 2 0 1.15 2.62 3.86 3 0 4.21 17.76 22.44 3 0 0.48 1.01 2.03 4 0 15.98 37.86 57.93 4 0 2.62 3.21 5.93 5 0 13.05 31.31 52.53 5 0 2.24 3.53 5.92 6 0 2.24 13.75 25.14 6 0 0.45 1.97 1.97 7 0 14.30 38.59 59.97 7 0 3.23 4.84 6.83 8 0 10.15 31.65 52.35 8 0 1.56 3.47 5.31 9 0 2.23 13.65 25.04 9 0 1.50 1.03 2.11 10 0 14.21 32.25 53.23 10 0 2.33 5.13 7.01 11 0 10.22 33.85 52.52 11 0 1.56 2.73 4.32 12 0 2.12 12.36 25.37 12 0 0.49 1.11 2.23 13 0 12.13 35.99 57.83 13 0 2.45 4.10 6.02 14 0 9.23 33.87 55.16 14 0 1.12 3.54 6.63 15 0 2.23 13.45 25.67 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 ginsenoside F1 is applied to the nanoemulsification technology (Examples 1 to 2) it was confirmed that there is a dramatic increase in transdermal absorption.

In addition, through the corresponding examples and comparative examples, it can be seen that ginsenoside F1 has a higher transdermal absorption rate than other glucosyl ginsenosides of ginseng saponin, which is a chemical of ginsenoside F1. It can be seen that it originates from the characteristic of the phosphorus structure.

In other words, ginsenoside F1 shows a much higher transdermal absorption rate than ginseng purified saponin, and in particular, by applying the lecithin and nanoemulsification technology excellent in skin affinity, it can be seen that the 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 dermal transdermal ginsenoside F1 due to the use of lecithin having excellent nanoemulsification technology and skin affinity. It was confirmed that the water absorption was reproduced and applied as it is in the formulation.

Considering that the makeup duration is generally 4 to 8 hours, it can be seen that the percutaneous absorption in the Examples and Formulations containing ginsenoside F1 is superior to the Examples and Formulations containing ginseng purified saponin. .

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 bovine serum were dispensed to 96 cells / well in a 96-well microtiter plate, Example 1 as a sample. Ginsenoside F1 of, the ginseng purified saponin of Comparative Example 1 was used in an amount of 1% each, and Formulation Example 1 and Comparative Formulation Example 1 were each used in an amount of 10% dilution sequentially 1/10 with a culture medium After the addition, the cells were incubated at 37 ° C for 4 days. After incubation, 50 μl of 0.2% MTT (3- [4,5-dimethy-lthiazol-2-yl] -2,5-diphenyltetrazolium bromide) solution was added to each well, followed by incubation at 37 ° C. for 4 hours. The resulting 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. After comparing the absorbance of the test group containing ginseng purified saponin and ginsenoside F1 and the control group not containing them, 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 10 5 8 5 1 × 10 ^-6 25 8 22 7 1 × 10 ^-5 43 12 39 11 1 × 10 ^-4 75 15 65 15 1 × 10 ^-3 95 22 85 22 1 × 10 ^-2 121 33 102 31 1 × 10 ^-1 153 41 123 39

From Table 3, it was found that the fibroblasts treated with ginsenoside F1 had a much higher proliferative effect than the fibroblasts treated with ginseng purified saponin. 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 keratinocytes in Example 2, Comparative Example 2, 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 (%) Proliferative capacity of keratinocytes (%) Example 2 Comparative Example 2 Formulation Example 2 Comparative Formulation Example 2 1 × 10 ^-8 5 4 5 3 1 × 10 ^-7 19 6 14 5 1 × 10 ^-6 38 7 24 7 1 × 10 ^-5 51 11 38 11 1 × 10 ^-4 69 14 46 14 1 × 10 ^-3 81 19 58 18 1 × 10 ^-2 101 23 72 21 1 × 10 ^-1 125 28 89 25

As can be seen in Table 4, compared to the keratinocytes treated with ginseng purified saponin, keratinocytes treated with ginsenoside F1 showed about two 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 day 3 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. Ci was added. After 24 hours, the medium and cells in each well were scraped, washed in 5% Trichloroacetic acid (TCA) solution, and dispensed into two test tubes. 1 unit / μl of type I collagenase was added thereto 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 6 2 5 2 1 × 10 ^-7 16 2 11 2 1 × 10 ^-6 21 4 18 4 1 × 10 ^-5 35 6 29 6 1 × 10 ^-4 43 10 38 9 1 × 10 ^-3 59 12 51 11 1 × 10 ^-2 71 16 66 15 1 × 10 ^-1 85 20 77 19

From the results of Table 5, compared to fibroblasts treated with ginseng purified saponin, fibroblasts treated with ginsenoside F1 showed about 3 times improved collagen biosynthesis promoting effect, and these results showed nanoemulsification technology. Even when not used, the same trends were observed in the examples prepared using ginsenoside F1 and skin-friendly lecithin and formulations containing the same.

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

Hairless mice (42 weeks, female) were coated with ginsenoside F1 (excipient; EtOH: PG = 7: 3), patched for 3 days, allowed to rest 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, it can be seen that can effectively deliver the ginsenoside F1 into the skin using skin-friendly lecithin and nanotechnology. have.

As a result, ginseng purified 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, by using skin-friendly lecithin and nanoemulsification technology in the transdermal absorption of ginsenoside F1 having the structure of easy transdermal absorption, maximized transdermal absorption is possible, and as a result, effective collagen biosynthesis It could be confirmed that this can be done.

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 72 ± 15% Comparative Formulation Example 1 23 ± 10%

As can be seen from the above results, in the present invention, in order to overcome the disadvantages of low transdermal absorption rate due to the structural characteristics of ginseng saponin, fine emulsified particles using a skin-friendly emulsifier and a nanoemulsification technique of ginsenoside F1 from which sugar was removed It was found to maximize the percutaneous absorption rate by containing it inside, and it was found that the skin aging effect such as wrinkle reduction is excellent.

Claims (12)

Ginsenoside F1 represented by the following formula (2) prepared by bioconversion ginseng saponin to remove the sugar as an active ingredient, fine emulsified particles having a particle size of 30 ~ 500nm: [Formula 2] Wherein R ₁ is O-Glc, R ₂ is OH, and R ₃ is OH. The fine emulsified particle according to claim 1, wherein the content of ginsenoside F1 as the active ingredient is 10 ^{-10 to} 70 wt% based on the total weight of the fine emulsified particles. The fine emulsified particle according to claim 2, wherein the content of ginsenoside F1 as the active ingredient is 0.001 to 30% by weight based on the total weight of the fine emulsified particles. delete The fine emulsified particle according to claim 1, wherein the fine emulsified particles are in the form of liposomes. Preparing ginsenoside F1 represented by the following Chemical Formula 2 through the process of acid hydrolysis, base hydrolysis or enzymatic digestion of ginseng extract and ginseng purified saponin: [Formula 2] Wherein R ₁ is O-Glc, R ₂ is OH, and R ₃ is OH; And Preparing nanoparticles in units of 30 to 500 nm by emulsifying the ginsenoside F1 at a pressure of 500 to 2,500 bar using lecithin or hydrogenated lecithin as an emulsifier; Method of producing fine emulsified particles comprising a. According to claim 6, β-glucosidase, α, β-arabinosidase (α, β-arabinosidase), α, β-rhamnose to decompose sugar bonds as enzymes in the enzymatic digestion method A method for producing microemulsified particles, which is prepared using an exo sugar-binding degrading enzyme such as α, β-rhamosidase or a mixed enzyme thereof. delete 7. The unsaturated choline compound, serine compound and ethanolamine according to claim 6, wherein the content of the lecithin is 0.5 to 5% by weight based on the total weight of the fine emulsified particles, and the components of the lecithin are composed of 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. 7. The method of claim 6, wherein an anionic, cationic, nonionic or zwitterionic emulsifier is used in an amount of 0.5 to 5 times the lecithin content in addition to the emulsifier. A skin external preparation composition for preventing skin aging containing the fine emulsified particles according to any one of claims 1 to 5 at a ratio of 10 ^{-10 to} 99.99% by weight. The method of claim 11, wherein the skin external preparation is a softening cream, astringent makeup, nourishing cream, nutrition cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, makeup base, A composition for external application for skin, characterized in that it is formulated as a foundation, hair dye, shampoo, rinse, body cleanser, toothpaste, mouthwash, ointment, patch or spray.