KR20070091443A - POLYMER-LIPOSOME COMPLEXES WITH VARIOUS HYDROPHOBIC BIOACTIVE MATERIALS SOLUBILZED BY HYDROXYPROPYL-beta;-CYCLODEXTRIN - Google Patents

Abstract

A polymer-liposome complex with various hydrophobic physiological materials solubilized by hydroxypropyl-beta-cyclodextrin is provided to secure the skin absorption promoting effect and the titer decrease preventing effect of a hydrophobic physiological active ingredient by supporting the hydrophobic physiological active ingredient in an inner aqueous phase of the complex using hydroxypropyl-beta-cyclodextrin. A polymer-liposome complex is characterized in that it comprises a liposome structure including a poly(methacrylic acid-co-n-alkyl methacrylate) random copolymer represented by the formula(1)(wherein n is from 11 to 22, and the molar ratio of x:y is 85:15-70:30) and a lipid layer, and a hydrophobic physiological active ingredient solubilized by hydroxypropyl-beta-cyclodextrin is supported at the inner phase of the liposome structure. The method comprises the steps of: (a) mixing the copolymer of the formula(1) and lipid with water and dissolving it with heating at a temperature of 50-70 deg.C; (b) mixing hydroxypropyl-beta-cyclodextrin and a hydrophobic physiological active ingredient with water and heating it at a temperature of 60-70 deg.C with solubilizing the hydrophobic physiological active ingredient using an emulsifier; (c) adjusting the pH of the solution obtained from the step(b) with an acid or a base to more than 7; (d) mixing the mixture solution obtained from the step(a) with the aqueous solution of the step(c) heated at a temperature of 50-70 deg.C and firstly dispersing it using an emulsifier; (e) nano-granulating the mixture solution obtained from the step(d) using a high pressure emulsifier; and (f) removing organic solvent used from the mixture solution obtained from the step(e) using a rotary evaporator.

Description

Polymer-Liposome Complexes with Various Hydrophobic Bioactive Materials Solubilzed by Hydroxypropyl-β-Cyclodextrin} Supporting Soluble Bioactive Components Solubilized Using Hydroxypropyl-Betacyclodextrin

1 is a schematic diagram of a polymer-liposome nanocomposite carrying a soluble bioactive component solubilized through hydroxypropyl-betacyclodextrin.

Figure 2 is a graph comparing the skin absorption of rhubarb extract through the Franz-cell of Example 2 and Comparative Example 5 of the present invention.

Figure 3 is a graph comparing the skin absorption of rhubarb extract through the Franz-cell of Example 4 and Comparative Example 8 of the present invention.

The present invention relates to a polymer-liposomal nanocomposite and a method for preparing the same, which are solubilizing useful physiologically active ingredients using hydroxypropyl-betacyclodextrin, and supporting them in an aqueous phase, specifically, methacrylic acid and n-alkyl. Polymer-liposomal nanocomposites capable of supporting a variety of useful bioactive components, including lysopropyl-betacyclodextrin solubilized using hydroxypropyl-betacyclodextrin in the water of polymer-liposome nanocomposites composed of methacrylate and lipid layers It is about.

Currently, about 40% of candidate drugs with high physiological activity that are potentially expected to be of great value cannot be entered into the development stage due to their low solubility. Solubilization is a process that enhances solubility by any manipulation of drugs that are insoluble in water or aqueous solutions. Traditional techniques for improving solubility and increasing drug absorption include the use of ethanol or surfactants as additives, the preparation of salts in the presence of ionizing groups, the method of increasing solubility by adjusting pH, etc. There is this. Recently, solubilization technology of poorly soluble drugs using liposomes, microemulsions, cyclotextrin, and the like has been advanced.

In particular, cyclodextrin has a structure in which 7 molecules of glucose are attached in a circular ring shape, and has a space of a hydrophobic environment in a molecule, and is widely used for solubilization of various types of useful drugs. In addition, the potential transdermal absorption enhancing effect of cyclodextrin has been studied. These findings have not been elucidated as to the ability of cyclodextrins as carriers through the skin, but the hypothesis is that cyclodextrins may improve the dermal absorption of drugs by increasing their thermodynamic activity.

On the other hand, one of the materials that are in the limelight as a means of gene transfer or drug delivery is liposomes. Typical lipid-cholesterol-based liposomes are safe and effective carriers composed of a thermodynamically stable lipid bilayer that surrounds the inner nucleus along with a water-soluble inner nucleus. However, lipid-cholesterol-based liposomes as drug carriers have low structural stability. In general, lipid-cholesterol-based liposomes loaded with drugs are limited in their use because of their unstable liposome structure in aqueous solutions in which various salts are used in the biological environment, or in formulations in which various surfactants are used. In addition, the use of liposomes as carriers by incorporating the soluble bioactive components into the lipid bilayer of liposomes has the disadvantage that only limited soluble bioactive components can be supported on the lipid bilayers of liposomes.

In order to overcome these drawbacks of the liposome system, there have been many attempts to overcome the above problems by utilizing the advantages of cyclodextrin and liposomes by solubilizing the useful bioactive component using cyclodextrin and then loading it in the inner water of the liposome. (B. McCormack, G. Gregoriadis, International Journal of Pharmaceutics , 162, 1998, 59-69). However, there are problems with systems using such cyclodextrins and liposomes together. In the case of general lipid-cholesterol-based liposomes, cyclodextrins used to solubilize oil-soluble bioactive components and to carry a greater amount of oil-soluble bioactive components in liposomes affect the cholesterol or lipid components that make up the liposome structure. Research has shown that it acts as an unstable element in its own structure (DG Fatouros et al., European Journal of Pharmaceutical Sciences , 13, 2001, 287-296). In addition, even though the introduction of lipid-cholesterol-based liposomes to enhance skin absorption of ketoprofen solubilized through cyclotextrin, no effect of skin absorption was observed, the carrier system using cyclodextrin and general liposomes simultaneously. Instability can be seen indirectly (F. Maestrelli et al., International Journal of Pharmaceutics , 298, 2005, 55-67).

Therefore, in order to use a liposome system carrying a useful bioactive component in various fields, a liposome without crystal precipitation of the useful bioactive component while maintaining its own structure in an aqueous solution containing cyclodextrin, various surfactants, and various salts is used. In addition to ensuring long-term stability, there is an urgent need to develop a liposome system that effectively enhances skin absorption of oil-soluble physiologically active ingredients supported at high concentrations through long-term stable liposome transporters.

Thus, the present inventors first solubilize various oil-soluble physiologically active ingredients using hydroxypropyl-betacyclodextrin in order to solve the conventional problems, and secondary for the skin absorption enhancing effect of the solubilized oil-soluble physiologically active ingredients. The research was carried out to support the polymer-liposome nanocomposite in the inner water phase. The polymer-liposomal nanocomposites used herein have the ability to carry a certain amount of water-soluble and oil-soluble physiologically active ingredients between the aqueous phase and lipid bilayers of liposomes, as well as the bioactive constituents in the last study of the present inventors. Supported polymer-liposomal nanocomposites were confirmed that the intrinsic structure of liposomes stably exist even in aqueous solutions or cosmetic formulations in which aqueous solutions or various salts exist (Patent Application No. 10-2005-0134707, patent application) 10-2005-0135409).

In order to carry a useful bioactive component that is not directly supported as a lipid bilayer of liposomes in a polymer-liposomal nanocomposite having excellent stability, a method of solubilizing a useful bioactive component using hydroxypropyl-betacyclodextrin was attempted. , The polymer-liposomal nanocomposites carrying solubilized oil-soluble bioactive components have long-term stability in aqueous solution and prevent the decrease in titer of the oil-soluble bioactive components supported. The present invention has been found and completed.

This solved the problem that various useful bioactive components in the conventional lipid-based liposome system could not be effectively loaded on liposomes, and at the same time, the safety of liposome particles without crystallization and titer reduction of useful bioactive components under any storage conditions before use. The significance of the invention is that it satisfies the need to maintain it simultaneously. In addition, the present invention was confirmed that the polymer-liposomal nanocomposite enhances the skin absorption of oil-soluble physiologically active ingredients can be said to have great utility and value.

An object of the present invention is to provide a random copolymer composed of a monomer of methacrylic acid and n-alkyl methacrylate, and a polymer-liposome nanocomposite prepared using a lipid layer as a component. The present invention provides a method of supporting various kinds of useful bioactive components solubilized with hydroxypropyl-betacyclodextrin.

Another object of the present invention has a particle size of 100 ~ 300 nm, depending on the concentration of the polymer and lipids used, and the concentration of hydroxypropyl-betacyclodextrin and oil-soluble physiologically active ingredients supported on the inner surface of the liposome, containing a polymer The polymer-liposomal nanocomposite, which maintains the same form as the lipid-based liposome, does not improve the skin absorption of the useful bioactive component.

The present invention comprises a liposome structure containing a poly (methacrylic acid-co-n-alkyl methacrylate) random copolymer of formula (1) and a lipid layer, wherein the liposomal structure has a hydroxypropyl-beta cyclodextrin on the inner It provides a polymer-liposomal nanocomposite characterized in that it carries a solubilized oil-soluble bioactive component.

In the present invention, cholesterol may be contained in the lipid layer.

In the present invention, the poly (methacrylic acid-co-n-alkyl methacrylate) random copolymer has two monomers of methacrylic acid (methacrylic acid) and stearyl methacrylate (Formula 2) It may comprise a polymer consisting of.

(In the above formula, n: m is preferably 85:15 to 70:30 relative to the molar ratio.)

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

The polymer used in the polymer-liposomal nanocomposite of the present invention is a methacrylic acid monomer introduced to impart acidity sensitivity and n-alkyl methacrylate introduced to enable association with the double lipid layer of liposomes. Acid-sensitive polymers composed of (n-alkyl methacrylate) monomers.

In general, polymers synthesized with methacrylic acid are known to cause structural changes according to the acidity change in aqueous solution. The range and degree of acidity sensitivity are determined by the number of alkyl chains and molecular weight and It is known that it depends on the polydispersity index, which is an index indicating the molecular weight distribution.

In the present invention, the n-alkyl methacrylate introduced to allow the association with the double lipid layer of liposomes has a range of alkyl chain lengths that can be associated according to the type of lipid to be associated, and generally n > 7, i.e., it is possible to associate with the double lipid layer when it has a length greater than octyl, preferably n = 11-21, and depending on the purpose, it is possible to use a separate cholesterol for the lipid bilayer or It is also possible to introduce monomers having cholesterol components.

The polymer composed of the methacrylic acid monomer and the n-alkyl methacrylate monomer is prepared by polymerization by a general free radical thermal initiation method, and may also use anionic or cationic polymerization to control molecular weight distribution. have. The mixing ratio of methacrylic acid (x in Formula 1) and n-alkyl methacrylate (y in Formula 1) used in the polymer polymerization is 90:10 to 50:50 relative to the molar ratio, preferably 85:15 to 70:30 is appropriate. The molecular weight of the polymer composed of the methacrylic acid monomer and n-alkyl methacrylate monomer affects the structure of the polymer-liposomal nanocomposite, and the molecular weight of the polymer used in the polymer-liposomal nanocomposite of the present invention is based on the number average molecular weight. 5,000 to 100,000, preferably 10,000 to 50,000.

In the present invention, in particular, a polymer of a polymer-liposome nanocomposite is composed of a methacrylic acid monomer and a stearyl methacrylate monomer introduced to enable association with a lipid bilayer of liposomes. Was used.

In the present invention, stearyl methacrylate introduced to enable the association with the lipid bilayer of liposomes can be associated with the lipid bilayer depending on the type of lipid to be associated, and depending on the purpose, a cholesterol component instead of a stearyl chain. It is also possible to introduce a monomer having

As the lipid component used to prepare the polymer-liposomal nanocomposite of the present invention, phospholipids or nitrogenous lipids having fatty acid chains having 12 to 24 carbon atoms may be used. In general, it is more preferable to use phospholipids, for example, egg yolk lecithin (phosphatidylcholine), soybean lecithin, lysolecithin, sphingomyelin, phosphatidine acid, phosphatidylserine, phosphatidylglycerol, phosphatidyl inositol Natural phospholipids selected from the group consisting of phosphatidylethanolamine, diphosphatidylglycerol, cardiolipin, and plasmalogen; Hydrogenated products of natural phospholipids; Disetylphosphate, distearoylphosphatidylcholine, dioleoylphosphatidylethanolamine, dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylethanolamine, dipalmitoylphosphatidylserine, eleostaroylphosphatidylcholine, eleostaloyl phosphatidyldilamine Synthetic lipids of astearoylphosphatidylserine; Derivatives of synthetic lipids; And fatty acid mixtures obtained by hydrolysis of synthetic lipids.

When two kinds of phospholipids are used in combination, for example, a combination of phosphatidylcholine: phosphatidylethanolamine, phosphatidylcholine: phosphatidylglycerol, phosphatidylcholine: phosphatidyl inositol, phosphatidylcholine: phosphatidine acid or phosphatidylcholine: dioleoylphosphatidylethanolamine, etc. Although the mixing ratio is different depending on the component composition to be mixed, the mixing ratio of the maximum component to the minimum component is preferably 1: 5 or less. For example, in the case of phosphatidylcholine: dioleoylphosphatidylethanolamine, in the range of molar ratios of 5: 1 to 1: 5, respectively, 1: 1, 2: 1, 3: 1, 4: 1, 5: 1, 1 : 5, 1: 4, 1: 3 or 1: 2 can be mixed in various ways. In addition, when three kinds of phospholipids are used in combination, for example, phosphatidylcholine: dioleoylphosphatidylethanolamine: phosphatidylserine in a molar ratio range of 1: 5, respectively, 1: 1: 1, 2: 1: 1, It can be mixed and used in various mixing ratios such as 3: 1: 2, 3: 2: 1, 3: 2: 2, 4: 1: 1, or 4: 2: 1.

The ratio of the polymer, cholesterol, and lipids used in the polymer-liposomal nanocomposite used in the present invention is 1 to 50%, preferably 20 to 40%, for cholesterol, and 50 to 99 for lipids, based on the weight of the composition. %, Preferably 60 to 80% is used, and in the case of polymer, 1 to 50%, preferably 5 to 30% is suitable. In addition, the content of the polymer, cholesterol, and lipids used in the polymer-liposomal nanocomposite is 0.001 to 10% by weight, and preferably 1 to 5% by weight based on the aqueous solution of the polymer-liposomal or no complex.

Hydroxypropyl-betacyclodextrin for solubilizing the useful bioactive component supported on the water phase of the polymer-liposomal nanocomposite of the present invention is 3 to 15 times the content of the useful bioactive component to be supported, preferably 5 to 5 times. Use 10 times.

In addition, the useful bioactive component solubilized by hydroxypropyl-betacyclodextrin and supported on the water phase of the polymer-liposome nanocomposite is an index component of rhubarb extract and antioxidant extract, which have antioxidant, skin stabilizing and whitening effects. Raponicin (rhaponticin), and undecylenoyl phenylalanine (undecylenoyl phenylalanine) known to have a whitening effect is contained in 0.001 to 5% by weight relative to the aqueous solution of the polymer-liposomal nanocomposite, preferably 0.1 to 2% by weight.

In general, liposomes are prepared using ultrasonic waves or using a high pressure emulsifier, and the polymer-liposomal nanocomposite according to the present invention uses a high pressure emulsifier.

a) mixing the poly (methacrylic acid-co-stearyl methacrylate) random copolymer, the following formula (1), cholesterol and lipids with an organic solvent that can be mixed with water, and dissolving the mixture at 50 to 70 ° C. to warm it;

b) mixing hydroxypropyl-betacyclodextrin with oil-soluble bioactive component in water and warming it to 60-70 ° C. to solubilize the oil-soluble bioactive component with an emulsifier;

c) adjusting the aqueous solution of b) to pH 7 or above with acid or base;

d) mixing the mixed solution of a) with an aqueous solution of c) heated to 50 to 70 ° C. and dispersing it first with an emulsifier;

e) nanoparticle-forming the mixture of d) with a high pressure emulsion; And

f) removing the used organic solvent by using the rotary evaporator of the mixed solution of e).

[Formula 1]

The organic solvent that can be mixed with water used in step a) is not limited, but is preferably selected from the group consisting of ethanol, methanol, isopropyl alcohol, acetone and tetrahydrofuran. In addition, the nano-particulation process of step e) uses a high pressure emulsifier, and can control the pressure to 100 to 1000 bar according to the desired particle size and dispersion degree, and preferably 500 to 1000 bar. In addition, the step f) may also include a step of adjusting the concentration of the polymer-liposomal nanocomposite and the useful bioactive component by removing water together.

The polymer-liposomal nanocomposite of the present invention obtained by the above production method is 100-300 nm depending on the concentration of polymer, cholesterol and lipids, and the concentration of hydroxypropyl-betacyclodextrin and oil-soluble physiologically active component supported on the liposome water phase. It has a particle size of and maintains the same particle shape as the lipid-cholesterol-based liposomes do not contain a polymer, as shown in Figure 1 is a useful bioactive component solubilized by hydroxypropyl-betacyclodextrin as the polymer-liposomal nanocomposite It will be supported on the inner water. In addition, as shown in FIG. 1, the polymer is associated with a lipid-cholesterol-based lipid bilayer to tightly bind the lipid bilayer and to protect the outer wall of the liposome, thereby stably from the elements that destabilize various liposome structures present in the water phase. It plays a role in maintaining the structure of liposomes.

The cosmetic composition containing the polymer-liposomal nanocomposite carrying the oil-soluble physiologically active component solubilized by the hydroxypropyl-betacyclodextrin of the present invention in the aqueous phase is not particularly limited in its formulation. Lotion, nourishing lotion, nourishing cream, massage cream, eye cream, eye essence, essence, cleansing cream, cleansing lotion, cleansing foam, cleansing water, pack, powder, makeup base, foundation, body lotion, body cream, body oil, It can be formulated into a body essence, body cleanser, and the like.

Hereinafter, the present invention will be described in more detail with reference to Examples and Test Examples, but the present invention is not limited only to these examples.

Examples 1 to 5 Preparation of Polymer-Liposome Nanocomposites Carrying Soluble Bioactive Components Solubilized with Hydroxypropyl-Betacyclodextrin in Water

In Examples 1 to 5, hydroxypropyl-betacyclodextrin, oil-soluble physiologically active ingredients, and acids or bases were added to the purified water at a weight composition ratio shown in Table 1 below, warmed up to 60 to 70 ° C., and then used as an homogenizer. The mixture is stirred for 10 minutes at a speed of 7000 rpm to solubilize the useful bioactive component. At the same time poly (methacrylic acid-co-stearyl methacrylate) copolymer (mol composition 70:30, number average molecular weight 26,000), cholesterol and 100% hydrogenated oleorel-palmitoyl / oleoreyl-stearyl phosphatidyl The choline mixture (lipoid S100-3) was warmed and dissolved in an organic solvent at 60 to 70 ° C. at a weight composition ratio shown in Table 1 to prepare a mixed solution. The above-mentioned organic solvent mixture was added to an aqueous solution in which the oil-soluble physiologically active component was solubilized, and heated to 60-70 ° C., followed by mixing and stirring for 5 minutes at a speed of 5000 rpm with an emulsifier, solubilizing the soluble physiologically active component with hydroxypropyl-betacyclodextrin. After preparing a polymer-liposomal complex supported on the aqueous phase was nanoparticles using a high pressure homogenizer (1000bar, 3cycle). Residual organic solvent was removed using a rotary evaporator, and in some cases, a certain amount of water was also distilled out to adjust the concentrations of the useful bioactive component and the polymer-liposomal nanocomposite.

[Comparative Examples 1 to 3] Preparation of Polymer-Liposome Nanocomposites Directly Supporting Soluble Bioactive Components Without Using Hydroxypropyl-Betacyclodextrin

For Comparative Examples 2 to 3 poly (methacrylic acid-co-stearyl methacrylate) copolymer (mol composition 70: 30, number average molecular weight 26,000), cholesterol, 100% hydrogenated oleyl-palmitoyl / ol Leoyl-stearyl phosphatidyl choline mixture (lipoid S100-3) and oil-soluble physiologically active ingredients are dissolved in an organic solvent of 60 ~ 70 ℃ at the weight composition ratio shown in Table 2 to prepare a mixed solution. The organic-solvent mixture is mixed with water containing an acid or a base at 60-70 ° C., and mixed by stirring for 5 minutes at a speed of 5000 rpm with an homogenizer, thereby polymer-liposomes containing the first dispersed useful bioactive component. After preparing the composite was nanoparticles using a high pressure homogenizer (1000bar, 3cycle). Residual organic solvent was removed using a rotary evaporator, and in some cases, a certain amount of water was also distilled out to adjust the concentrations of the useful bioactive component and the polymer-liposomal nanocomposite.

In Comparative Example 1, except for using the poly (methacrylic acid-co-stearyl methacrylate) copolymer (molecular composition 85:15, number average molecular weight 20,000) in the same manner as in the above Comparative Examples 2 to 3 An acidity sensitive polymer-liposomal nanocomposite containing the active ingredient was prepared.

[Comparative Examples 4 to 7] Preparation of an aqueous solution in which an oil-soluble physiologically active component was solubilized using hydroxypropyl betacyclodextrin

In Comparative Examples 4 to 7, hydroxypropyl-betacyclodextrin, oil-soluble physiologically active ingredient, and acid or base were added to the purified water at the weight composition ratio shown in Table 3 below, and then heated to 60-70 ° C., followed by an emulsifier. The mixture was stirred for 10 minutes at a speed of 7000 rpm to prepare an aqueous solution in which the oil-soluble bioactive component was solubilized.

Constituents and Compositions of Soluble Bioactive Components Solubilized with Hydroxypropyl-Betacyclodextrin Active ingredient Composition (g) Active ingredient Hydroxypropyl-Betacyclodextrin Acid or base water Comparative Example 4 Lafonticin One 5 0 94 Comparative Example 5 Lafonticin 2 10 0 88 Comparative Example 6 Rhubarb extract One 5 Sodium Phosphate (Dibasic) 0.032 93.97 Comparative Example 7 Undecylenoyl Phenylalanine One 5 0 94

Comparative Example 8 Preparation of Nanoemulsion Supporting 1% by Weight of Rhubarb Extract

In order to observe the decrease in titer and skin absorption comparison of the polymer-liposomal nanocomposite and rhubarb extract carrying 1 wt% rhubarb extract prepared in Example 4, rhubarb in the weight composition ratio shown in Table 4 below A nanoemulsion carrying a weight% was prepared.

Rhubarb Extract-supported Nanoemulsion Components and Contents Raw material name Content (% by weight) Rhubarb extract One Polyglycol E400 NF 10 Hydrolite-5 10 Ethanol 95% 10 General R E5 2 Lipoid S 75-3 4 Purified water 62.9 E.D.T.A.-2NA 0.05 Aminofect 0.05

Experimental Example 1 Observation of Particle Size of Polymer-Liposome Nanocomposites Supported by Soluble Bioactive Components

Without using the polymer-liposomal nanocomposites in which the oil-soluble physiologically active component solubilized with the hydroxypropyl-betacyclodextrins of Examples 1 to 5 was supported on the water and the hydroxypropyl-betacyclodextrins of Comparative Examples 1 to 3, The particle size of the polymer-liposomal nanocomposites directly loaded with oil-soluble bioactive components was measured using a dynamic light scattering apparatus (Zetasizer 3000HSa Malvern Instruments).

Result of particle size measurement of polymer-liposomal nanocomposites dispersed in aqueous solution. Particle Size (nm) Particle Size (nm) Example 1 132 Comparative Example 1 Sedimentation and Float Generation Example 2 115 Comparative Example 2 Sedimentation and Float Generation Example 3 195 Comparative Example 3 Sedimentation and Float Generation Example 4 169 Comparative Example 4 - Example 5 147 Comparative Example 5 - - - Comparative Example 6 -

As shown in Table 5, the polymer-liposomal nanocomposites of Examples 1 to 6 were able to obtain a stable liposome aqueous solution having a specific particle size of 115 to 219 nm. On the other hand, in the case of the polymer-liposomal nanocomposites of Comparative Examples 1 to 3 shown in Table 5, an aqueous solution of the polymer-liposomal nanocomposite having a specific particle size was not obtained, and the polymer-liposomal nanocomposite together with the crystallization of the useful bioactive component. Was found not to be formed properly. In addition, it can be seen that the particle size of the polymer-liposomal nanocomposite is different depending on the type of the soluble bioactive component solubilized by hydroxypropyl-betacyclodextrin and the concentration of the component of the polymer-liposomal nanocomposite. . Polymer-liposome nanocomposite components in which the organic solvent mixture in which the components of the polymer-liposome nanocomposites are dissolved is added to the aqueous solution in which the soluble physiologically active component solubilized with hydroxypropyl-betacyclodextrin is dispersed and mixed and stirred. The concentration of is thought to have the greatest effect on particle size.

On the other hand, in the case of the useful bioactive component solubilized with hydroxypropyl-betacyclodextrin in the aqueous solution of Comparative Examples 4 to 6 shown in Table 5, no crystallization or suspended solids were observed, and a clear and clear aqueous solution having a specific color was used. Could get This means that the oil-soluble physiologically active component is dispersed in an aqueous solution at the molecular level by hydroxypropyl-betacyclodextrin.

Test Example 2 Evaluation of Particle Size Long-term Stability of Polymer-Liposome Nanocomposites Carrying Soluble Bioactive Components Soluble with Hydroxypropyl-Betacyclodextrin

A polymer-liposomal nanocomposite aqueous solution prepared in order to observe the particle size long-term stability of the polymer-liposomal nanocomposite in which the oil-soluble physiologically active component solubilized with the hydroxypropyl-betacyclodextrin of Examples 1 to 4 was carried out in the water (2 ° C), room temperature (25 ° C) and high temperature (40 ° C), respectively, to store the change in particle size with time using a dynamic light scattering device (Dynamic light scattering, Zetasizer 3000HSa Malvern Instruments) 6 is shown.

Long-term stability measurement of particle size of polymer-liposomal nanocomposites dispersed in aqueous solution Particle size (nm) At the time of manufacture 1 week 2 weeks 4 weeks 8 Weeks 12 Weeks 16 Weeks Example 1 2 ℃ 132 133 135 137 137 140 139 Room temperature 134 139 140 145 144 143 40 ℃ 134 142 146 153 148 154 Example 2 2 ℃ 115 111 117 112 114 120 114 Room temperature 120 119 126 120 122 122 40 ℃ 119 126 124 126 131 134 Example 3 2 ℃ 195 197 197 201 203 201 204 Room temperature 198 201 208 206 204 206 40 ℃ 197 206 218 212 217 220 Example 4 2 ℃ 169 172 171 170 165 171 173 Room temperature 170 174 172 172 175 177 40 ℃ 172 176 175 182 189 187

As shown in Table 6, when the polymer-liposomal nanocomposites of Examples 1 to 4 were stored at 2 ° C. and 25 ° C., no significant change in particle size with time was observed.

On the other hand, it was observed that the particle size of the polymer-liposomal nanocomposites of Examples 1 to 4 stored at 40 ° C. was relatively higher than that at other temperatures compared to the particle size at the time of manufacture, which was at a high temperature of 40 ° C. The self-association of the lipid components constituting the lipid bilayer of the polymer-liposomal nanocomposite is caused by the expansion of the polymer-liposomal nanocomposite that occurs while being relatively loose compared to the low temperature and room temperature of 2 ° C and 25 ° C.

In addition, the particle size of the polymer-liposomal nanocomposite present in the aqueous solution together with the hydroxypropyl-betacyclodextrins of Examples 1 to 4 was maintained for a long time without large change to the cyclodextrin of the polymer-liposome nanocomposite. The structural stability was confirmed. In the polymer-liposomal nanocomposite, the polymer acts as a supporter that tightly binds the liposome's lipid bilayer, so that even in the presence of cyclodextrin, it is possible to maintain a stable liposome intrinsic structure compared to existing lipid-cholesterol-based liposomes.

Experimental Example 3 Observation of rhubarb extract titer in a polymer-liposome nanocomposite in which the rhubarb extract solubilized by hydroxypropyl-betacyclodextrin was supported in the aqueous phase and a nanoemulsion in which the rhubarb extract was supported

The potency of the rhubarb extract of the rhubarb extract in the nanoemulsion in which the rhubarb extract solubilized with the hydroxypropyl-betacyclodextrin of Example 4 supported on the aqueous phase and the rhubarb extract of Comparative Example 8 was observed. The samples of Example 4 and Comparative Example 8 were stored at room temperature and 40 ° C., respectively, and the results of measuring the concentration of raponticin, which is an index component of rhubarb extract, over time using a high performance liquid chromatography, It is shown in Table 7 below.

Initial potency 1 week 8 Weeks 16 Weeks Example 4 Room temperature 100% 100% 100% 100% 40 ℃ 100% 100% 100% 100% Comparative Example 8 Room temperature 100% 100% 100% 98.4% 40 ℃ 100% 97.7% 97.3% 87.7%

As shown in Table 7, no decrease with time was observed at both the room temperature and 40 ° C. at the concentration of lafonticin of Example 4. On the other hand, the concentration of the raponticin of Comparative Example 8 was found to show more decrease in titer at 40 ° C. than at room temperature over time. Raponicin, an index component of rhubarb extract, is a useful bioactive component that tends to decrease its titer by light or temperature. According to the results of Table 7, the titer stability of the rhubarb extract supported on the inner water of the polymer-liposomal nanocomposite after collecting and solubilizing by hydroxypropyl-betacyclodextrin was higher than that of the rhubarb extract supported on the nanoemulsion. It can be assumed that the rhubarb extract is trapped in the hydroxypropyl-betacyclodextrin used for solubilizing the rhubarb extract, rather than stabilization by the polymer-liposomal nanocomposite, thereby preventing the decrease in the titer of rapontisine.

[Test Example 4] Comparative experiment of skin absorption of rhubarb extract through Franz-cell

A polymer-liposome nanocomposite that supports solubility in hydroxypropyl-betacyclodextrin solubilized with hydroxypropyl-betacyclodextrin prepared in Examples 2 and lafontisine solubilized in hydroxypropyl-betacyclodextrin, respectively. The skin absorption of raponticin in aqueous solution was compared. In addition, the skin of the rapontisin of the nanoemulsion which is carrying the polymer-liposome nanocomposite carrying the rhubarb extract solubilized with hydroxypropyl-betacyclodextrin prepared in Examples 4 and 8 and the rhubarb extract respectively. The absorption was compared. For skin absorption experiment, skin absorption experiment was performed for 18 hours using Franz-cell device using the skin extracted from albino guinea pig, and raponti absorbed by skin to compare the degree of skin absorption of rapontisin. The concentration of the scene was measured using a high performance liquid chromatography (high performance liquid chromatography) is shown in Figures 2 and 3.

FIG. 2 shows a polymer-liposome nanocomposite in which rapponicine solubilized by the hydroxypropyl-betacyclodextrin of Example 2 and rapponticin solubilized by the hydroxypropyl-betacyclodextrin of Comparative Example 5 This is a result of comparing the skin absorption of rapontisin in the dispersed aqueous solution. Rapontisin solubilized by hydroxypropyl-betacyclodextrin also showed a significant amount of skin absorption. In addition, it was confirmed that 162% of Rapontisin skin absorption was enhanced by the polymer-liposomal nanocomposite. The mechanism of enhancing skin absorption by cyclodextrin or liposome system has not been elucidated, but many experiments have already shown the effect. However, since the structural instability of liposomes occurs in the existing lipid-cholesterol-based liposomes due to the presence of cyclodextrins, the existing studies are expected to enhance skin absorption by both cyclodextrin and liposome systems as shown in FIG. 2. It was difficult. In the polymer-liposomal nanocomposite, the polymer acts as a supporter that tightly binds the lipid bilayer of liposomes, thereby maintaining the intrinsic structure of the liposome even in the presence of cyclodextrin, thereby promoting skin absorption.

3 is an indicator component of rhubarb extract in a nanoemulsion in which the rhubarb extract solubilized by the hydroxypropyl-betacyclodextrin of Example 4 was loaded on the water and the rhubarb extract of Comparative Example 8; This is a result of comparing skin absorption of inrapponticin. As can be seen from FIG. 3, it was found that 842% of the increase in skin absorption of rapontisin in the rhubarb extract of the polymer-liposomal nanocomposite of Example 4 compared to the rhubarb extract supported on the nanoemulsion. Since it shows much higher skin absorption effect of rapontisin than the conventional formulation, it is very helpful in formulating various formulations of useful bioactive components through hydroxypropyl-betacyclodextrin and high stability polymer-liposomal nanocomposite. Could be.

As described above, the polymer-liposomal nanocomposites carrying the soluble physiologically active component solubilized with hydroxypropyl-betacyclodextrin according to the present invention in the aqueous phase are incorporated into the lipid bilayer of conventional lipid-cholesterol-based liposomes. Various useful physiologically active ingredients, which were impossible, can be stably supported on the inner water of the polymer-liposomal nanocomposite having excellent stability, and prevent the decrease in titer of the soluble physiologically active ingredients supported thereon as well as enhance skin absorption.

In addition, it was confirmed that the polymer-liposomal nanocomposite according to the present invention was relatively stable with respect to cyclodextrins compared to conventional lipid-cholesterol-based liposomes, and through this, polymer-liposomal nanocomposites were used together with cyclodextrins. It has much improved stability against various salts and cosmetic formulations than the well-known lipid-cholesterol-based liposomes, and it is also very useful because it has excellent skin absorption enhancement effect.

Claims (11)

A poly (methacrylic acid-co-n-alkyl methacrylate) random copolymer of formula (I) comprising a liposome structure containing a lipid layer and a solubilized solubility with hydroxypropyl-beta cyclodextrin on the inside of the liposome structure A polymer-liposomal nanocomposite comprising a bioactive component. [Formula 1]

The polymer-liposomal nanocomposite of claim 1, wherein cholesterol is incorporated into the lipid layer. 3. The poly (methacrylic acid-co-n-alkyl methacrylate based on the total weight of the poly (methacrylic acid-co-n-alkyl methacrylate) random copolymer, cholesterol and lipids according to claim 2 A polymer-liposomal nanocomposite comprising 1 to 50% by weight of random copolymer, 50 to 99% by weight of lipid and 1 to 50% by weight of cholesterol. The polymer-liposome nanocomposite of claim 1, wherein the poly (methacrylic acid-co-n-alkyl methacrylate) random copolymer has a number average molecular weight of 10,000 to 50,000. According to claim 1, wherein the lipid is a yolk lecithin (phosphatidylcholine), soybean lecithin, lysolecithin, sphingomyelin, phosphatidyl acid, phosphatidylserine, phosphatidyl, having a fatty acid chain of 12 to 24 carbon atoms Natural phospholipids selected from the group consisting of glycerol, phosphatidyl inositol, phosphatidylethanolamine, diphosphatidylglycerol, cardiolipin and plasmalogen; Hydrogenated products obtainable from natural phospholipids; Dicetylophosphate, distearoylphosphatidylcholine, dioleoyl phosphatidylethanolamine, dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylethanolamine, dipalmitoylphosphatidylserine, eleostaroyl phosphatidylcholine, eleostaroylphosphamine and eldiethanol Synthetic lipids selected from the group consisting of astearoyl phosphatidylserine; Derivatives of synthetic lipids; And a fatty acid mixture obtainable by hydrolysis of synthetic lipids. The polymer-liposomal nanocomposite according to claim 1 or a mixture of two or more selected from the group consisting of. The polymer-liposomal nanocomposite of claim 1, wherein the oil-soluble bioactive component is contained in an amount of 0.001 to 5% by weight based on the entire polymer-liposomal nanocomposite. The polymer-liposome nanocomposite of claim 1, wherein the oil-soluble bioactive component is selected from the group consisting of raponticin, rhubarb extract, and undecylenoyl phenylalanine. The polymer-liposome according to claim 1, wherein the hydroxypropyl betacyclodextrin used to solubilize the oil-soluble physiologically active component in an aqueous solution is contained at 3 to 15 times the content of the oil-soluble physiologically active component. Nanocomposites. The polymer-liposomal nanocomposite of claim 1, wherein the polymer-liposomal nanocomposite has a particle size of 100 to 300 nm. a) mixing the poly (methacrylic acid-co-stearyl methacrylate) random copolymer and the lipid of Formula 1 with an organic solvent that can be mixed with water to dissolve it at 50 to 70 ° C., b) mixing hydroxypropyl-betacyclodextrin with oil-soluble bioactive component in water and warming it to 60-70 ° C. to solubilize the oil-soluble bioactive component with an emulsifier; c) adjusting the solution of b) to pH 7 or above with acid or base; d) mixing the mixed solution of a) with an aqueous solution of c) heated to 50 to 70 ° C. and dispersing it first with an emulsifier; e) nanoparticle-forming the mixture of d) with a high pressure emulsion; And f) removing the organic solvent used by using the rotary evaporator in the mixed solution of e); and a polymer supporting an aqueous soluble bioactive component solubilized with hydroxypropyl-betacyclodextrin in an aqueous phase. -Preparation method of liposome nanocomposite. [Formula 1]

The method of claim 10, wherein the organic solvent that can be mixed with water in step a) is selected from the group consisting of ethanol, methanol, isopropyl alcohol, acetone, and tetrahydrofuran.

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