KR100654102B1 - Preparation of ph-responsive polymer-liposome complexes with various hydrophobic bioactive materials and the composition of skin external application containing the same - Google Patents

Abstract

Provided are a polymer-liposome nanocomposite which is remarkably improved in the stability against various salts and cosmetic preparations and contains various bioactive ingredients, and its preparation method. The polymer-liposome nanocomposite comprises 1-50 wt% of a poly(methacrylic acid-co-n-alkyl methacrylate) random copolymer represented by the formula 1; and 50-99 wt% of a lipid dual layer which contains a useful bioactive ingredient, wherein n is 11-22; and x:y is 85:15 to 70:30 by mol. Preferably the poly(methacrylic acid-co-n-alkyl methacrylate) random copolymer has a number average molecular weight of 10,000-50,000 and a particle size of 100-400 nm. The method comprises the steps of mixing the copolymer of the formula 1, a lipid and a bioactive ingredient in an organic solvent miscible with water to dissolve them at 50-70 deg.C; mixing the obtained mixture with the heated water of 50-70 deg.C to disperse firstly with a homogenizer; and preparing nanoparticles from the mixture by using a homogenizer.

Description

Preparation of pH-responsive polymer-liposome complexes with various hydrophobic bioactive materials and the composition of skin external application containing the same }

1: Schematic diagram of a polymer-liposomal nanocomposite containing a useful bioactive component

Figure 2: Betulin crystal photo in aqueous solution through a polarizing microscope

3: Photograph of the polymer-liposomal nanocomposite (Example 1) containing betulin in aqueous solution through a polarizing microscope

Figure 4: Photo oleanolic acid crystals in aqueous solution through a polarizing microscope

FIG. 5: A photograph of a polymer-liposome nanocomposite (Example 5) containing oleanolic acid in aqueous solution through a polarizing microscope

Figure 6: Diosmethine crystals in aqueous solution through a polarizing microscope

7: Photograph of lipid-cholesterol based liposomes (Comparative Example 4) in which diosmethine in aqueous solution was effectively not incorporated through a polarizing microscope

8: Photograph of the polymer-liposomal nanocomposite (Example 7) containing diosmethine in aqueous solution through a polarizing microscope

9: Photograph showing the structural change according to the acidity of the solution in the aqueous solution of Experimental Example 3

10: Graph showing particle size distribution of nanoemulsion of Experimental Example 4

11: Graph showing the particle size distribution of the polymer-liposomal nanocomposite (Example 2) containing the betulin of Experimental Example 4

12: Graph showing particle size distribution of lipid-cholesterol-based liposomes (Comparative Example 1) containing betulin of Experimental Example 4

FIG. 13: Graph showing particle size distribution of a nanoemulsion formulation including a polymer-liposomal nanocomposite containing betulin of Experimental Example 4

14: Graph showing the particle size distribution of the nanoemulsion formulation including the lipid-cholesterol-based liposome of Experimental Example 4

The present invention relates to a polymer-liposomal nanocomposite having an acidic sensitivity incorporating an oil-soluble physiologically active ingredient, a method for preparing the same, and a cosmetic composition containing the same, and specifically, an acidic sensitive polymer composed of methacrylic acid and n-alkyl methacrylate. Useful physiology that is sensitive to acidity changes due to polymer-liposome nanocomposites or nanoemulsions prepared using a variety of useful bioactive substances, including triterpenoids similar to the structure of bilipids and cholesterol. Not only can the active ingredient be effectively released in vivo to enhance the effect as a carrier, but also can be stably present in an aqueous solution containing various salts and in cosmetic formulations and liposomes prepared based on conventional lipid-cholesterol.The present invention relates to a designed polymer-lipid based drug carrier and a cosmetic composition containing the same.

Liposomes, one of the most popular materials for gene delivery and drug delivery today, are thermodynamically stable in water-soluble inner cores and lipid vesicles, and have a lipid bilayer that surrounds at least one inner core. In liposomes, water soluble drugs are included between the water soluble channels found between successive lipid bilayers, while oil soluble drugs are located within the lipid bilayer itself. Lipid bilayers, on the other hand, may overlap several layers, such as sandwiches, and this structure is called lamellar structure. Lipid structures made of lamellar structures also associate roundly like the surface of a ball and, in the case of oil-soluble materials, are distributed in the hydrocarbon chain layer of the lipid.

Early liposomes developed as drug carriers were not used as an effective system due to colloid and biological instability, but since the stability has been improved, liposome based products of antibacterial and anticancer agents have been developed. Liposomes, by themselves, are also useful for lowering toxicity and delivering physiologically active substances in the long term by entrapping in itself a compound that exhibits utility or that is not acceptable at therapeutic doses.

In general, cholesterol is added in a proportion to lipids in the preparation of liposomes to stabilize the lipid bilayer structure of liposomes. Numerous studies have shown that the addition of cholesterol to liposomes in the preparation of liposomes helps to form structurally and thermodynamically more stable spherical liposome lipid bilayer structures. Analyzes of the rate indicate that lipid bilayers of cholesterol-containing liposomes are more stable than those of other liposomes (Diana Velluto, et al., Colloids and Surfaces B: Biointerfaces , 40, 2005, 11-18) . ). In addition, many studies have been reported to incorporate cholesterol derivatives or substances with cholesterol-like structures into the lipid bilayer of liposomes. A substance having a structure similar to cholesterol includes triterpenoids, which are naturally occurring physiologically active ingredients, and include pentacyclic compounds such as ursolic acid, oleanolic acid, and betulin. Collectively, it has been reported to have anti-cancer, hepatoprotective, anti-inflammatory, anti-ulcer, antibacterial, antitumor hemostasis, antiviral, collagen synthesis of skin, promoting skin lipid synthesis and wound healing.

However, incorporation of triterpenoids into general lipid-cholesterol-based liposomes shows stability of liposome structure in aqueous solution, but intrinsic liposome structure in cosmetic formulations in which various salts used in the biological environment or various surfactants are used. Since lipids become unstable, common lipid-cholesterol based liposomes incorporating triterpenoids have been limited in their use. In addition, in the case of other useful bioactive components such as flavonoids in addition to triterpenoids, not only the biologically active component is effectively incorporated into general lipid-cholesterol-based liposomes, but also the lipid-cholesterol containing the bioactive component in an aqueous solution even if it is incorporated. It was common for the stability of the base liposomes to be poor.

Therefore, in order for liposomes containing oil-soluble bioactive ingredients to be used in various fields, long-term stability of liposomes should be secured without crystallization of oil-soluble bioactive ingredients incorporated into liposomes in formulations containing various surfactants or in aqueous solutions containing various salts. In addition, there is an urgent need to develop a liposome system that effectively releases a high concentration of physiologically active ingredient in vivo to enhance its effect as a carrier. The need for such liposome systems is applicable to triterpenoids as well as all useful bioactive components that can be incorporated into liposomes.

In order to solve the conventional problems, the present inventors have incorporated various types of soluble bioactive components into a liposome lipid bilayer, and have an effective delivery and release effect of soluble bioactive components, and at the same time, bioactive components in an in vivo environment and cosmetic formulations. A study was conducted on liposome systems that can be stably present without crystallization of. As a result, when the polymer-liposomal nanocomposite is prepared using an acidity-sensitive polymer having a specific composition and composition, it has the same structure as the liposomes prepared based on lipid-cholesterol and exhibits sensitivity to specific changes in the acidity environment. Not only can increase the drug delivery efficiency, but also maintain long-term stability in the aqueous solution or in the presence of various salts without crystallization of physiologically active ingredients, and even in the cosmetic formulation, liposome structure without crystallization of physiologically active ingredients It has been found to be stable and the present invention has been completed.

This solved the problem of not effectively incorporating various useful bioactive ingredients into liposomes in the existing general lipid-cholesterol-based liposome system, and inducing the efficiency improvement of useful bioactive ingredients due to rapid structural collapse at specific acidity. It is meaningful that the present invention simultaneously meets the requirement to maintain particle safety without any precipitation of physiologically active ingredients under any storage condition. In addition, the present invention proves to maintain improved stability in various salts and cosmetic formulations than the lipid-cholesterol-based liposome system, which is widely known.

The present invention is a liposome of a polymer-liposomal nanocomposite prepared with acidic sensitive random copolymers and lipids composed of two monomers, methacrylic acid and n-alkyl methacrylate. It is an object of the present invention to provide a method for incorporating various types of useful bioactive components into a lipid bilayer.

Another object of the present invention has a particle size of 100 ~ 400 nm, depending on the concentration of the polymer and lipids used and the concentration of the useful bioactive component incorporated into the lipid bilayer of liposomes, the shape of the particles is a lipid containing no polymer- The present invention provides a polymer-liposomal nanocomposite that maintains the same form as a cholesterol-based liposome and has a structure in which a hydrophobic portion of the polymer is associated together between a lipid-bioactive component-based lipid bilayer.

Another object of the present invention is to effectively incorporate various kinds of useful bioactive components into the polymer-liposomal nanocomposites, which have limited effective incorporation into general lipid-cholesterol-based liposomes.

Still another object of the present invention is to move to a target organ in a state in which an oil-soluble bioactive component, which is not easily delivered in vivo due to a relatively large molecular weight, chemical structure, or poor solubility, is stably incorporated in a lipid bilayer using an acidity sensitive polymer. In order to provide a polymer-liposomal nanocomposite capable of maximizing the inherent efficacy of the active ingredient by enhancing the effect of the carrier by disrupting the lipid membrane and releasing a bioactive component in response to specific changes in the acidity environment of the target organ. have.

The present invention relates to an acidity sensitive polymer-liposomal nanocomposite containing an oil-soluble physiologically active ingredient and a method for preparing the same, wherein the polymer-liposomal nanocomposite containing an oil-soluble physiologically active ingredient is a methacrylic acid of formula 1 ) And n-alkyl methacrylate (n-alkyl methacrylate) includes an acid-sensitive polymer composed of two monomers, and the lipid-liposome nanocomposite contains a useful bioactive component in the double lipid layer.

[Formula 1]

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. A topologically sensitive polymer 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 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.

Phospholipids or nitrogenous lipids having fatty acid chains having 12 to 24 carbon atoms may be used as components of the lipid bilayer used to prepare the polymer-liposomal nanocomposites of the present invention. 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 phosphatidylethanolamine, diphosphatidylglycerol, cardiolipin, plasmogen; Hydrogenated products of natural phospholipids; Dicetylphosphate, distearoylphosphatidylcholine, dioleoylphosphatidylethanolamine, dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylethanolamine, dipalmitoylphosphatidylserine, eleostaroylphosphatidylcholine, eleostaroylolamine and Synthetic lipids of Eleo stearoylphosphatidylserine; Derivatives of synthetic lipids; And one or two or more mixtures selected from 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, 1: 1, 2: 1, 3: 1, 4: 1, 5: 1, 1 in the molar ratio of 5: 1 to 1: 5, respectively. It may be mixed in various ways, such as: 5, 1: 4, 1: 3 or 1: 2. 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 poly (methacrylic acid-co-n-alkyl methacrylate) random copolymer and the lipid bilayer used in the polymer-liposomal nanocomposite according to the present invention is poly (methacrylic acid-co-n- 1-50% by weight, preferably 5-30% by weight, of poly (methacrylic acid-co-n-alkyl methacrylate) random copolymers, based on the total weight of the alkyl methacrylate) random copolymer and the lipid bilayer And 50-99% by weight of lipids, preferably 50-90% by weight. In addition, the content of the acidity sensitive polymer and lipid used in the polymer-liposomal nanocomposite is 0.001 to 15% by weight, and preferably 1 to 10% by weight relative to the polymer-liposomal nanocomposite.

The bioactive component incorporated into the lipid bilayer of the polymer-liposomal nanocomposite of the present invention is an oil-soluble bioactive component, which has a structure similar to cholesterol, such as ursolic acid, oleanolic acid, and betulinic acid. ), Betulin, triterpenoids of b-boswellic acid, diosmetin, quercetin, flavonoids of genestein, phloretin and sperm extracts One component selected from contains 0.001 to 5% by weight, preferably 0.1 to 2% by weight based on the entire polymer-liposomal nanocomposite.

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-n-alkylmethacrylate) random copolymer, the lipid and the oil-soluble physiologically active ingredient of formula (1) with an organic solvent which can be mixed with water, and warming it to 50-70 ° C. ;

b) mixing the mixture of a) with water warmed to 50-70 ° C. and dispersing it first with an emulsifier; And

c) prepared by the method comprising the step of nanoparticulating the mixture of b) with an emulsifier.

[Formula 1]

Cholesterol may be further mixed in step a), and the organic solvent that can be mixed with water used is not limited, but is preferably selected from ethanol, methanol, isopropyl alcohol, acetone and tetrahydrofuran. In addition, the water heated in step b) may further contain a water-soluble active ingredient. Step c) of the nanoparticles is a high pressure emulsifier, the pressure can be adjusted to 100 ~ 1000 bar according to the desired particle size and dispersion degree, preferably 500 ~ 1000 bar is suitable.

The method may further include adjusting the concentration of the polymer-liposomal nanocomposite by removing the residual organic solvent and water using a rotary evaporator after step c) according to the organic solvent used.

The polymer-liposomal nanocomposite of the present invention obtained by the above production method has a particle size of 100 to 400 nm depending on the concentration of the polymer and lipid and the concentration of the useful bioactive component incorporated into the lipid bilayer of the liposome, and contains the polymer. It maintains the same particle morphology as the lipid-cholesterol-based liposomes that are not present, and as shown in FIG. 1, the hydrophobic portion of the polymer has a structure associated with the lipid-bioactive ingredient-based lipid bilayer as well as sensitive sensitivity to acidity change. It can effectively release the physiologically active ingredient incorporated into the inner box in the cell to enhance the effect as a carrier. In addition, as shown in FIG. 1, the polymer is associated with a lipid-physiologically active ingredient-based double lipid layer and simultaneously binds the double lipid layer and protects the outer wall of the liposome, thereby destabilizing various liposome structures present in the water phase. It stably maintains the structure of the liposome and helps to prevent the useful bioactive components incorporated into the double lipid layer from crystallizing in the aqueous solution.

The cosmetic composition containing the polymer-liposomal nanocomposite containing the useful bioactive component of the present invention is not particularly limited in its formulation, and it is a flexible longevity, astringent longevity, nutrient longevity, nutrition cream, massage cream, eye It can be formulated into cream, eye essence, essence, cleansing cream, cleansing lotion, cleansing foam, cleansing water, pack, powder, makeup base, foundation, body lotion, body cream, body oil, body essence, body cleanser, etc. .

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

Example 1-10 Preparation of Acidity Sensitive Polymer-Liposome Nanocomposites Containing Oil-Soluble Bioactive Components

Poly (methacrylic acid-co-stearyl methacrylate) copolymer (mol composition 70:30, number average molecular weight 26,000) for Examples 1-9, 100% hydrogenated oleyl-palmitoyl / oleyl -Stearyl phosphatidyl choline mixture (lipoid S100-3) and oil-soluble physiologically active ingredients in the weight composition ratio shown in Table 1 below by heating and dissolving in an organic solvent of 60 ℃ to prepare a mixed solution and mixed the mixture with water at 60 ℃ After mixing and stirring at a speed of 5000 rpm for 5 minutes using a homogenizer to prepare a polymer-liposome complex containing the first dispersed useful bioactive component, a high pressure homogenizer (1000 bar, 3 cycle) was used. To nanoparticles. Residual organic solvent was removed using a rotary evaporator, and a certain amount of water was also removed by distillation to control the concentration of the soluble bioactive component and the polymer-liposomal nanocomposite.

In the case of Example 10, except that a poly (methacrylic acid-co-stearyl methacrylate) copolymer (mol composition 85:15, number average molecular weight 20,000) is used, An acidity sensitive polymer-liposomal nanocomposite containing the active ingredient was prepared.

Table 1 Components and Compositions Used in Polymer-Liposomal Nanocomposites Containing Oil-Soluble Bioactive Components

[Comparative Example 1-6] Preparation of Lipid-Cholesterol-Based Liposomes Impregnated with Soluble Bioactive Components

In Comparative Example 1-6, the organic solvent of 60 ° C. in the weight composition ratio of 100% hydrogenated oleyl-palmitoyl / oleoleyl-stearyl phosphatidyl choline mixture (lipoid S100-3) and cholesterol shown in Table 1 below The solution was prepared by heating and dissolving the mixture, followed by mixing the mixture with water at 60 ° C. and mixing and stirring the mixture for 5 minutes at a speed of 5000 rpm with an homogenizer, thereby polymer-liposome complex containing the first dispersed useful bioactive component. After preparing the nanoparticles using a high pressure homogenizer (1000bar, 3cycle). Residual organic solvent was removed using a rotary evaporator, and a certain amount of water was also distilled out to adjust the concentration of the soluble bioactive component and the polymer-liposomal nanocomposite.

TABLE 2 Components and Compositions Used in Lipid-Cholesterol-Based Liposomes Impregnated with Soluble Bioactive Components

Experimental Example 1 Structural Analysis of Polymer-Liposomal Nanocomposites and Lipid-Cholesterol-Based Liposomes Containing Useful Bioactive Components

The particle size of the polymer-liposomal nanocomposite containing the useful bioactive component of Example 1-10 and the lipid-cholesterol-based liposome containing the useful bioactive component of Comparative Example 1-6 (Dynamic light scattering, The results measured using Zetasizer 3000HSa Malvern Instruments) are shown in Table 3 below.

Table 3 Results of Particle Size Measurement of Polymer-Liposome Nanocomposites and Lipid-Cholesterol-Based Liposomes

As shown in Table 3, the polymer-liposomal nanocomposite of the example in which the oil-soluble physiologically active component was incorporated had a stable form of liposomes having a constant particle size compared to the lipid-cholesterol-based liposome of the comparative example in which the oil-soluble physiologically active component was incorporated. .

The polymer-liposomal nanocomposites of Example 1-6 containing triterpenoids such as betulin, ulsolic acid, and oleanolic acid, and lipid-cholesterol-based liposomes of 1-3 were all formed stable liposomes with particle size. Unlike other useful physiologically active ingredients, triterpenoids have a cholesterol-like structure, which helps to form a structural, thermodynamically more stable spherical liposome lipid bilayer structure as shown in FIG. 1. Because of giving.

On the other hand, it was found that the particle size of the polymer-liposomal nanocomposite of the example containing the acidity sensitive polymer was larger than the particle size of the lipid-cholesterol-based liposome of the comparative example which does not contain the acidity sensitive polymer.

In addition, the polymer-liposomal nanocomposite of Example 7-10 containing useful bioactive ingredients such as diosmethin, floretine, and sperm extract except for triterpenoids has a uniform particle size to form stable liposomes. Lipid-cholesterol-based liposomes of Comparative Example 4-6 caused precipitation and did not form liposomes containing the useful bioactive components.

Therefore, it was found that the acidity sensitive polymer contained in the polymer-liposomal nanocomposite of the present invention acts as a supporter to impart acidic sensitivity to the lipid bilayer of liposomes and to stabilize the lipid bilayer. .

2-8 show the betulin crystal in aqueous solution, Example 1, the oleanolic acid crystal in aqueous solution, Example 5, the diosmethine crystal in aqueous solution, and the photographs of Comparative Examples 4 and 7 using polarizing microscopes. From 2 to 8, the polymer-liposome nanocomposites of Examples 1, 5, and 7 in which betulin, oleanolic acid, and diosmethine are present as crystals in an aqueous solution, and in which betulin, oleanolic acid, and diosmethine are incorporated, It was observed that the crystals of tulin, oleanolic acid and diosmethin were not precipitated, and also the crystals were precipitated because the liposomes based on conventional lipid-cholesterol were not effectively incorporated.

It can be seen from Figures 2 to 8 that the useful bioactive components incorporated into the polymer-liposomal nanocomposites are dispersed in the molecular unit without crystallization, through which the useful bioactive components are stable in the lipid bilayer of the polymer-liposomal nanocomposites. Was found to be.

Experimental Example 2 Evaluation of Stability of Polymer-Liposome Nanocomposites and Lipid-Cholesterol-Based Liposomes Incorporating Oil-soluble Bioactive Components

Solutions prepared to observe the long-term stability of the lipid-cholesterol-based liposomes containing the polymer-liposomal nanocomposites containing the useful bioactive components of Examples 1, 2, 7 and 8 and the soluble bioactive components of Comparative Example 1 Was stored at low temperature (2 ℃), room temperature (25 ℃) and high temperature (40 ℃) respectively to determine the change in particle size with time using a dynamic light scattering device (Dynamic light scattering, Zetasizer 3000HSa Malvern Instruments) 4 is shown.

TABLE 4

As shown in Table 4, the polymer-liposomal nanocomposites of Comparative Examples 1, 2, 7, and 8, regardless of the type of the useful bioactive component incorporated into the polymer-liposomal nanocomposite and the lipid-cholesterol-based liposome Lipid-cholesterol-based liposomes showed long-term stability in 2 ° C and 25 ° C aqueous solutions, even when stored at 2 ° C and 25 ° C.

On the other hand, the polymer-liposomal nanocomposites of Examples 1, 2, 7 and 8 and the lipid-cholesterol based liposomes of Comparative Example 1, which were stored in an aqueous solution at 40 ° C., were increased significantly compared to other particle sizes at the time of preparation. This is due to the volume expansion of liposomes, as the self-association of lipid components constituting the lipid bilayer of liposomes at a high temperature of 40 ° C is relatively loose compared to the low temperature and room temperature of 2 ° C and 25 ° C.

Salts in the lipid-cholesterol based liposomes of the polymer-liposomal nanocomposite of Example 2 and the comparative example 1 in sodium phosphate buffer and N-2-hydroxyethyl-piperazine-N´-2-ethan sulfonate (HEPES) buffer The results of observing long-term stability according to the addition of (salt) are shown in Table 5 below.

TABLE 5

As shown in Table 5, in the case of the betulin-containing polymer-liposomal nanocomposite of Example 2, the particle size and the polydispersity according to the presence or absence of salt when the salt is not present in the buffer solution and the salt is added. Is not significantly different in the long term, but in the case of the lipid-cholesterol-based liposome containing betulin of Comparative Example 1, the increase in particle size and polydispersity in sodium phosphate buffer solution containing sodium phosphate and sodium chloride (NaCl) As a result, the structure became unstable, and when stored in HEPES buffer solution, the structure collapsed and precipitated after 1 week.

Therefore, from the results of Table 4 and Table 5, the polymer-liposomal nanocomposite containing the soluble bioactive component was used in an aqueous solution containing various salts used in the biological environment than the lipid-cholesterol-based liposome containing the soluble bioactive component. It was found that the structure was much more stable.

Experimental Example 3 Observation of Acidity Sensitivity of Betulin-Containing Polymer-Liposome Nanocomposites and Lipid-Cholesterol-Based Liposomes

In order to observe the acidity sensitivity of the polymer-liposomal nanocomposite containing the oil-soluble bioactive component of the present invention, the polymer-liposomal nanocomposite of Example 2 containing the betulin as the oil-soluble bioactive component and the lipid-cholesterol base of Comparative Example 1 The liposomes were diluted with a hydrochloric acid solution at a concentration of 0.1% by weight in a sodium phosphate buffer solution titrated at 3, 4, 5, 6 and 7.4 to observe the state of the solution. , The change in particle size according to the acidity of the solution is shown in Table 6 and FIG.

9, the betulin-containing lipid-cholesterol-based liposome of Comparative Example 1 was well dispersed in the aqueous solution regardless of the acidity of the solution, but the betaline-containing polymer-liposomal nanocomposite contained the solution in the acidity of 5 or more. Although well dispersed in the liquid phase, it can be observed that in a solution having an acidity of 5 or less, no more stability is maintained and the inherent structure collapses and precipitation occurs.

On the other hand, lipid-cholesterol-based liposomes containing betulin of Comparative Example 1 appear to be well dispersed regardless of the acidity of the solution in FIG. 9, but the acidity of the solution is different from that dispersed in distilled water as shown in Table 6 below. As a result, it was observed that the particle size and polydispersity became very large.

In addition, the betulin-embedded polymer-liposomal nanocomposite of Example 2 maintained the particle shape stably at an acidity of 5 or more, but at a lower acidity, the stability of the polymer-liposomal nanocomposite was sharply lowered, resulting in rapid structural collapse. This is because the acidic sensitive polymer shows a sharp morphological change at an acidity of 5 or less, which collapses the structure of the nanocomposite.

TABLE 6

Experimental Example 4 Evaluation of Stability of Betulin-Containing Polymer-Liposome Nanocomposites and Lipid-Cholesterol-Based Liposomes in Nanoemulsion Cosmetic Formulations

To compare the structural stability in the cosmetic nanoemulsion formulation of the polymer-liposomal nanocomposites and lipid-cholesterol-based liposomes containing the useful bioactive component of the present invention, the solutions prepared in Example 2 and Comparative Example 1 10 wt% of each of the nanoemulsion formulations having an average particle size of 137 nm and formulated in the composition and content of the mixture was stirred for 5 minutes at a speed of 7000 rpm at room temperature with an homogenizer and then degassed. Nanoemulsion formulations were prepared comprising lean-containing polymer-liposomal nanocomposites and lipid-cholesterol based liposomes, respectively.

The nanoemulsion, the polymer-liposomal nanocomposite of Example 2 containing betulin and the lipid-cholesterol liposome of Comparative Example 1, the nanoemulsion formulation comprising the polymer-liposomal nanocomposite containing betulin and lipid-cholesterol-based liposome Each of the nanoemulsion formulations were stored at room temperature for 4 weeks, and then the particle size was measured using a dynamic light scattering apparatus (Dynatas light 3000 HSA Malvern Instruments), and the graphs are shown in FIGS. 10 to 14.

TABLE 7

10 is a nanoemulsion prepared in the composition of Table 7, Figure 11 is a polymer-liposomal nanocomposite containing the betulin of Example 2, Figure 12 is a lipid-cholesterol-based liposome containing the betulin of Comparative Example 1, Figure 13 A nanoemulsion formulation comprising a betulin-containing polymer-liposomal nanocomposite and FIG. 14 shows a particle size distribution of a nanoemulsion formulation comprising a betulin-containing lipid-cholesterol based liposome.

It can be seen from FIGS. 10 to 14 that the nanoemulsion formulation comprising the betulin-containing polymer-liposomal nanocomposite has much better structural stability in the formulation than the nanoemulsion formulation including the lipid-cholesterol-based liposome containing the betulin. there was. In addition, it was found that lipid-cholesterol-based liposomes containing betulin do not maintain their intrinsic liposome structure while being mixed in the nanoemulsion formulation.

As described above, the polymer-liposomal nanocomposite containing the oil-soluble physiologically active ingredient according to the present invention is introduced into an acid-sensitive polymer, so that the incorporation into the lipid-cholesterol-based liposome can be incorporated into a conventional lipid-cholesterol-based liposome. Various useful physiologically active ingredients that could not be stably incorporated into the lipid bilayer of the polymer-liposomal nanocomposite, and the structure of the polymer-liposome nanocomposite according to the change in the acidity of the aqueous solution in order to enhance the intracellular delivery ability of the bioactive ingredient. Is designed to change.

In addition, the polymer-liposomal nanocomposite according to the present invention was able to incorporate useful bioactive components other than triterpenoids, which could not be incorporated into existing general lipid-cholesterol-based liposomes, into the lipid bilayer, and is widely known lipid-cholesterol-based. It has much improved stability for various salts and cosmetic formulations than liposomes, so its value is very valuable.

Claims (9)

Polymer-liposomal nanoparticles containing a poly (methacrylic acid-co-n-alkylmethacrylate) random copolymer represented by Formula 1 below and a lipid bilayer, in which an oil-soluble bioactive component is incorporated into the lipid bilayer Complex. [Formula 1]

The method of claim 1, 1-50 weight of poly (methacrylic acid-co-n-alkylmethacrylate) random copolymer based on the total weight of the poly (methacrylic acid-co-n-alkylmethacrylate) random copolymer and the lipid bilayer Polymer-liposomal nanocomposite, characterized in that it contains% and 50-99% by weight lipid bilayer. The method of claim 1, Polymer-liposomal nanocomposite, characterized in that the number average molecular weight of the poly (methacrylic acid-co-n-alkyl methacrylate) random copolymer is 10,000 to 50,000. The method of claim 1, The lipid bilayer is yolk lecithin (phosphatidylcholine), soybean lecithin, lysolecithin, sphingomyelin, phosphatidylic acid, phosphatidylserine, phosphatidylglycerol, phosphatidyl inositol having 12 to 24 carbon chains, Natural phospholipids selected from phosphatidylethanolamine, diphosphatidylglycerol, cardiolipin, plasmogen; Hydrogenated products obtainable from natural phospholipids; Dicetylophosphate, distearoylphosphatidylcholine, dioleoyl phosphatidylethanolamine, dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylethanolamine, dipalmitoylphosphatidylserine, eleostaroyl phosphatidylcholine, eleostaroylphosphamine and eldiethanol Synthetic lipids of osteoaroyl phosphatidylserine; Derivatives of synthetic lipids; And fatty acid mixtures obtainable by hydrolysis of synthetic lipids. The polymer-liposomal nanocomposites according to claim 1, or a mixture of two or more thereof. The method of claim 1, The oil-soluble physiologically active component is a polymer-liposomal nanocomposite, characterized in that it is contained in 0.001 to 5% by weight relative to the entire polymer-liposomal nanocomposite. The method of claim 1, The oil-soluble physiologically active ingredient is a single selected from the group consisting of ulsolic acid, oleanolic acid, betulinic acid, betulin, triterpenoids of boswellic acid, diosmethin, quercetin, flavonoids of florestine, floretine and sperm extracts Polymer-liposomal nanocomposite, characterized in that the component. The method of claim 1, Polymer-liposomal nanocomposite, characterized in that the particle size of the polymer-liposomal nanocomposite is 100 ~ 400nm. a) mixing the poly (methacrylic acid-co-n-alkylmethacrylate) random copolymer, the lipid and the oil-soluble physiologically active ingredient of formula (1) with an organic solvent which can be mixed with water, and warming it to 50-70 ° C. ; b) mixing the mixture of a) with water warmed to 50-70 ° C. and dispersing it first with an emulsifier; And c) A method for producing a polymer-liposomal nanocomposite containing an oil-soluble physiologically active component, comprising the step of nanoparticulating the mixed solution of b) with an emulsifier. [Formula 1]

The method of claim 8, The method of preparing a polymer-liposomal nanocomposite using an acidity-sensitive polymer, characterized in that the organic solvent that can be mixed with water in step a) is selected from ethanol, methanol, isopropyl alcohol, acetone, tetrahydrofuran.

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