KR20200127633A - Monoolein Nano Cubic Phase Containing Polysaccharide-Disulfide Compound Complex For Cosmetic And Manufacturing Method Thereof - Google Patents

Abstract

Provided in the present invention is a monoolein nanostructure for a cosmetic material for delivery of effective ingredients to the skin and the body, which is a monoolein cubic phase containing a polysaccharide-disulfide compound complex inside a water channel. The polysaccharide-disulfide compound complex is bonded to steryl trimethyl ammonium chloride inserted in a monoolein bilayer and fixed to the water channel, and thus there is an advantage of enabling a stable fixation even without modification of polysaccharide or disulfide compound. In addition, the polysaccharide-disulfide compound complex has an advantage of safely storing physiologically active substances without diffusion through a three-dimensional network formed by an electrostatic mutual bond with physiologically active substances and a disulfide bond of disulfide compounds in an external environment, and also has a feature of removing the electrostatic mutual bond and the disulfide bond to diffuse and release the physiologically active substances in an internal environment. According to the present invention, the monoolein nanostructure for a cosmetic material for delivery of effective ingredients to the skin and the body is composed of a lipid bilayer, so as to provide an excellent transdermal absorption performance, and also provide a feature of controlling and releasing the physiologically active substances only when placed in an internal environment while being absorbed into the epidermal layer without releasing the physiologically active substances when applied to the epidermal layer. Thus, the monoolein nanostructure for a cosmetic material for delivery of effective ingredients to the skin and the body of the present invention provides an advantage of being used as a functional cosmetic drug delivery system capable of controlling and releasing the physiologically active substances by actively responding to the internal environment without any artificial stimulation.

Description

Monoolein Nano Cubic Phase Containing Polysaccharide-Disulfide Compound Complex For Cosmetic And Manufacturing Method Thereof}

The present invention relates to a monoolein nanocubic phase for a cosmetic material comprising a polysaccharide-disulfide compound complex.

The monoolein cubic phase has excellent physicochemical properties and is thus attracting much attention as a drug delivery system. Monoolein may be composed of one molecule of oleic acid and one molecule of glycerol, is amphiphilic, has a packing parameter of a little greater than 1, and exists in a cubic phase in a water environment. The monoolein bilayer has a diameter of 3 to 5 nm and is surrounded by intercrossing water channels.

In the monoolein cubic phase, a material capable of sensing stimulation may be installed in a double layer or a water channel. Then, the monoolein cubic phase releases the material mounted therein in response to a specific stimulus. For example, when a heat-sensitive polymer is included in the water channel, the monoolein cubic phase releases the drug mounted therein by heat. The thermally controlled release is induced when the heat-sensitive polymer changes the structure of the water channel due to heat, and the diffusion rate of the drug loaded in the water channel is changed, thereby improving the release rate. As another example, when an ionizable polymer is included in the cubic phase, the loaded drug may be released through a water channel according to pH-dependent properties. The release of the drug is controlled not only by the structural change of the polymer according to the pH change, but also by the change in the electrostatic interaction between the polymer and the drug according to the pH change. As another example, when the light crosslinking polymer is included in the water channel, the release of the loaded drug can be controlled by irradiating light onto the cubic. When photo-thermal nanoparticles such as gold nanospheres are included in a cubic double layer, the release of drugs can be controlled by irradiating near-infrared rays. In particular, since photothermal energy causes a cubic to hexagonal transition, it is known that more efficient drug release is possible.

However, in the case of cosmetics or drug delivery systems applied to the skin and injected into the body, there is a limitation in controlling and releasing the physiologically active substances contained therein by the above artificial stimulation. Therefore, if a cosmetic material and drug delivery system that actively releases physiologically active substances in response to the internal environment after maintaining the properties of storing physiologically active substances outside the body are developed, functional cosmetics using physiologically active substances It is expected that the development will become more active.

The patent documents and references mentioned in this specification are incorporated herein by reference to the same extent as if each document was individually and clearly specified by reference.

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DOI: 10.1016/0009-3084(80)90026-2. Sagalowicz, L.; Leser, M.; Watzke, H.; Michel, M. Monoglyceride Self-assembly Structures as Delivery Vehicles. Trends Food Sci. Technol. 2006, 17,204-214. DOI: 10.1016/j.tifs.2005.12.012. Caboi, F.; Amico, G. S.; Pitzalis, P.; Monduzzi, M.; Nylander, T.; Larsson, K. Addition of Hydrophilic and Lipophilic Compounds of Biological Relevance to the Monoolein/water System. I. Phase Behavior. Chem. Phys. Lipids. 2001, 109, 47-62. DOI: 10.1016/S0009-3084(00)00200-0. Lindblom, G.; Larsson, K.; Johansson, L.; Fontell, K.; Forsen, S. The Cubic Phase of Monoglyceride-water Systems. Arguments for a Structure Based Upon Lamellar Bilayer Units. J. Am. Chem. Soc. 1979, 101, 5465-5470. DOI: 10.1021/ja00513a002. Lindblom, G.; Rilfors, L. Cubic Phases and Isotropic Structures Formed by Membrane Lipids possible Biological Relevance. Biochim. Biophys. Acta, Rev. Biomembr. 1989, 988, 221-256. DOI: 10.1016/0304-4157(89)90020-8. Ye, Z.-G.; Schmid, H. Optical, Dielectric and Polarization Studies of the Electric Field-induced Phase Transition in Pb(Mg1/3Nb2/3) O3 [PMN]. Ferroelectrics 1993, 145,83-108. DOI: 10.1080/00150199308222438. Alexandridis, P.; Olsson, U.; Lindman, B. A Record Nine Different Phases (Four Cubic, Two Hexagonal, and One Lamellar Lyotropic Liquid Crystalline and Two Micellar Solutions) in a Ternary Isothermal System of an Amphiphilic Block Copolymer and Selective Solvents (Water and Oil). Langmuir 1998, 14, 2627-2638. DOI: 10.1021/la971117c. Costa, P.; Sousa Lobo, J. M. Modeling and Comparison of Dissolution Profiles. Eur. J. Pharm. Sci. 2001, 13, 123-133. DOI: 10.1016/S0928-0987(01)00095-1. Angelov, B.; Angelova, A.; Ollivon, M.; Bourgaux, C.; Campitelli, A. Diamond-type Lipid Cubic Phase with Large Water Channels. J. Am. Chem. Soc. 2003, 125, 7188-7189. DOI: 10.1021/ja034578v. Taylor, J. M.; Cohen, S.; Mitchell, W. M. Epidermal Growth Factor: High and Low Molecular Weight Forms. Proc. Natl. Acad. Sci. 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The present invention relates to a monoolein nanocubic phase for a cosmetic material comprising a polysaccharide-disulfide compound complex for delivery of active ingredients to the skin and body, and a physiologically active substance is safely stored inside the structure outside the body before application to the skin. It relates to a drug delivery system for a cosmetic material that is absorbed into the skin and introduced into the body to increase the diffusion of the drug and released.

Accordingly, an object of the present invention is to provide a monooleion nanostructure for a cosmetic material including a polysaccharide-disulfide compound complex and a method of manufacturing the same for the delivery of active ingredients to the skin and body.

Other objects and technical features of the present invention are presented in more detail by the following detailed description, claims, and drawings.

The present invention provides a monooleion nanostructure for a cosmetic material for delivery of active ingredients to the skin and body, including a polysaccharide-disulfide compound complex.

The monoolein nanostructure is a monoolein cubic phase, and steryl trimethyl ammonium chloride is inserted into a double layer.

The polysaccharide-disulfide compound complex is fixed to the bilayer constituting the water channel of the monooleion nanostructure for drug delivery in the body, and the isoelectric point inside the polysaccharide-disulfide compound complex has a physiological activity of pH 4 to 6 It further contains substances.

In particular, when the physiologically active substance is present in a solution having a pH of 7 to 8 and a concentration of reduced glutathione of 1 to 10 mM, the monooleion nanostructure for drug delivery in the body, from the inside of the polysaccharide-disulfide compound complex It diffuses into the water channel and has a feature of improving the emission rate.

The present invention is a first step of preparing a mixed melt by mixing monoolein and steryl trimethyl ammonium chloride at 45 to 55 ℃ and cooling to 30 to 40 ℃; The second step of preparing a monooleion nanostructure by adding a polysccharide, a disulfide compound, and a physiologically active substance while maintaining the mixed melt at 30 to 40°C and maintaining it at 20 to 25°C. It provides a method of manufacturing a monooleion nanostructure for a cosmetic material for delivery of active ingredients to the skin and body, including.

The monoolein and steryl trimethyl ammonium chloride are mixed in a molar ratio of 1:4 to 1:6, and the polysccharide and disulfide compound are the carboxyl group of the polysaccharide and the amino group of the disulfide compound is 1:0.6 To 1:1 molar ratio.

The monooleion nanostructure for a cosmetic material for delivery of active ingredients to the skin and body of the present invention is a monoolein cubic phase containing a polysaccharide-disulfide compound complex inside a water channel.

Since the polysaccharide-disulfide compound complex is bonded to the steryl trimethyl ammonium chloride inserted in the monoolein bilayer and fixed to the water channel, there is an advantage that stable fixation is possible without modification of the polysaccharide or disulfide compound.

In addition, the polysaccharide-disulfide compound complex has the advantage of being able to safely store physiologically active substances without diffusion through a three-dimensional network formed by electrostatic interactions with bioactive substances and disulfide bonds of disulfide compounds in an external environment. Electrostatic mutual bonds and disulfide bonds are removed so that the bioactive material is diffused and released.

The monooleion nanostructure for a cosmetic material for delivery of active ingredients to the skin and body of the present invention is composed of a lipid bilayer and has excellent transdermal absorption performance and does not release physiologically active substances when applied to the epidermal layer. Bioactive substances are controlled and released only when absorbed into the epidermal layer and placed in the internal environment.

Therefore, the monoolein nanostructure for cosmetic materials for delivery of active ingredients to the skin and body of the present invention can be used as a functional cosmetic drug delivery system capable of controlling and releasing physiologically active substances by actively responding to the body environment without any artificial stimulation. There is this.

1 shows a schematic diagram of a monoolein nanostructure for drug delivery in the body of the present invention. Panel (A) shows the state in which the pH is below the isoelectric point and the diffusion of physiologically active substances is suppressed in the non-reducing state, and Panel (B) shows the state in which the pH is above the isoelectric point and the diffusion of the biologically active substance is promoted in the reducing state, thereby increasing the amount of release. Show the state.
2 shows the light scattering intensity measurement result of the present invention. The closed circle in panel (A) shows the degree of change in light scattering intensity according to the pH change, and the open circle shows the degree of change in the light scattering intensity when the molar ratio of carboxyl group and amino group is changed. Panel (B) shows the change in the intensity of light scattering related to the generation of the alginate-epithelial cell growth factor conjugate of the present invention according to the pH change.
3 shows an electron microscope analysis result of the monoolein nanostructure for drug delivery in the body of the present invention. Panel (A) relates to a monoolein cubic phase (0/0), panel (B) relates to a monoolein cubic phase (1/0), and panel (C) relates to a monoolein cubic phase (1). /0.8), and panel (D) relates to a monoolein cubic phase (1/0.8/EGF).
Figure 4 shows the results of differential scanning calorimeter analysis of the monoolein nanostructure for drug delivery in the body of the present invention. In panel (A), a is for a monoolein cubic phase (0/0), b is for a monoolein cubic phase (1/0), and c is for a monoolein cubic phase (1/0.5). And d is for the monoolein cubic phase (1/0.8). In panel (B), a is for a monoolein cubic phase (1/0/EGF), b is for a monoolein cubic phase (1/0.5/EGF), and c is for a monoolein cubic phase (1 /0.8/EGF).
FIG. 5 shows the results of polarization microscopy analysis of the monoolein nanostructure for drug delivery in the body of the present invention. Panel (A) relates to a monoolein cubic phase (0/0), panel (B) relates to a monoolein cubic phase (1/0), and panel (C) relates to a monoolein cubic phase (1). /0.5), and panel (D) relates to a monoolein cubic phase (1/0.8).
Figure 6 is for the analysis of the release rate of the bioactive material for the monoolein nanostructure for drug delivery in the body of the present invention. Panel (A) shows the result of the release buffer at pH 4.0, the closed circle represents the case of reduced glutathione 0mM, the open circle represents the reduced glutathione 2mM, and the closed triangle represents the case of reduced glutathione 10mM. Panel (B) shows the result of the release buffer at pH 5.5, the closed circle represents the case of reduced glutathione 0mM, the open circle represents the reduced glutathione 2mM, and the closed triangle represents the case of reduced glutathione 10mM. Panel (C) shows the result of the release buffer of pH 7.4, the closed circle represents the case of reduced glutathione 0mM, the open circle represents the case of reduced glutathione 2mM, and the closed triangle represents the case of reduced glutathione 10mM.

The present invention provides a monooleion nanostructure for a cosmetic material for delivery of active ingredients to the skin and body, including a polysaccharide-disulfide compound complex.

The monoolein nanostructure is characterized by having a structure of a monoolein cubic phase, and includes a polysaccharide-disulfide compound complex capable of loading a water-soluble physiologically active substance inside the cubic phase. In particular, the monoolein nanostructure for a cosmetic material for delivery of active ingredients to the skin and the body of the present invention has the effect of controlling and releasing the physiologically active substances mounted therein in response to the pH and reducing environment in the body.

The monoolein is glycerol monooleate having a packing parameter of slightly greater than 1. The mono-olein may form a cubic phase having an isotropic structure in an aqueous phase, and the cubic phase may be optically transparent and have a water channel crossing the interior of the lipid matrix.

The water channel is separated by a monoolein double layer and can freely communicate with the surrounding environment as an aquatic environment. Accordingly, various kinds of water-soluble compounds, that is, drugs or bioactive substances, may be mounted inside the water channel.

The monoolein bilayer of the monoolein nanostructure for a cosmetic material for delivery of active ingredients to the skin and body of the present invention is characterized in that steryl trimethyl ammonium chloride having low cytotoxicity is inserted.

The steryl trimethyl ammonium chloride includes a positively charged head and a hydrophobic tail. When mixed with the monoolein, the hydrophobic tail is inserted into the monoolein bilayer and the head is located on the surface of the bilayer.

Particularly, the head part having a positive charge and located on the surface of the monoolein bilayer constitutes the polysaccharide-disulfide compound complex and undergoes electrostatic interaction with a polysaccharide having a negative charge. The mutual bond serves to fix the polysaccharide-disulfide compound complex on the monoolein bilayer, which becomes an important mechanism for the polysaccharide-disulfide compound complex to be located inside the water channel.

The polysaccharide-disulfide compound complex is composed of a polysaccharide containing at least one carboxyl group and a disulfide compound having at least one amino group. The polysaccharide is a polymeric carbohydrate molecule composed of a long chain of monosaccharide units formed by glycoside bonds.

The polysaccharide of the present invention is a carboxypolysaccharide (CPS) having one or more carboxyl groups. The carboxyl polysaccharide of the present invention is from the group consisting of alginate, carboxymethyl cellulose, carboxyethyl cellulose, chitin, hyaluronic acid, starch, glycogen, pectin, carboxymethyl dextran, carboxymethyl chitosan, and glycosaminoglycan, and heparin. It may be any one selected or a mixture of two or more, preferably alginate.

The alginate is composed of residues of mannuronate and gluronate containing a carboxy group and electrostatically interacts with the steryl trimethyl ammonium chloride inserted in the double layer, so the polysaccharide-disulfide compound It serves to fix the complex to the monoolein cubic water channel.

The disulfide compound refers to a compound capable of forming a difulfide bond in a non-reducing state and containing one or more amino groups. Preferably, the disulfide compound of the present invention is cystamine. The cystamine is a disulfide compound containing two amino groups and performs electrostatic interaction with the residue of the alginate to form a polysaccharide-disulfide compound complex and forms a disulfide bond between cystamine. The polysaccharide-disulfide compound complex forms a three-dimensional network.

In the polysaccharide-disulfide compound complex of the present invention, the polysaccharide and the disulfide compound are mixed so that the carboxyl group of the polysaccharide and the amino group of the disulfide compound have a molar ratio of 1:0.6 to 1:1. Since the polysaccharide-disulfide compound complex exists in a gel state in which a three-dimensional network is formed by disulfide bonds between the disulfide compounds in the monoolein cubic phase, the water channel of the monoolein cubic phase is closed. Therefore, the physiologically active material mounted inside the cubic phase is safely stored without diffusion to the outside.

A physiologically active substance is mounted inside the three-dimensional network of the polysaccharide-disulfide compound complex. The bioactive material does not easily diffuse into the water channel on a cubic monoolein due to the physical barrier of the three-dimensional network structure. On the contrary, when the disulfide bond is removed by reduction and the three-dimensional network of the polysaccharide-disulfide compound complex is destroyed, diffusion of the bioactive material into the water channel is promoted.

Therefore, if the molar ratio is less than 1:0.6, the number of disulfide bonds is insufficient and the three-dimensional network is not densely constructed, so that the effect of blocking the diffusion of the physiologically active material in the water channel may be insignificant. Too much, it is difficult to remove the 3D network due to the reduction reaction, so it is difficult to control the diffusion of bioactive substances.

The physiologically active substance mounted inside the three-dimensional network of the polysaccharide-disulfide compound complex is a physiologically active substance having an isoelectric point of 4 to 6.

The physiologically active substance may be a water-soluble monomolecular compound, water-soluble peptide, or water-soluble protein having physiological activity. If the pH is lower based on the isoelectric point, the surface charge is positive, and if the pH is higher based on the isoelectric point, the surface charge is present as a negative charge. It is a substance. Preferably, the physiologically active substance of the present invention is an epidermal growth factor (EGF).

The polysaccharide contained in the polysaccharide-disulfide compound complex of the present invention has a negative charge. Therefore, when the physiologically active substance is present in an environment with a pH of less than 4, it has a positive charge and electrostatically interacts with the polysaccharide. On the contrary, when the physiologically active substance of the present invention is present in an environment of pH 6 or higher, it has a negative charge and electrostatically repels the polysaccharide. Therefore, when the pH around the physiologically active substance is adjusted based on the isoelectric point, the binding of the physiologically active substance and the polysaccharide can be controlled, so that the degree of diffusion of the physiologically active substance into the water channel can be controlled.

In summary, the degree of diffusion into the water channel is determined according to the degree of formation of a three-dimensional network by the disulfide bond of the polysaccharide-disulfide compound complex and the surface charge of the bioactive substance in the bioactive substance of the present invention. It will also increase.

The monoolein nanostructure for a cosmetic material for delivery of active ingredients to the skin and body of the present invention on which the physiologically active substance of the present invention is mounted is stored in a non-reducing buffer solution having a pH of about 3 to 4. The structure present in the storage solution solidly maintains the three-dimensional network of the polysaccharide-disulfide compound complex, and the physiologically active substance mounted therein has a positive charge, thereby forming an electrostatic interaction with the polysaccharide-disulfide compound complex. External release of active substances is blocked.

On the contrary, when the monoolein nanostructure for a cosmetic material for delivery of active ingredients to the skin and body of the present invention is used as a cosmetic composition and applied to the skin, the structure is absorbed into the skin and penetrates into the body. The structure injected into the body maintains a reducing environment by a certain concentration of reduced glutathione, and is located in the body where the pH is always maintained at 7 to 8, and the disulfide bond is reduced and removed, thus destroying the three-dimensional network of the polysaccharide-disulfide compound complex. Then, the surface charge of the physiologically active material is converted into a negative charge, thereby removing electrostatic interaction with the polysaccharide. Accordingly, the physiologically active material increases in diffusion into the water channel inside the structure, and is eventually released from the nanostructure into the body.

The essential function of the drug delivery system is to safely store the drug until it is taken, then enter the body after taking it and release the drug. As described above, it is determined that the monoolein nanostructure for a cosmetic material for delivery of active ingredients to the skin and body of the present invention meets the essential functions of the drug delivery system. The monoolein nanostructure for a cosmetic material for delivery of active ingredients to the skin and body of the present invention safely stores the physiologically active substance in a pH environment below the isoelectric point of the physiologically active substance and in a non-reducing environment. In body fluids or cells corresponding to the above pH environment and reducing environment, the physiologically active substance is released to the outside of the structure.

The present invention provides a method of manufacturing a monoolein nanostructure for a cosmetic material for delivery of active ingredients to skin and body, including the following steps.

Step 1: Monoolein and steryl trimethyl ammonium chloride are mixed at 45 to 55°C and cooled to 30 to 40°C to prepare a mixed melt.

Step 2: Maintaining the mixed melt at 30 to 40°C, adding polysccharide, disulfide compound, and physiologically active material, and maintaining it at 20 to 25°C to prepare a monooleion nanostructure Step to do.

If the mixing temperature is less than 45°C, the dispersibility of the lipid molecule is lowered, resulting in a problem in the formation of the lipid bilayer, and even if the temperature is 55°C or higher, the dispersibility is not significantly improved. Preferably, the mixing is carried out at 50°C.

Since polysaccharides, disulfide compounds, and physiologically active substances are added to the mixed melt and mixed, polysaccharides, disulfide compounds, and physiologically active substances are mounted inside the water channel. For this purpose, it is preferable to maintain the temperature of the mixed melt at 30 to 40°C. When the temperature of the mixed melt is less than 30°C or exceeds 40°C, the degree of internal loading of polysaccharides, disulfide compounds and physiologically active substances decreases. Preferably, the temperature of the mixed melt is 35°C.

The present invention will be described in more detail through examples below.

Example

1. Experimental materials and methods

1.1 Experimental material

Alginate is composed of mannuronate and gluronate in a molar ratio of 69:31, molecular weight (MW) is 4000 to 7000 Da, viscosity (viscosity) is 1% An aqueous solution of 4 to 12 cP at 25°C was used. The alginate, cystamine dihydrochloride, pluronic F127, glutathione and phosphotungstic acid were purchased from Sigma Aldrich Co., MO, USA. Cell growth factors were purchased from BIO-FD&C (Incheon, South Korea).

Monoolein Monomuls 90-O 18 was provided by BASF-Korea (Seoul, South Korea), and steryl trimethyl ammonium chloride (STAC) was obtained from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). All other materials used materials of analytical level.

1.2 Light scattering analysis for alginate complexes

First, a gel-like alginate-cystamine complex was prepared and light scattering analysis was performed on it. For this, a 1% (w/v) alginate solution was prepared using a buffer solution having a pH of 3 to 9. The buffer solution was a glycine buffer solution (30mM glycine) with pH 3.0, 4.0, 9.0, MES buffer solution (30mM glycine) with pH 5.0 and 6.0, and HEPES buffer solution (30mM glycine) with pH 7.0 and 8.0. A 1% (w/v) cytamine solution was prepared using the buffer solution. In a 5 ml test tube, 2 ml of alginate solution and 1.24 ml of cystamine solution are mixed so that the concentration of alginate and cystamine is 1% (w/v), and the carboxyl group and amino group are in a 1:1 molar ratio. An alginate-cystamine complex in the state was prepared.

Light scattering of the alginate-cystamine complex was measured using a dynamic light scattering meter (Plus 90, Brookheaven instrument, USA) and measured at a 90° angle with a wavelength of 670 nm. 1% alginate solution (pH 3.0) and 1% cystamine solution (pH 3.0) were mixed in various volume ratios to confirm the formation of the complex according to the volume ratio of alginate and cystamine, so that the molar ratio of carboxyl group and amino group was 1 : 0.1 to 5, but the total amount of the alginate and cystamine mixture was kept at 1% (w/v).

Next, an alginate-epithelial cell growth factor complex was prepared and light scattering analysis was performed on it. The alginate solution (pH 3.0-9.0) was mixed with the epithelial cell growth factor solution prepared with the same buffer solution used in the preparation of the alginate solution, and the carboxyl group of the alginate and the amino group of the epithelial cell growth factor were made to have a molar ratio of 1:1. . In addition, the concentration of alginate and epithelial cell growth factor was set to 0.02% (w/v).

1.3 Preparation of monoolein cubic phase

A monoolein cubic phase containing alginate, cystamine, and epithelial cell growth factor and containing steryl trimethyl ammonium chloride in the bilayer was prepared.

First, 2 g of monoolein and 4 mg of steryl trimethyl ammonium chloride were put together in a 10 ml container, and then immersed in a constant temperature water bath maintained at 50° C. to obtain a mixture melt and then cooled to 35° C.

Next, cystamine (0 mg, 6.2 mg, or 9.9 mg), alginate (0 mg, or 20 mg), and epidermal growth factor (0 mg, or 20 mg) were dissolved in distilled water with a pH of 8 and mixed. A solution was prepared, and the mixed solution was adjusted so that the molar ratio of the carboxyl group of alginate and the amino group of cystamine was 0:0, 1:0, 1:0.5, or 1:0.8, respectively.

Finally, the mixed solution was heated to 35° C. to maintain the same temperature as the mixed melt, and 0.857 ml of the heated mixed solution was slowly added to the mixed melt. After that, the cap was covered and wrapped with aluminum foil, and the mixed melt was completely hydrated at room temperature (20 to 23°C) to prepare a transparent gel-like monoolein cubic phase.

The prepared monoolein cubic phase was represented by a monoolein cubic phase (a/b/c), respectively. Where a means the number of moles of alginate, b means the number of moles of cystamine, c means epithelial growth factor, but when EGF is indicated, it means that 20 mg of epithelial growth factor is used, and when indicated as 0 It means using 0 mg of epithelial cell growth factor. In the case of epithelial cell growth factor, it was included in the cubic phase and was added after calculating the molar ratio.

The monoolein cubic phase prepared by the above method is shown in Table 1 below.

Monoolein STAC Alginate Cystamine EGF Example 1 Monoolein Cubic Prize (0/0) 2g 4mg 0mg 0mg 0mg Example 2 Monoolein Cubic Prize (1/0) 2g 4mg 20mg 0mg 0mg Example 3 Monoolein Cubic Prize (1/0.5) 2g 4mg 20mg 6.2mg 0mg Example 4 Monoolein Cubic Award (1/0.8) 2g 4mg 20mg 9.9mg 0mg Example 5 Monoolein cubic phase (0/0/EGF) 2g 4mg 0mg 0mg 20mg Example 6 Monoolein cubic phase (1/0/EGF) 2g 4mg 20mg 0mg 20mg Example 7 Monoolein cubic phase (1/0.5/EGF) 2g 4mg 20mg 6.2mg 20mg Example 8 Monoolein cubic phase (1/0.8/EGF) 2g 4mg 20mg 9.9mg 20mg

1.4 Analysis of monoolein cubic phase by electron microscope

The monoolein cubic phase was added 100 mg each to a 20 mL container in which 10 mL of Pluronic F127 solution (1.2% (w/v)) was dispensed, and at 30 second intervals in a water bath ultrasonicator (VC 505, Sonic & Materials, USA). It was homogenized for 30 minutes while applying a pulse. 1 ml of the cubic suspension solution micronized by the above method was mixed with 1 ml of a PTA solution (2% (w/v)), and the mixture was left at room temperature for 4 hours. The formvar/copper coated grid was wetted with the suspension of the mixture and dried in air. The sample was photographed using an electron microscope (LEO 912AB OMEGA, LEO Elektrone-mikroskopie, Germany).

1.5 Differential Scanning Calorimeter and Polarization Microscopy Analysis of Monoolein Cubic Phase

A differential scanning calorimetry (DSC) and a polarizing microscope were used to observe the phase shift induced by heat in the prepared monoolein cubic phase. After putting 5 to 7 mg of cubic phase in a sealed pan, the lid was opened and pressure was applied to seal it (Tzero Sample Press Kit, TA Instruments, USA). The pan was placed in a differential scanning calorimeter (DSC Q2000, TA Instruments, USA; installed in the Central Laboratory of Kangwon National University) and scanned at 35 to 70°C.

A monoolein cubic cell was prepared for polarization microscope analysis. First, an O-ring made of polypropylene was attached to the cover glass using a glue. Then, a sufficient amount of the monoolein cubic phase was placed in the space between the films, covered with another cover glass, and pressure was applied with a finger to make it completely close. After placing the prepared cell in a temperature control device (HS81, Mettler Toledo, USA), while heating to the same temperature range, a monoolein cubic image was photographed through a polarizing microscope.

1.6 Measurement of the amount of epithelial growth factor released

A pH 4.0, 5.5, or 7.4 buffer solution (150mM) in which reduced glutathione (GSH) is dissolved in a concentration of 0mM, 2mM, or 10mM was used as the epithelial cell growth factor release measurement solution (release solution).

After putting 3 ml of the release medium in a 10 ml container, 2 g of cubic phase (0/0/EGF), cubic phase (1/0/EGF), or cubic phase (1/0.8/EGF) were added, and I closed the lid. After 72 hours, 150 µl of the release solution was taken from the sample, and the same release solution was filled. Coomassie plus assay (Pierce Biotechnology, USA) was performed on the collected release solution to measure the concentration of epithelial cell growth factors. The degree of release of the epithelial cell growth factor refers to the amount of the epithelial cell growth factor released compared to the amount of the initial epithelial cell growth factor mounted on the cubic phase.

2. Experiment result

2.1 Light scattering analysis result of alginate complex

Panel (A) of FIG. 2 shows the degree of change in light scattering intensity of the mixed solution of the alginate-cystamine complex according to the pH and the molar ratio of the carboxyl group and the amino group.

It was confirmed that the light scattering intensity of the mixed solution was stronger as the pH was lowered. When the pH of the mixed solution was changed from 9.0 to 3.0, it was confirmed that the light scattering intensity increased from 105 kcps (kilo count per second) to 260 kcp (refer to the black dot graph in panel (A) of FIG. 2). Increasing the intensity of light scattering means that the amount of insoluble matter capable of scattering light is increased.

One molecule of cystamine has two terminal amino groups. Therefore, the amino group can cross-link with the alginate chain by electrostatic mutual bonding, which leads to the formation of an insoluble complex. In a low pH environment, the amino group has a stronger tendency to bind with hydrogen, so that the formation of electrostatic complexes is more advantageous. As a result of specifying the light scattering intensity by fixing the pH of the mixed solution to 3.0 and changing the molar ratio of the carboxyl group and the amino group, it was confirmed that the light scattering intensity was about 106 kcps when the molar ratio was 1:0 (Fig. 2 ), see white point graph). The above result means that alginate is present as an insoluble substance in the mixed solution of pH 3.0.

Since the carboxyl group exists in a non-ionized form in an acidic environment, the electrostatic repulsive force between molecules becomes relatively weak. Therefore, in the acidic environment, alginate chains are entangled with each other to exist insoluble. The light scattering result in the pH 3.0 environment is considered to be due to the entangled alginate chain.

When the molar ratio of the carboxyl group and the amino group is 1:0 to 0.5, it is confirmed that the light scattering intensity is almost constant at about 100 kcp (refer to the white dot graph in panel (A) of FIG. 2). In an acidic environment, the amino group of cystamine also has a strong tendency to bind with hydrogen.This property can be used for cross-linking with the alginate chain, and when the cross-linking is achieved, an insoluble gel-like alginate-cystamine complex is formed. do. However, according to the results of FIG. 2, it is confirmed that the light scattering value did not increase even though the molar ratio of the carboxyl group and the amino group increased to 1:0.5. This is believed to be due to the fact that the amount of cystamine was small, so that it was not possible to form an insoluble complex with alginate. In addition, when the molar ratio of carboxyl group and amino group increases from 1:0.5 to 1:1.5, it is confirmed that the light scattering intensity increases rapidly from 103 kcps to 294 kcps. This is believed to be because the amount of molecules participating in the cross-linking also increased as the relative amount of cystamine increased, resulting in the formation of an insoluble alginate-cystamine complex, which improved the intensity of light scattering.

On the other hand, when the molar ratio of the carboxyl group and the amino group is increased from 1:1.5 to 1:5, it is confirmed that the light scattering intensity rather decreases from 294 kcps to 97 kcps. The case where the light scattering intensity has a maximum value is when the electrostatic interconnection reaches the maximum value. Therefore, when the molar ratio of the carboxyl group and the amino group is 1:5, it is judged that the light scattering intensity is rather reduced because the carboxyl group of the alginate is saturated by the amino group of cystamine (refer to the white dot graph in Fig. 2). When the molar ratio of the carboxyl group of alginate and the amino group of cystamine increases from 1:1.5 to 1:5, the amino group of cystamine is stoichiometrically exceeded, thus saturating the carboxyl group of alginate. Accordingly, as the molar ratio increases from 1:1.5 to 1:5, the amount of alginate decreases, which decreases the formation of the alginate-cystamine complex, and thus the intensity of light scattering decreases.

When no alginate is used and the molar ratio of carboxyl group and amino group is 0:1, the light scattering intensity is confirmed to be close to 0kcps. This is believed to be because cystamine-alginate complex gel was not produced at all.

Panel (B) of Figure 2 shows the light scattering intensity measurement result of alginate and epithelial cell growth factor mixture according to the pH change. When the pH rises from 2.4 to 4.2, it is confirmed that the value of the light scattering intensity rises from 324 kcps to 904 kcps.

The isoelectric point of epithelial cell growth factor is pH 4.6. The epithelial cell growth factor present at the pH has a positive charge. The positively charged epithelial cell growth factor binds to a polysaccharide having a negative charge, such as alginate, which leads to the formation of complex coacervate. It is determined that the complex coacervate is the cause of the light scattering. Since the degree of ionization of the carboxyl group contained in alginate is proportional to the increase of the pH value, as long as the epithelial cell growth factor is present in a positively charged state, the amount of complex coacervate formed increases in a higher pH environment. The above result explains the result that the intensity of light scattering increases as the pH increases from 2.4 to 4.3.

When the pH rises from 4.2 to 5.0, it is confirmed that the light scattering intensity decreases from 904 kcps to 780 kcps. Since the isoelectric point of the epithelial cell growth factor is pH 4.6, when the pH is increased from 4.2 to 5.0, the charge of the epithelial cell growth factor changes from a positive charge to a negative charge. This change causes the formation of the composite coacervate to be lowered.

Also, when the pH is increased from 5.0 to 6.3, the light scattering intensity decreases from 780 kcps to 103 kcps. In the above pH range, as the pH value increases, the amount of negative charge of the epithelial cell growth factor increases. As a result, the electrostatic interaction between the positively charged alginate and the epithelial cell growth factor is reduced, and the formation of the complex coacervate is rapidly reduced, and thus the intensity of light scattering is also rapidly decreased. When the pH value rises from 6.3 to 8.0, the light scattering intensity gradually decreases from 103 kcps to 8 kcps, and when the pH value is 8.0 to 9.0, the light scattering intensity approaches "0". Epithelial cell growth factor is difficult to electrostatically bind with alginate in an environment of pH 8.0 to 9.0, but only a small amount of complex coacervate is formed. The reason is that the pH range is far from the isoelectric point of the epithelial cell growth factor and thus has a strong negative charge.

2.2 Results of electron microscopy analysis of monoolein cubic phase

Figure 3 is for monoolein cubic phase (0/0), monoolein cubic phase (1/0), monoolein cubic phase (1/0.8) and monoolein cubic phase (1/0.8/EGF) Shows the result of observation under an electron microscope. The white line means the water channel and the black line means the fat bilayer. According to the result of FIG. 3, the water channel (white line) and the fat double layer (black line) are confirmed to be bent to each other. PTA as a heavy metal component used in electron microscope sample preparation is water-soluble. For reference, during the dyeing process for sample preparation, the PTA can easily penetrate into the water channel. As a result, the electron beam does not penetrate well into the water channel, so that the water channel is observed as white. The crooked white lines and black lines are confirmed by electron microscope observations of all cubic phases, which means that all cubic phases have the same structure.

Through the above results, it was confirmed that alginate, cystamine and epithelial cell growth factor did not affect the cubic structure. The alginate, cystamine, and epithelial cell growth factor are water-soluble, so they exist only inside the water channel and there is no fear of mutual bonding with the bilayer. Therefore, it is believed that the alginate, cystamine, and epithelial cell growth factor can be safely applied without concern about the structural effect of the cubic phase.

2.3 Differential Scanning Calorimeter and Polarization Microscopy Analysis Results of Monoolein Cubic Phase

Panel (A) of FIG. 4 is a monoolein cubic phase (0/0), a monoolein cubic phase (1/0), a monoolein cubic phase (1/0.5), and a monoolein cubic phase (1/ 0.8) shows the results of differential scanning calorimetry. In the case of the monoolein cubic phase (0/0), an endothermic peak was observed around 60.2°C. It is known that a cubic to hexagonal transition occurs when the monoolein cubic phase is heated. Therefore, the endothermic peak of the monoolein cubic phase (0/0) is determined to mean the temperature at which the cubic phase-hexagonal phase transition occurs. In the case of the monoolein cubic phase (1/0), it was confirmed that the endothermic peak was shown around 52.0°C, and the cubic phase-hexagonal phase transition was observed at 8°C lower than that of the monoolein cubic phase (0/0).

Alginate and steryl trimethyl ammonium chloride are contained in the monoolein cubic phase (1/0), but not in the monoolein cubic phase (0/0). Alginate is water soluble and is located in the water channel on a cubic monoolein. Therefore, the alginate may have only a small effect on the cubic phase transition and monoolein packing of the double layer.

On the other hand, the steryl trimethyl ammonium chloride has amphiphilic properties, so that it may be present by being inserted into the monoolein bilayer, and may flow along the bilayer, thereby lowering the phase transition temperature.

In the case of the monoolein cubic phase (1/0.5) and the monoolein cubic phase (1/0.8), it was confirmed that the phase transition occurred at 49.4°C and 48.2°C, respectively. Although there is no significant difference in the phase transition temperature, it is determined to be about 1 to 3°C lower than that of the monoolein cubic phase (1/0). Cystamine molecules electrostatically interact with alginate chains and can cross-link to form chains. Thus, most of the cystamine molecules stay in the water channel along with the alginate. Because of the low water solubility of cystamine, some cystamine molecules can get caught between the fat bilayers and lower the phase transition temperature.

Panel (B) of FIG. 4 is a differential scanning calorimeter of a monoolein cubic phase (1/0/EGF), a monoolein cubic phase (1/0.5/EGF), and a monoolein cubic phase (1/0.8/EGF). Shows the measurement result. In the case of monoolein cubic phase (1/0/EGF), monoolein cubic phase (1/0.5/EGF), and monoolein cubic phase (1/0.8/EGF), 51.6°C, 49.3°C, and 48.2°C, respectively It is confirmed that a phase transition occurs in. The phase transition temperature is a differential scanning calorimeter of a monoolein cubic phase (1/0), a monoolein cubic phase (1/0.5), and a monoolein cubic phase (1/0.8) shown in panel (A) of FIG. It is judged to be similar as a result of the measurement. Therefore, it is judged that the change in the phase transition of the monoolein cubic phase by the epithelial cell growth factor is insignificant. As described above, the change in the phase transition of the monoolein cubic phase by the epithelial cell growth factor is considered to be insignificant because the epithelial cell growth factor is water-soluble and is located only inside the water channel.

5 is a polarization optics for a monoolein cubic phase (0/0), a monoolein cubic phase (1/0), a monoolein cubic phase (1/0.5), and a monoolein cubic phase (1/0.8). Shows the results of microscopic observation.

As a result of heating the monoolein cubic phase (0/0) to 59.3°C or higher, it was confirmed that a birefringence pattern was observed. In general, the cubic phase is optically isotropic, while the hexagonal phase is optically anisotropic. The optical anisotropy appears as a birefringent pattern in a polarizing microscope. Therefore, when the monoolein cubic phase is heated above the temperature, it means that the cubic phase-hexagonal phase transition occurs. According to the observation results under a polarizing microscope, the phase transition temperatures of the monoolein cubic phase (1/0), the monoolein cubic phase (1/0.5), and the monoolein cubic phase (1/0.8) were 52.1°C, 50.2°C, respectively, And 48.8°C.

In the same way as the differential scanning calorimeter measurement result, in the observation result with a polarizing optical microscope, it was confirmed that steryl trimethyl ammonium chloride and cystamine lowered the phase transition temperature of the monoolein cubic phase. The phase transition of the monoolein cubic phase (1/0/EGF), the monoolein cubic phase (1/0.5/EGF), and the monoolein cubic phase (1/0.8/EGF) was 50.2°C, 49.2°C, and It was found to appear around 48.6°C. In the same way as the differential scanning calorimeter measurement result, it was confirmed that the change in the phase transition characteristics due to the epithelial cell growth factor was insignificant in the observation result of a polarizing optical microscope.

2.4 Measurement of Epithelial Cell Growth Factor Release

First, the release profile of the epidermal growth factor (EGF) mounted on a monoolein cubic phase (0/0/EGF) that did not contain alginate and cystamine was measured. The amount of epithelial cell growth factor released from the cubic phase after adding the monoolein cubic phase (0/0/EGF) to a release solution having a pH of 4.0, 5.5 or 7.4 and a reduced glutathione (GSH) of 0mM, 2mM or 10mM It was carried out by a method of measuring.

As a result of the experiment, most of the release was confirmed during the first 5 hours, and almost no release was observed after 5 hours. The emission pattern of the monoolein cubic phase (0/0/EGF) was confirmed to have a typical saturated emission pattern, and the emission amount was less than 2%. The reason is considered to be that the epithelial cell growth factor has a size similar to that of a monoolein cubic water channel and thus has low diffusivity.

The concentration of reduced glutathione was fixed and the pH value was changed to measure the amount of epithelial growth factor released. As a result of the experiment, it was confirmed that the amount of epithelial growth factor released was hardly affected by the change in pH.

For example, as a result of measuring the amount of release of epithelial growth factor after fixing the concentration of reduced glutathione in the release solution to 0 mM and adjusting the pH to 4.0, 5.5 and 7.4, the release amount (%) for 72 hours was 1.3% ( pH 4.0), 1.6% (pH 5.5) and 1.6% (pH 7.4).

The result is considered to be because the monoolein used in the monoolein cubic phase (0/0/EGF) has an electrically neutral characteristic. Since the monoolein has little electrical properties, it hardly performs electrostatic coupling with epithelial cell growth factors. Therefore, the amount of epithelial cell growth factor released by the change of the pH value hardly changes.

In the monoolein cubic phase (0/0/EGF), it was confirmed that the effect of the concentration of reduced glutathione on the release amount (%) of epithelial growth factor was also insignificant. The monoolein cubic phase (0/0/EGF) does not contain any reducible substances. Therefore, the amount of release (%) of the epithelial cell growth factor mounted on the cubic phase is hardly affected by a reducing factor such as reduced glutathione.

Release of epithelial cell growth factor after addition to a release solution having a reduced glutathione concentration of 0mM, 2mM, or 10mM and a pH of 4.0, 5.5, or 7.4 for the monoolein cubic phase (1/0/EGF) containing alginate The profile was analyzed.

As a result of the experiment, the amount of release (%) of epithelial growth factor in the monoolein cubic phase (1/0/EGF) was similar to the result of the monoolein cubic phase (0/0/EGF). Was confirmed.

First, as a result of confirming the change in the amount of release of the monoolein cubic phase (1/0/EGF) due to a reducing factor such as reduced glutathione, it was confirmed that there was no change in the amount of release due to reduced glutathione. This is believed to be because the monoolein cubic phase (1/0/EGF) does not contain a material that can be reduced.

On the contrary, it was confirmed that the change of the pH value affects the amount of epithelial growth factor released in the cubic phase (1/0/EGF) of monoolein. For example, in the case of a release solution with a concentration of reduced glutathione of 0 mM and a pH of 4.0, 5.5, or 7.4, the amount of epithelial growth factor released in the cubic phase (1/0/EGF) of monoolein is 1.4% (pH4.0, respectively). ), 2.1% (pH5.5), or 2.7% (pH7.4).

When the pH value of the release solution is lower than the isoelectric point of the epithelial cell growth factor, the epithelial cell growth factor has a positive charge. The positively charged epithelial cell growth factor electrically binds to the negatively charged polysaccharide alginate, which is referred to as the coacervation complex of the epithelial cell growth factor and alginate. When the epithelial growth factor and alginate are coacervated, the amount of release (%) decreases. On the other hand, when the pH value of the release solution rises above the isoelectric point of the epithelial cell growth factor, the epithelial cell growth factor has a negative charge and the binding force with alginate decreases. According to the results of panel (B) of FIG. 2, it was confirmed that the rate of coacervation complex between the epithelial cell growth factor and alginate was rapidly decreased in the pH environment above the isoelectric point of the epithelial cell growth factor. The above result is consistent with the increase in the amount of epithelial growth factor released (%) as the pH of the release solution increases.

Figure 6 is a monoolein cubic phase (1/0.8/EGF) containing alginate, cystamine, and epithelial cell growth factor is released with a reduced glutathione concentration of 0mM, 2mM, or 10mM and a pH of 4.0, 5.5, or 7.4 The results of analyzing the profile of the release of epithelial growth factors in the solution are shown.

As a result of the analysis, similar to the monoolein cubic phase (1/0/EGF), it was confirmed that the amount of epithelial growth factor released (%) also increased as the pH increased. For example, as a result of observing the release of epithelial growth factor for 72 hours in a release solution having a concentration of reduced glutathione of 0 mM and a pH of 4.0, 5.5 and 7.4 for the monoolein cubic phase (1/0/EGF), The amount of epithelial growth factor released was found to be 1.1% (pH4.0), 1.7% (pH5.5) and 2.7% (pH7.4), respectively.

As described above, when the pH is increased from 4.0 to 7.4, the electrostatic binding force between the epithelial cell growth factor and alginate decreases, and thus the release of the epithelial cell growth factor is promoted.

The monoolein cubic phase (1/0.8/EGF) differs from the monoolein cubic phase (1/0/EGF), and the release rate of the loaded epithelial cell growth factor according to the concentration of reduced glutathione contained in the release solution (% ) Was confirmed to change. For example, if the pH of the release solution is fixed to 4.0 and then the concentration of reduced glutathione is adjusted to 0, 2, and 10 mM, respectively, the release rate (%) of the epithelial growth factor released for 72 hours is 1.1% (0 mM), respectively. , 1.6% (2mM) and 2.3% (10mM).

The monoolein cubic phase (1/0.8/EGF) contains both cystamine and alginate. The cystamine contains two terminal amino groups, and the amino group at the terminal is bonded to alginate through electrostatic bonding to form an alginate-cystamine complex (see FIG. 2(A)). The alginate-cystamine complex is in a gel state and constrains the epithelial cell growth factor.

However, when the monoolein cubic phase (1/0.8/EGF) is present in the release solution containing reduced glutathione, the reducing factor in the solution penetrates into the cubic phase water channel, resulting in a disulfide bond of cystamine. Is removed, which expands the space inside the water channel and promotes the diffusion of epithelial cell growth factors, thus increasing the amount of release (%).

In summary, since alginate is contained in the monoolein cubic phase, the release of the epithelial cell growth factor mounted on the cubic phase is suppressed in an acidic environment, and when the pH increases, the release of the epithelial cell growth factor mounted on the cubic phase is suppressed. Is increased.

In addition, in the monoolein cubic phase containing both alginate and cystamine, the disulfide bonds of cystamine are removed by reduced glutathione, thereby expanding the space of the water channel, so that the amount of release of the epithelial cell growth factor mounted in the cubic phase increases.

The drug delivery system must safely store the drug until it is injected into the body, and the release of the drug must be enhanced after it is injected into the body. It is determined that the cubic phase monoolein of the present invention meets the performance of the drug delivery system.

The monoolein cubic phase of the present invention is characterized in that the release of epithelial cell growth factors is suppressed in a non-reducing state or a pH environment below the isoelectric point.

In contrast, the monoolein cubic phase containing both alginate and cystamine of the present invention promotes the release of the loaded epithelial cell growth factor in a pH environment above the isoelectric point or in a reduced state, but all of the above environments are similar to those in the human body. There is an advantage that it can be used as a drug delivery system.

3. Conclusion

The present invention relates to a monoolein cubic phase containing alginate and cystamine in a water channel. The cubic phase of the present invention is characterized in that the amount of release of the epithelial cell growth factor mounted therein is controlled according to the pH value and reduced state of the release solution.

According to the light scattering intensity measurement results of the alginate-cystamine mixed solution, it was confirmed that the maximum degree of crosslinking between alginate and cystamine at pH 3.0 was when the molar ratio of carboxyl group and amino group was 1:1.5. In addition, it was confirmed that the formation of the coacervate complex between alginate and epithelial cell growth factor was best made at pH 4.2, where the electrostatic interaction between alginate and epithelial cell growth factor was the best.

As a result of observation through an electron microscope, it was confirmed that the monoolein cubic phase was successfully prepared. As a result of differential scanning calorimetry (DSC) and polarization microscopy observations, the phase transition temperature of the monoolein cubic phase is characterized by the addition of steryl trimethyl ammonium chloride, alginate, and cystamine, but the epithelial cell growth factor It was confirmed that it did not change.

It was confirmed that the release amount (%) of the epithelial cell growth factor mounted on the cubic phase reached the maximum after 5 hours, and the release amount (%) of the epithelial cell growth factor was found to be 1.4 to 4.1%.

Since the monoolein cubic phase of the present invention contains alginate and cystamine in the water channel, when the pH is increased and the reduced glutathione concentration is increased, the binding of the epithelial cell growth factor and alginate is reduced, and the disulfide bond of cystamine is reduced. Since it is reduced and disappears, the release rate (%) of the loaded epithelial cell growth factor is increased. In addition, in the release environment where the isoelectric point of the epithelial cell growth factor is less than pH 4.6 or in a non-reducing state, the mounted epithelial cell growth factor is not released and is safely stored inside the water channel.

Therefore, when the cubic phase monoolein of the present invention is in a pH state above the isoelectric point of the epithelial cell growth factor, and is injected into the body where the reduced state is always maintained, the epithelial cell growth factor mounted therein is released. This means that the cubic phase of nomoolein containing alginate and cystamine of the present invention can be used as a drug delivery system for carrying a peptide drug or a protein drug.

The specific embodiments described herein are meant to represent preferred embodiments or examples of the present invention, and the scope of the present invention is not limited thereby. It will be apparent to those skilled in the art that variations and other uses of the invention do not depart from the scope of the invention described in the claims of this specification.

Claims (10)

A monooleion nanostructure for cosmetic materials for delivery of active ingredients to the skin and body, including a polysaccharide-disulfide compound complex.
The method of claim 1, wherein the monoolein nanostructure is a monoolein cubic phase (monoolein cubic phase), characterized in that the monoolein (monooleion) nanostructure for a cosmetic material for delivery of active ingredients to the skin and body.
The method of claim 2, wherein the monoolein cubic phase has steryl trimethyl ammonium chloride inserted in a monoolein bilayer. Monooleion nanostructure.
The method of claim 1, wherein the polysaccharide-disulfide compound complex is fixed to the double layer constituting the water channel of the monooleion nanostructure for drug delivery in the body. Monooleion nanostructure.
The method of claim 1, wherein the polysaccharide-disulfide compound complex is composed of a polysaccharide containing one or more carboxyl groups and a disulfide compound having one or more amino groups.Monooleion for cosmetic materials for delivery of active ingredients to skin and body. ) Nanostructure.
[2] The nanostructure of claim 1, wherein the polysaccharide-disulfide compound complex forms a three-dimensional network by disulfide bonds.
The method of claim 1, wherein the monooleion nanostructure for a cosmetic material for delivery of active ingredients to skin and body has an isoelectric point of pH 4 to 6 in the polysaccharide-disulfide compound complex as the active ingredient. A monooleion nanostructure for cosmetic material for delivery of active ingredients to skin and body, characterized in that it further comprises a physiologically active substance.
The method of claim 7, wherein the physiologically active substance is present in a solution in which a monooleion nanostructure for a cosmetic material for delivery of active ingredients to the skin and body is at pH 7 to 8 and a concentration of reduced glutathione is 1 to 10 mM In the case of, a monooleion nanostructure for a cosmetic material for delivery of active ingredients to skin and body, characterized in that diffusion rate is improved by diffusion from the inside of the polysaccharide-disulfide compound complex to the water channel.
A first step of mixing monoolein and steryl trimethyl ammonium chloride at 45 to 55°C and cooling to 30 to 40°C to prepare a mixed melt;
The second step of preparing a monooleion nanostructure by adding a polysccharide, a disulfide compound, and a physiologically active substance while maintaining the mixed melt at 30 to 40°C and maintaining it at 20 to 25°C. ;
A method of manufacturing a monooleion nanostructure for a cosmetic material for delivery of active ingredients to the skin and body comprising a.
The method of claim 9, wherein the monoolein and steryl trimethyl ammonium chloride are mixed in a molar ratio of 1:4 to 6, and the polysccharide and the disulfide compound are the carboxyl group of the polysaccharide and the amino group of the disulfide compound. A method of manufacturing a monooleion nanostructure for cosmetic materials for delivery of active ingredients to the skin and body, characterized in that the mixture is mixed in a molar ratio of 1:0.6 to 1:1.

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