KR20120092804A - Anionic natural polymer and amphoteric natural polymer-immobilized cubic phase and method for preparing the same - Google Patents

Abstract

PURPOSE: A monoolein cubic phase on which alginate and silk fibroin are fixed is provided to release less target ingredients at low pH and release target ingredient a lot of target ingredients. CONSTITUTION: A monoolein cubic phase contains anionic natural polymers containing alginate and amphoteric natural polymers containing silk fibroin which are fixed on an aqueous channel thereinside. The alginate and silk fibroin are conjugated by a hydrophobic anchor. The hydrophobic anchor includes monoalakyl acrylate, dialkyl acrylate, palmitic acid, stearyl amine, or stearic acid. A method for preparing the monoolein cubic phase comprises: a step of mixing the silk fibroin and target materials to prepare a mixture solution; and a step of adding the mixture solution to a monoolein melt solution to form transparent gel.

Description

Cubic phase to which anionic natural polymer and amphoteric natural polymer are immobilized and a method for preparing the same {ANIONIC NATURAL POLYMER AND AMPHOTERIC NATURAL POLYMER-IMMOBILIZED CUBIC PHASE AND METHOD FOR PREPARING THE SAME}

The present invention relates to a monooleic cubic phase in which an anionic natural polymer and an amphoteric natural polymer are immobilized and a method for preparing the same, and specifically, a hydrophobized anionic natural polymer and a hydrophobic amphoteric natural polymer are immobilized therein. It relates to a cubic phase and a method for producing the same.

Glycerol monoolate, which is a packing parameter close to 1, or monooolein, forms a cubic phase if the temperature of the system in the water phase, the concentration of monoolein and the content of the water phase are appropriate. The cubic phase is transparent because it has an organized isotropic structure.

The monoolein constituting the cubic phase is not toxic to the human body and is suitable for use in a living body and is biodegradable. It also exhibits sustained-release behavior that slowly releases the drug and has excellent adhesion to the skin and mucous membranes, resulting in injection, transdermal, trans-mucous membrane delivery and other drug delivery within the human body. Actively studied as carriers [JC Shah, Y Sadhale, DMChilukuri, Cubic phase gels as drug delivery systems, Adv. Drug Deliv. Rev. 47 (2001) 229-250.

On the other hand, as an example of the patent technology related to the monoolein cubic phase capable of trapping the target material, there is a Korean Patent Publication No. 10-1991-0009242 (published: June 28, 1991). The patent claims that 'oral disease, consisting essentially of 1 to 99% of monoolein and an anti-inflammatory agent, antimicrobial agent, antibiotic, peroxide, anesthetic and vitamin, and 1 to 90% of active agent suitable for the treatment of oral disease. Disclosed are compositions suitable for infusion into or around the periodontal capsule of a suffering person or lower animal.

However, the monoolein cubic phase disclosed in the above publication has the effect of slowly releasing the trapped drug by simple diffusion, and does not mention the technical idea of selectively releasing the target component collected therein under specific conditions.

It is an object of the present invention to provide a monooleic cubic phase immobilized with a natural polymer capable of releasing the desired substance at certain pH conditions.

Another object of the present invention is to provide a method for producing a monoolefin cubic phase to which the natural polymer is immobilized.

The monoolein cubic phase of the present invention for achieving the above object is an anionic natural polymer and an amphoteric natural polymer is immobilized in an inner water channel.

Alginate may be used as the anionic natural polymer and silk fibroin may be used as the amphoteric natural polymer.

In one embodiment, the alginate and the silk fibroin may be immobilized by binding a hydrophobic anchor capable of providing a hydrophobic bond as an immobilizing material between the aqueous channel and the alginate and the silk fibroin.

In one embodiment, the hydrophobic anchor in the group consisting of monoalkylacrylate, dialkylacrylate, dialkylacrylate, palmitic acid, stearylamine and stearic acid It includes at least one selected and may have a carbon number of C ₁₂ ~ C ₂₂ ranges.

In one embodiment, the weight ratio of the alginate and the hydrophobic anchor may range from 1000: 1 to 20: 1, and the weight ratio of the silk fibroin and the hydrophobic anchor may range from 500: 1 to 10: 1.

In one embodiment, the weight ratio of the immobilized alginate and the immobilized silk fibroin may range from 1: 5 to 1: 100.

Another object of the present invention described above is to prepare a mixed solution by mixing an alginate / immobilized material combined silk fibroin and a target material to be immobilized, and adding the mixed solution to a monoolefin melt to produce a transparent gel. It is achieved by a process for the preparation of a monooleic cubic phase comprising the step of forming.

In one embodiment, the temperature of the monoolein melt may be in the range of 30 ~ 60 ℃.

In one embodiment, the monoolein melt may be obtained by melting in a bath.

In one embodiment, the mixing ratio of the mixed solution and the monoolein melt may range from 3: 2 to 1: 3 by weight.

In one embodiment, the weight ratio of the immobilized alginate and the immobilized silk fibroin may range from 1: 5 to 1: 100.

In one embodiment, the concentration of the alginate to which the immobilization material is bound and the silk fibroin to which the immobilization material is bound may range from 1 to 15% in the mixed solution.

In one embodiment, the alginate to which the immobilization material is bound is prepared by dissolving the immobilization material in an organic solvent to prepare an immobilization material solution, dissolving alginate in water to prepare an alginate aqueous solution, and mixing the two solutions. It may be prepared through the step of.

In one embodiment, the silk fibroin combined with the immobilization material, dissolving the silk fibroin in water to prepare a silk fibroin aqueous solution, dissolving the immobilized material in the silk fibroin aqueous solution to prepare a mixed solution and the mixing The solution may be prepared by a step of basicizing and reacting.

In the present invention, an anionic natural polymer comprising alginate and an amphoteric natural polymer comprising silk fibroin are anticancer agents, antibiotics, antifungal agents, antibacterial agents and antioxidants as a target substance in a monoolein cubic phase immobilized on an inner water channel. It provides a system comprising at least one of a whitening agent and an anti-inflammatory agent.

The monoolein cubic phase according to the present invention has a structure in which hydrophobized alginate and hydrophobized silk fibroin are immobilized on the microaqueous channel of the cubic phase. Because it prevents the release of a relatively small amount of the collected target component, and the complex coacervate dissolves at a pH higher than the isoelectric point of silk fibroin, it exhibits a relatively large release of the captured target component.

1A is a schematic diagram of a cubic phase in which alginate to which immobilization material is bound and silk fibroin with immobilization material are immobilized, and a complex coacervate is formed when the pH of the system is lower than the isoelectric point of silk fibroin.
FIG. 1B is a schematic diagram of a cubic phase in which alginate to which immobilization material is bound and silk fibroin with immobilization material are immobilized, and a complex coacervate is dissolved when the pH of the system is higher than the isoelectric point of silk fibroin.
2 shows the FT-IR spectrum for alginate (b) in which alginate (a) and immobilization material are bound.
Figure 3 shows the FT-IR spectrum for silk fibroin (b) combined with silk fibroin (a) and the immobilization material.
FIG. 4 is a graph showing the average diameter (rod) and light scattering intensity (•) of the silk fibroin complexes to which the alginate / immobilization materials to which immobilization materials formed at pH 3.0 are bound.
FIG. 5 is a graph showing the results of observing the average diameter (rod) and light scattering intensity (•) of the silk fibroin composite in which the alginate / immobilization material is bonded to the immobilization material.
FIG. 6 is a graph showing the results of observing FITC-dextran release from a cubic phase containing no silk fibroin bound to immobilized material / alginate / immobilized material bound with varying pH.
FIG. 7 is a graph showing the results of observing FITC-dextran release from a cubic phase containing silk fibroin bound to immobilized material / alginate / immobilized material bound with varying pH.

Hereinafter, with reference to the accompanying drawings will be described in detail with respect to the monoolein cubic phase and its preparation method according to an embodiment of the present invention. While the invention is susceptible to various modifications and alternative forms, specific embodiments thereof are shown by way of example in the drawings and will herein be described in detail. It is to be understood, however, that the invention is not intended to be limited to the particular forms disclosed, but on the contrary, is intended to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the invention. In the accompanying drawings, the dimensions of the structures are enlarged to illustrate the present invention in order to clarify the present invention.

The terminology used herein is for the purpose of describing particular example embodiments only and is not intended to be limiting of the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In this application, the terms "comprise" or "having" are intended to indicate that there is a feature, number, step, operation, component, or a combination thereof described in the specification, but one or more other features or numbers. It is to be understood that the present invention does not exclude in advance the possibility of the presence or the addition of steps, actions, components, or a combination thereof.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms such as those defined in the commonly used dictionaries should be construed as having meanings consistent with the meanings in the context of the related art and shall not be construed in ideal or excessively formal meanings unless expressly defined in this application. Do not.

Hereinafter, the present invention will be described in detail.

Alginate is an anionic natural polymer and silk fibroin is a protein that exhibits positive charges at acidic levels lower than the isoelectric point (PI) and negative charges at neutral and basic levels higher than the isoelectric point. Thus, at acids below the isoelectric point, alginate and silk fibroin form complex coacervates due to electrostatic attraction.

On the other hand, when the pH increases higher than the isoelectric point and is neutral or basic, the complex coacervate dissolves because the silk fibroin shows a negative charge. Immobilization of alginate and silk fibroin, which exhibit such complex coacervate properties, to the aqueous channel of the cubic phase results in different properties depending on the pH change.

If the pH of the system is lower than the isoelectric point of silk fibroin, the complex coacervate is formed, which closes the aqueous channel, thereby releasing a relatively small amount of the desired component. On the other hand, when the pH of the system is higher than the isoelectric point of silk fibroin, the complex coacervate dissolves, leading to the opening of the aqueous channel, thereby rapidly increasing the rate of release of the desired component.

1A is a schematic diagram of a cubic phase in which alginate to which immobilization material is bound and silk fibroin with immobilization material are immobilized, and a complex coacervate is formed when the pH of the system is lower than the isoelectric point of silk fibroin.

Referring to FIG. 1A, when the pH is lower than the isoelectric point of silk fibroin, alginate with immobilized material and silk fibroin with immobilized material form a complex coacervate, which closes the aqueous channel and releases the target material, FITC-dextran. It can be confirmed that this is suppressed.

FIG. 1B is a schematic diagram of a cubic phase in which alginate to which immobilization material is bound and silk fibroin with immobilization material are immobilized, and shows a case where complex coacervate is dissolved when the pH of the system is higher than the isoelectric point of silk fibroin.

Referring to FIG. 1B, when the pH of the system increases higher than the isoelectric point of silk fibroin and is neutral or basic, complex coacervate dissolves because silk fibroin exhibits a negative charge. As a result, the release amount of the target substance FITC-dextran is increased.

1A and 1B, according to the monoolein cubic phase according to the present embodiment, it can be seen that the emission amount of the target component collected according to the pH value varies.

On the other hand, in the monooleic cubic phase, alginate and silk fibroin can be immobilized to the water channel inside the cubic phase through various methods known as immobilization means. For example, a hydrophobic anchor may be bonded between the aqueous channel and alginate and silk fibroin to provide a hydrophobic bond as an immobilization material.

Specific examples of hydrophobic anchors include at least one of monoalkylacrylate, dialkylacrylate, palmitic acid, stearylamine, and stearic acid. . It is also appropriate that the hydrophobic anchor has a C ₁₂ to C ₂₂ range. A hydrophobic anchor is a substance capable of providing a hydrophobic bond, which is a kind of immobilization means, between the water channel inside the cubic phase and alginate or silk fibroin. It will be fixed through.

Specifically, the weight ratio of the silk fibroin and the hydrophobic anchor may be in the range of 500: 1 to 10: 1, and the weight ratio of the immobilized alginate and the immobilized silk fibroin is in the range of 1: 5 to 1: 100. It is also easily applicable.

Hereinafter, a method of manufacturing a cubic phase according to an embodiment of the present invention will be described in detail.

According to the manufacturing method of the monoolein cubic phase, a mixed solution is prepared by mixing a silk fibroin in which an immobilization material is bonded to be immobilized in an aqueous channel inside the cubic phase, and a target material to be collected in the water channel; Adding the mixture to a monoolein melt to form a transparent gel.

In the manufacturing method of the cubic phase according to the embodiment of the present invention, the temperature of the monoolefin melt is preferably in the range of 30 ~ 60 ℃. If the temperature is lower than 30 ℃ monoolein may not melt, because if the temperature is higher than 60 ℃ monoolein may be oxidized and denatured. The temperature of the monoolein melt is more preferably 35 to 55 ℃, most preferably 40 to 50 ℃.

In addition, since monoolein is a monoglyceride containing unsaturated hydrocarbons, it can be chemically oxidized at high temperatures, thereby breaking the hydrocarbon chain. Therefore, when melting monoolein, it is advisable to gradually increase the temperature or to melt it in a bath to prevent a sudden rise in temperature. More preferably, the temperature of the mixed solution is the same as the temperature of the monoolein melt.

In the method for producing a monoolein cubic phase, the ratio of the mixed solution and the monoolein melt is preferably in the range of 3: 2 to 1: 3 as weight ratio. If the amount is lower than this range, the amount of the mixed solution may not be sufficient to hydrate the monoolein completely. Therefore, the cubic phase may not be formed. If the amount is higher than this range, the mixed solution may be excessively large due to the excessive amount of the mixed solution. It may not be all absorbed. The ratio of the mixed solution and the monoolein melt is more preferably 1: 1 to 1: 2 by weight ratio, and most preferably 4: 5 to 3: 5.

In the method for producing a monoolein cubic phase, the weight ratio of alginate to which immobilization material is bound and silk fibroin to which immobilization material is bound is 1: 5 to 1: 100. Outside of this range, the amount of complex coacervate produced may be too small because of too much negative charge of alginate or too much positive charge of silk fibroin. Preferably 1: 10-1: 50, Most preferably, 1: 15-1: 30.

The total concentration of alginate with immobilized material and silk fibroin with immobilized material is preferably in the range of 1-15% in the mixed solution. At higher concentrations than 15%, the viscosity of the solution may be too high to produce cubic phases, and at concentrations lower than 1%, the cubic phases contain very small amounts of alginate and silk fibroin, resulting in inadequate pH-sensitive release properties. Because. More preferably, the total concentration of the alginate to which the immobilization material is bound and the silk fibroin to which the immobilization material is bound is 1 to 10%, most preferably 1 to 3% in the mixed solution.

In the monoolein cubic phase manufacturing method of the present invention, the alginate to which the immobilization material is bound is preferably prepared by dissolving the immobilized material in an organic solvent to prepare a solution of the immobilized material, dissolving alginate in water to prepare an alginate aqueous solution, and The two solutions may be prepared by mixing and reacting the two solutions. The reaction mixture obtained may be further filtered to collect the reactants.

The immobilization material is preferably a hydrophobic anchor, more preferably the hydrophobic anchor is preferably added so that the alginate and the hydrophobic anchor are in a weight ratio of 1000: 1 to 20: 1. If the content of the hydrophobic anchor is lower than 1000: 1, the hydrophobization of alginate may be insufficient, and thus the alginate may not be properly immobilized in the cubic phase aqueous channel. If the content of the hydrophobic anchor is higher than 20: 1, it is not preferable because the water solubility of the alginate is reduced. More preferably, the hydrophobic anchor is an alginate and a hydrophobic anchor is added in a weight ratio of 500: 1 to 30: 1, most preferably 200: 1 to 50: 1.

Hydrophobic anchors serve to immobilize alginates because they are inserted into the hydrocarbon matrix on the cubic phase. As the hydrophobic anchor, a fatty acid is preferable, and the carbon number of the fatty acid is preferably C ₁₂ to C ₂₂ . If the carbon number is less or more than this range, it is difficult to act as a hydrophobic anchor. The carbon number of the fatty acid is more preferably C ₁₄ ~ C ₂₀ , most preferably C ₁₆ ~ C ₁₈ .

In the monoolein cubic phase manufacturing method of the present invention, the silk fibroin to which the immobilization material is bonded is prepared by dissolving silk fibroin in water to prepare a silk fibroin aqueous solution, dissolving the immobilized material in a silk fibroin aqueous solution mixed solution It may be prepared through the step of preparing, reacting the mixture by basifying. The step of removing unreacted material from the reaction mixture obtained may be further performed.

The immobilization material is preferably a hydrophobic anchor, more preferably the hydrophobic anchor is preferably added so that the silk fibroin and the hydrophobic anchor are in a weight ratio of 500: 1 to 10: 1. The low content of hydrophobic anchors results in a lack of hydrophobization of silk fibroin, which may prevent silk fibroin from properly immobilizing the cubic phase aqueous channel. High content of hydrophobic anchors can reduce the water solubility of silk fibroin. Even more preferably, the hydrophobic anchor is preferably added with silk fibroin and a hydrophobic anchor in a weight ratio of 200: 1 to 10: 1, most preferably 50: 1 to 10: 1.

Hydrophobic anchors serve to immobilize silk fibroin because they are inserted into the hydrocarbon matrix on the cubic phase. Hydrophobic anchors are preferably fatty acids and derivatives thereof, and the carbon number of the hydrocarbon chains of the fatty acids and derivatives thereof is preferably C ₁₂ to C ₂₂ . This is because if the carbon number is less or more than this range, it does not act as a hydrophobic anchor. At this time, the carbon number of the hydrocarbon chain of the fatty acid and fatty acid derivative is more preferably C ₁₄ ~ C ₂₀ , most preferably C ₁₆ ~ C ₁₈ .

As the target substance to be collected in the cubic phase, various hydrophobic and hydrophilic bioactive components such as anticancer agents, antibiotics, antifungal agents, antibacterial agents, antioxidants, whitening agents and anti-inflammatory agents may be used.

Hereinafter, the present invention will be described in more detail with reference to specific examples.

Immobilized material Combined Alginate  Produce

Stearyl acid was dissolved in 10 ml of propanol so that the concentration of the solution was 0.026% (w / v). The alginate was then dissolved in 10 ml of distilled water to a concentration of 1.0% (w / v). The reaction was prepared by mixing the stearyl acid solution and alginate solution prepared above in a 50 ml beaker and stirred at 50 ℃ for 12 hours. After the reaction was completed, the reaction mixture was filtered using filter paper (Whatman No. 2), and the cake (cake, alginate with immobilized material) was dried at 40 ° C. for 48 hours in a vacuum oven.

Immobilized material Combined Alginate FT - IR  Spectral observation

KBr pellets containing alginate with immobilized material were prepared and observed for FT-IR spectra (Perkin-Elmer Fourier Transformed Infrared Spectrophotometer instrument, EXCALIBER Series, U.S.A.).

2 shows the FT-IR spectrum for alginate (b) in which alginate (a) and immobilization material are bound.

Referring to Figure 2, the peak observed at 1040 cm ^-1 in the alginate spectrum (a) is due to -COC- stretching, the peak observed at 1595 cm ^-1 is due to -COOH stretching, 3233 cm ^-1 The broad peak observed at is the OH stretching signal. In the alginate (b) spectrum with immobilized material bound, the peak observed at 1245 cm ⁻¹ is due to amide III (CN) bending and the peak observed at 1380 cm ⁻¹ is the signal for -CH ₂ -of the alkyl chain. The strong peak observed at 1580 cm ⁻¹ is due to amide II (NH) bending and the peak observed at 1650 cm ⁻¹ is due to amide I (C═O) stretching. Since the signal of the methylene group of the alkyl chain and the signal of the amide bond were observed, it can be seen that stearylamine was successfully bound to the alginate.

Immobilized material Combined silk Fibroin  Produce

1 g of silk fibroin was dissolved in 90 ml of distilled water to prepare an aqueous silk fibroin solution. 0.01 g of 2-dodecane-1-yl succinic anhydride {(2-dodecen-1-yl) succinic anhydride} was dissolved in an aqueous silk fibroin solution. After adjusting the pH of the reaction solution to 10.0 using 1N NaOH aqueous solution, the reaction was performed while maintaining the pH at 10.0 for 5 hours at 70 ° C. After the reaction mixture was cooled to room temperature, 1N HCl was added to adjust the pH to 7.0. Unreacted anhydride was extracted and removed using methyl tert-butyl ether. The reaction solution from which the unreactant was removed was freeze-dried to obtain silk fibroin to which immobilized material was bound.

Immobilized material Combined silk Fibroin FT - IR  Spectral observation

KBr pellets containing silk fibroin with immobilized material were prepared to observe the FT-IR spectra.

Figure 3 shows the FT-IR spectrum for silk fibroin (b) combined with silk fibroin (a) and the immobilization material.

Referring to Figure 3, the silk fibroin spectrum (a) at, 1250 cm ^-1, 1531cm ^-1, a strong peak at 1644cm ^-1 are observed each amide III (CN) stretching, amide II (NH) bending, amide I ( C = O) stretch. Characteristic peaks of amide bonds were also observed in silk fibroin (b) to which immobilization material was bound, and CH ₃ bending of alkyl chains of 2-dodecane-1-yl succinic acid residues was observed at 1375 cm ⁻¹ . It means that can-1-yl succinic acid group was successfully bound to silk fibroin.

Immobilized material Combined With alginate  Immobilized material Combined silk Fibroin  complex Coacervate  observe

Alginate with immobilized material and silk fibroin with immobilized material were dissolved in glycine buffer (pH 3.0). In this case, the concentration of the total polymer was 1%, and the mixing ratio of the alginate / immobilization material-bonded silk fibroin combined with the immobilization material was varied in a range of 1: 1 to 1:35 by weight ratio. The size and light scattering intensity of the complex coacervate, a complex formed from the mixture, was determined by particle analyzer (ZetaPlus 90, Brookhaven Instrument Co., USA). To examine the effect of pH on complex coacervation, alginate with immobilized material and silk fibroin with immobilized material were mixed with several glycine buffers (pH 3.0, pH 4.0, pH 4.5). , MES buffer (pH 5.0), HEPES buffer (pH 7.0), Trizma base buffer (pH 9.0)).

FIG. 4 is a graph showing the average diameter (rod) and light scattering intensity (•) of the silk fibroin complexes to which the alginate / immobilization materials to which immobilization materials formed at pH 3.0 are bound.

Referring to FIG. 4, when the ratio of the alginate to which the immobilization material is bound and the silk fibroin to which the immobilization material is bound is 1:25, the average diameter was 580 nm, and in all ratios except this case, the average diameter was 300 to 400 nm. Light scattering intensity also reached its maximum at a 1:25 ratio.

The diameter of the composite is a measure of the number of polymers that make up the composite particles, and the light scattering intensity is proportional to the number of composite particles as well as the size. Therefore, it can be seen that the maximum complex coacervation occurs when the ratio of the alginate to which the immobilization material is bound and the silk fibroin to which the immobilization material is bound is 1:25.

FIG. 5 is a graph showing the results of observing the average diameter (rod) and light scattering intensity (•) of the silk fibroin composite in which the alginate / immobilization material is bonded to the immobilization material. FIG. 5 shows the results for the composite obtained when the ratio of alginate to which immobilization material is bound and silk fibroin with immobilization material is 1:25.

Referring to FIG. 5, the average diameter and the light scattering intensity decreased sharply as the pH increased from 3 to 5. As the pH increases, the silk fibroin with immobilized material becomes negatively charged, so that the intermolecular electrostatic attraction disappears and the complex dissolves. As such, the complex has a tendency to dissolve as the pH is increased, so that the size and light scattering intensity are reduced.

Cubic tops  Produce

2 g of monoolein (MO) was placed in a vial and melted in a bath in a constant temperature water bath at 60 ° C. The alginate combined with the immobilized material and the silk fibroin combined with the immobilized material were dissolved together in distilled water to prepare a mixed solution having a concentration of 0.077% and 1.924%, respectively. Then, the fluorescent dye FITC-dextran (fluorescein-isothiocyanate-conjugated dextran) was dissolved in the mixed solution so that the concentration was 0.2%. 0.858 g of the mixed solution was heated to 60 ° C. and then slowly poured over the melt of MO. The mixture solution was allowed to stand at room temperature until it was completely absorbed to form a transparent gel.

FITC - Dextran  Observation of release

5 ml of buffer solution was slowly poured onto the cubic in a 20 ml glass vial. Glycine buffer was used to observe the release at pH 3.0, pH 4.0 and pH 4.5, MES (2- (N-morpholino) -ethanesulfonim acid) buffer was used to observe the release at pH 5.0, pH 7.0 HEPES (N- [2-hydroxyethyl] piperazine-N '-[2-ethanesulfonic acid]) buffer was observed for the release at, and Trizma base buffer was used for the release at pH 9.0. In order to measure the amount of fluorescent dyes released over a period of time, 0.1 ml of the supernatant was collected. 0.1 ml of buffer solution was added to each sample to compensate for the decrease in the discharge caused by the collection of the supernatant. The amount of FITC-dextran emitted from the cubic phase was determined by exciting the fluorescent intensity of the collected supernatant at 520 nm and measuring at 492 nm.

FIG. 6 is a graph showing the results of observing FITC-dextran release from a cubic phase containing no silk fibroin bound to immobilized material / alginate / immobilized material bound with varying pH.

Referring to Figure 6, the release gradually increased over time for 100 hours. Regardless of the pH of the release solution, the% release over time was about the same.

FIG. 7 is a graph showing the results of observing FITC-dextran release from a cubic phase containing silk fibroin bound to immobilized material / alginate / immobilized material bound with varying pH.

Referring to FIG. 7, the degree of release increased with increasing pH of the discharge solution. For example, the% release for 100 hours at pH 3.0, pH 4.0, pH 4.5, pH 5.0, pH 7.0, pH 9.0 was 2.4%, 3.1%, 3.6%, 4.7%, 6.6%, 7.2%, respectively. In acidity, the degree of release is low because complex coacervates of alginate with immobilized material and silk fibroin with immobilized material are formed to block microchannels on the cubic phase. However, as the pH increases, the degree of release increases because the complex is dissolved.

The monoolein cubic phase according to the present invention has a structure in which the hydrophobized alginate and the hydrophobized silk fibroin are immobilized on the microaqueous channel of the cubic phase. Because it blocks the aqueous channel, it emits a relatively small amount of the captured target component, and the complex coacervate dissolves at a pH higher than the isoelectric point of the silk fibroin, thereby releasing a relatively large amount of the captured target component. Therefore, it is possible to control the release amount of the target substance according to the pH can be applied to a variety of uses, such as drug delivery vehicle.

As described above with reference to a preferred embodiment of the present invention, those skilled in the art or those skilled in the art without departing from the spirit and scope of the present invention described in the following specific claims It will be appreciated that various modifications and variations can be made in the present invention.

Claims (14)

A monooleic cubic phase in which an anionic natural polymer comprising alginate and an amphoteric natural polymer comprising silk fibroin are immobilized on an inner water channel. The method of claim 1, wherein the alginate and the silk fibroin is characterized in that the hydrophobic anchor (hydrophobic anchor) that can provide a hydrophobic bond as an immobilizing material between the water channel and the alginate and the silk fibroin is bonded and immobilized Mono-Olein Cubic Award. The hydrophobic anchor according to claim 2, wherein the hydrophobic anchor is monoalkylacrylate, dialkylacrylate, palmitic acid, stearylamine and stearic acid. Monooleic cubic phase comprising at least one selected and having from ₁₂ to ₂₂ carbon atoms. The weight ratio of the alginate and the hydrophobic anchor is in the range of 1000: 1 to 20: 1, and the weight ratio of the silk fibroin and the hydrophobic anchor is in the range of 500: 1 to 10: 1. Mono-Olein Cubic Prize. The monoolefin cubic phase according to claim 1, wherein the weight ratio of the immobilized alginate and the immobilized silk fibroin is in the range of 1: 5 to 1: 100. Preparing a mixed solution by mixing the alginate / immobilized material combined with silk fibroin and the target material to be collected; And
Adding the mixed solution to the monoolein melt to form a transparent gel. The method of claim 6, wherein the temperature of the monoolein melt is in the range of 30 to 60 ℃. The method according to claim 6 or 7, wherein the monoolein melt is obtained by melting in a hot bath. The method of claim 6, wherein the mixing ratio of the mixed solution and the monoolein melt is in the range of 3: 2 to 1: 3 by weight. The method of claim 6, wherein the weight ratio of the immobilized alginate and the immobilized silk fibroin is in the range of 1: 5 to 1: 100. The method of claim 6, wherein the concentration of alginate to which the immobilization material is bound and silk fibroin to which the immobilization material is bound is in the range of 1 to 15% in the mixed solution. The alginate to which the immobilization material is bound comprises:
Dissolving the immobilized material in an organic solvent to prepare an immobilized material solution;
Dissolving alginate in water to prepare an alginate aqueous solution; And
Method for producing a mixture characterized in that through the step of reacting the two solutions. The silk fibroin of claim 6, wherein the immobilization material is bound to:
Dissolving silk fibroin in water to prepare an aqueous silk fibroin solution;
Preparing a mixed solution by dissolving an immobilization material in the aqueous silk fibroin solution; And
The preparation method characterized in that it is prepared through the step of reacting the basic mixture solution. Anionic natural polymers containing alginate and amphoteric natural polymers containing silk fibroin are anticancer agents, antibiotics, antifungal agents, antibacterial agents, antioxidants, whitening agents and anti-inflammatory agents in the monooleic cubic phase immobilized on the inner water channel. A system comprising at least one of the.

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