KR101886037B1 - Temperature-responsive cubic phase containing Ion complex which exhibits temperature-dependent self-assembling property - Google Patents

Abstract

The present invention relates to a temperature responsive cubic phase prepared by immobilizing an ionic complex having temperature dependent self-aggregating properties on a cubic phase water channel. More specifically, an ionic complex formed of polyethylenimine and cinnamic acid is fixed on a monoolein cubic phase to control and release the substance placed on the cubic phase in response to temperature. The temperature-responsive cubic phase of the present invention is formed by immobilizing a polyethyleneimine / cinnamic acid ion complex, which is characterized by self-assembling and self-disassembling depending on the temperature change, And emits a release effect in response to the change.

Description

[0001] The present invention relates to a temperature-responsive cubic phase containing an ion complex having temperature-dependent self-aggregation characteristics,

The present invention relates to a temperature responsive cubic phase prepared by immobilizing an ionic complex having temperature dependent self-aggregating properties on a cubic phase water channel. More specifically, an ionic complex formed of polyethylenimine and cinnamic acid is fixed on a monoolein cubic phase to control and release the substance placed on the cubic phase in response to temperature.

Monoolein (MO) forms a bicontinuous cubic phase (hereinafter cubic phase) with amphiphiles whose packing parameters are slightly greater than 1 (CV G. Iglesias-Salto, S. Engelskirchen, S. Ahualli, Phys. Chem. Chem. Phys 13 (2011) 3004). The cubic phase has two mutually intersecting water channels (about 2-3 nm in diameter) surrounded by a mono-olein bilayer (MO bilayer, about 3.5 nm thick) matrix. The cubic phase is optically transparent because it is isotropy, and the content of water on the cubic is 28 to 40%, and is classified into a gyroid cubic phase and a diamond cubic phase depending on the content of water. The cubic phase has a merit that water-soluble compounds and lipid-soluble compounds can be loaded at the same time since the cubic phase includes a water channel and a lipid matrix, and the water-soluble compound can be mounted on the water channel and the lipophilic compound can be mounted on the matrix.

On the other hand, polyethyleneimine is a cationic polymer having an amino group, and cinnamic acid is an anionic compound having a carboxyl group. Using the Henderson-Hasselbalch'equation and the pKa value, the ionization degree of the amino group and the ionization degree of the carboxyl group are almost 100% in the aqueous solution of pH 7.0. As a result, Shinnam acid can be easily bonded to polyethylene imine through electrostatic attraction.

In addition, due to Shin-Na acid having a hydrophobic phenyl group, the polyethyleneimine / cinnamic acid ion complex exhibits amphiphilicity and can be self-assembled in the water phase. As a result, the polyethyleneimine / cinnamic acid ion complex is opaque at a low temperature range, and at high temperature, the solubility of cinnamic acid is increased at an elevated temperature, resulting in loss of amphiphilic property of the polyethyleneimine / .

Korean Patent No. 10-1387059 (registered on Apr. 14, 2014) discloses a monoolein cubic phase nanoparticle surface-modified with a hydrophobic cationic polymer, and a method for preparing the same, wherein the hydrophobic modified cationic polymer The monoolein cubic phase nanoparticles are prepared by adding an aqueous solution containing a target material to a monoolein melt and then hydrating to prepare a monoolein cubic phase in which a target substance is collected in a water channel ; The monoolein cubic phase nanoparticles coated with the cationic polymer having the outer surface hydrophobized by adding the hydrophobicized cationic polymer and the water-dispersed aqueous phase to the monoolein cubic phase on which the target substance is captured are added and then homogenized to prepare the monoolein cubic phase nanoparticles coated with the cationic polymer having the hydrophobic outer surface Wherein the outer surface of the monoolein cubic nanoparticles is modified with a hydrophobic cationic polymer to increase skin retention by electrostatic interaction with the skin surface, And thereby improving the permeability of the target substance. Korean Patent No. 10-1055804 (registered on Aug. 23, 2011) relates to a monoolein cubic phase nanoparticle containing an extract of watery extracts effective for atopic dermatitis, Enhances transdermal absorption and improves the healing efficacy of atopic dermatitis.

The present invention provides a new cubic phase capable of releasing and controlling a substance to be loaded according to temperature through immobilization of a specific ion complex on the cubic phase and a manufacturing method thereof.

The present invention provides a temperature responsive ionic complex formed by ion-bonding of polyethyleneimine and cinnamic acid.

The ion complex of the present invention means a state in which polyethylenimine and cinnamic acid are ionically bonded. Preferably, the ionic complex is formed by ion-bonding of an amine group of polyethyleneimine and a carboxyl group of cinnamic acid.

Meanwhile, the present invention provides a method for producing an ion complex exhibiting temperature responsiveness, which comprises mixing polyethyleneimine and cinnamic acid in water. When polyethyleneimine and cinnamic acid are mixed at the water phase, ion complexes exhibiting temperature responsiveness can be produced.

The present invention provides a temperature responsive cubic phase characterized in that an ionic complex of polyethyleneimine and cinnamic acid is fixed on a monoolein cubic phase.

The monoolein (MO) cubic phase forms a bicontinuous cubic phase in the aqueous phase with amphiphiles whose packing parameters are slightly greater than 1 CV Kulkarni, W. Wachter, G. Iglesias-Salto, S. Engelskirchen, S. Ahualli, Phys. Chem. The cubic phase has two mutually intersecting water channels (about 2-3 nm in diameter) surrounded by a mono-olein bilayer (MO bilayer, about 3.5 nm thick) matrix. The cubic phase is optically transparent because it is isotropy, and the content of water on the cubic is 28 to 40%, and is classified into a gyroid cubic phase and a diamond cubic phase depending on the content of water. The cubic phase has a merit that water-soluble compounds and lipid-soluble compounds can be loaded at the same time since the cubic phase includes a water channel and a lipid matrix, and the water-soluble compound can be mounted on the water channel and the lipophilic compound can be mounted on the matrix.

On the other hand, the temperature responsive cubic phase of the present invention means a cubic phase in which a cubic phase matrix is formed of materials having characteristics of being aggregated and dissolved according to temperature, thereby controlling the emission of a target material.

On the other hand, in the temperature responsive cubic phase of the present invention, it is preferable that the ion complex is formed by ionic bonding of an amine group of polyethyleneimine and a carboxyl group of cinnamic acid.

The present invention also provides a process for producing a temperature responsive cubic phase, characterized in that Cinnamic Acid and Monoolein are mixed in an aqueous phase and then polyethyleneimine is added. A temperature responsive cubic phase can be prepared by mixing Cinnamic Acid and Monoolein with water and then adding polyethyleneimine.

The temperature-responsive cubic phase of the present invention is characterized in that the polyethyleneimine / cinnamic acid ion complex, which is characterized by self-assembling and self-disassembling depending on the temperature change, And exhibits an effect of emitting in response to a temperature change.

1 is an illustration of a monoolein cubic phase containing a polyethyleneimine / cinnamic acid ion complex.
FIG. 2 is a graph showing the temperature dependent transmittance of the polyethyleneimine / cinnamic acid mixed solution of each molar ratio in the heating process () and the cooling process ().
3 is a graph showing the temperature dependent transmittance of the polyethyleneimine / cinnamic acid mixed solution (2xd) of each molar ratio in the heating process () and the cooling process ().
4 is a graph showing the temperature dependent permeation of the polyethyleneimine / cinnamic acid mixed solution (4xd) of each molar ratio in the heating process () and the cooling process ().
FIG. 5 is a graph showing the results of measurement of monoolein cubic phase (0/0), (0/10), (0/15), (0/20), (0 / 50) and (0/100).
6 is a graph showing the phase transition temperature of monoolein cubic phase according to the content of cinnamic acid in FIG.
FIG. 7 shows the results of measurement of a monoolein cubic phase (0/0) (A) and a monoolein cubic phase (2.9 / 15) at 25 ° C. (), 30 ° C. (), 45 ° C. ) ≪ / RTI > (B).
8 is an optical microscope photograph of a solution of polyethyleneimine / cinnamic acid (Preparation Example 6, 2xd) observed at 20 占 폚 (A) and 50 占 폚 (B) during the heating process.

In the present invention, a temperature responsive cubic phase was developed by incorporating cinnamic acid and polyethyleneimine into monoolein cubic phase (see FIG. 1). 1 is an illustration of a monoolein cubic phase containing a polyethyleneimine / cinnamic acid ion complex.

The amine group of polyethyleneimine and the carboxyl group of cinnamic acid electrostatically interact to form an ion complex. The polyethyleneimine / cinnamic acid ion complex self-assembles into gel particles in the aqueous phase and exhibits the upper critical solution temperature (UCST) behavior. That is, when the temperature is below the phase transition temperature (UCST), the polyethyleneimine / cinnamic acid ion complex is self-aggregated, thereby suppressing the diffusion of the solute through the water channel of the cubic phase. Conversely, Is disassembled to facilitate the diffusion of the solute in the water channel.

Meanwhile, in the present invention, the phase transition temperature (USCT) behavior was measured by diluting the Shin-Nansan / polyethylene imine solution 2-fold and 4-fold, respectively. When the solution was diluted, it was confirmed that the UCST behavior disappears in the solution exhibiting the UCST behavior before the dilution, because the dilution causes the long-distance hydrophobic interaction between the mutant molecules to be disadvantageous, The intermolecular interaction of the polyethyleneimine / cinnamic acid ion complex is lowered, and it is difficult to cause aggregation. That is, the concentration of the polyethyleneimine / cinnamic acid ion complex is lower than the critical aggregation concentration due to the dilution, and the UCST behavior has disappeared.

On the other hand, contrary to the above results, it was confirmed that the UCST behavior after dilution was observed in the solution in which the UCST behavior before dilution was not observed. In such cases, the number of cinnamic acids bound per molecule of polyethyleneimine in the solution before dilution is large. In this case, the UCST behavior should not appear because the polyethyleneimine / cinnamic acid ion complex still exhibits amphiphilic properties and self-aggregation even when heated to a high temperature. However, when the amount of cinnamic acid bound to a polyethyleneimine molecule is decreased due to the dilution of the solution, the distance between the molecules of the cinnamic acid is increased, and the molecules of the cinnamic acid are solubilized to show the UCST behavior. In other words, by dilution, the hydrophobic mutual attraction between Shin-Na acid molecules is reduced, and the intermolecular attraction between the polyethyleneimine / cinnamic acid ion complex is reduced, and the UCST behavior appears.

In the present invention, the polyethyleneimine / cinnamic acid solution exhibits a reversible UCST behavior in the temperature range of 20 to 80 ° C, and the concentration of the polyethyleneimine / cinnamic acid ion complex and the ratio of polyethyleneimine and cinnamic acid determine the temperature dependency It can be seen that That is, the requirement for the polyethyleneimine / cinnamic acid ion complex to exhibit the UCST behavior is that the polyethyleneimine / cinnamic acid ion complex should self-aggregate at low temperatures.

In order to meet this requirement, the amphiphilicity of the polyethyleneimine and cinnamic acid ion complexes must be high enough at low temperatures. Since amphipathicity depends heavily on the hydrophilic / lipophilic balance value, the ratio of polyethyleneimine and cinnamic acid is an important variable in determining the occurrence of UCST behavior.

Also, for self-aggregation at low temperatures, the concentration of the polyethyleneimine / cinnamic acid ion complex should be higher than the critical aggregation concentration. For this reason, the concentration of polyethyleneimine and cinnamic acid (dilution factor) is another important variable in determining the UCST behavior of the solution.

As a result, in the present invention, Shin-Na acid is inserted into the lipid bilayer on cubic to orient the carboxyl group of Shinnam acid to the water-filled microchannel, and the polyethyleneimine is bonded to Shinnam acid by the electrostatic attraction of its amino group and the carboxyl group of Shin- Ion complex to form a water channel. The polyethyleneimine contained in this water channel acts as a resistance to dye diffusion and determines the release of the substance loaded on the cubic according to the temperature.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the scope of the present invention is not limited to the following embodiments, and includes modifications of equivalent technical ideas.

[ Example  1: polyethyleneimine ( Polyethyleneimine ) / Shinnam-san  ( Cinnamic  acid) ion complexes of UCST]

In this example, the effect of the temperature change on the polyethyleneimine / cinnamic acid ion complex was examined.

(1) Production of polyethyleneimine / cinnamic acid ion complex

Polyethyleneimine and cinnamic acid were dissolved together in 5 ml of a Trizma bases solution (pH 7.0) to prepare a mixture solution containing an ion complex. 8: 2, 7: 3, 6: 4, 5: 5, 4: 6, 3: 7, 2: 8 and 1: 9, and the sum of the molar concentrations of amino group and carboxyl group was kept constant. Preparation Examples 1 to 9 under the above conditions were prepared in accordance with the capacity of the following Table 1.

Production Example 1 Production Example 2 Production Example 3 Production Example 4 Production Example 5 Production Example 6 Production Example 7 Production Example 8 Production Example 9 The molar ratio (PEI: CA) 9: 1 8: 2 7: 3 6: 4 5: 5 4: 6 3: 7 2: 8 1: 9 Polyethyleneimine (PEI)
(Unit: mg / ml) 13.2 11.8 10.3 8.8 7.4 5.9 4.4 2.9 1.5 Shinnam-san (CA)
(Unit: mg / ml) 5 10 15 20 25 30 35 40 45

(2) Polyethylene imine / Shin-Nam-san  According to the addition ratio ( Manufacturing example  1 to 9) UCST  Check the behavior

The temperature-dependent transmittance of the polyethyleneimine / cinnamic acid mixture solution prepared above was measured using a UV spectrophotometer (JENWAY 6505, UK) at a temperature range of 20 to 80 캜, and the polyethylene- The optical density of the imine / cinnamic acid mixture solution was measured and confirmed.

Production Examples 1 to 9 prepared in Example 1 were preheated at 80 DEG C and then cooled at a rate of 5 DEG C / min. Conversely, each of Production Examples 1 to 9 was heated to 80 DEG C at the same rate, and the optical density of each production example was measured during the heating and cooling process.

As a result of the experiment, it was confirmed that the molar ratio of polyethyleneimine and cinnamic acid was 7: 3, that of Preparation Example 1 in which the molar ratio of polyethyleneimine and cinnamic acid was 9: 1, that in Preparation Example 2 in which the molar ratio of polyethyleneimine and cinnamic acid was 8: Of Production Example 3 was totally transparent at 20 to 80 캜 which is the temperature range of the experiment ((A) to (C) of FIG. 2). This means that Production Examples 1 to 3 did not show UCST behavior in the temperature range of 20 to 80 캜, which indicates that the amount of cinnamic acid was not sufficient to allow the polyethyleneimine and cinnamic acid ion complexes to have amphiphilic properties .

On the other hand, Example 4, in which the molar ratio of polyethyleneimine and cinnamic acid was 6: 4, was transparent when cooled from 80 to 75 DEG C, and transparency decreased sharply when cooled to 75 to 60 DEG C, And was opaque at a temperature range of 25 캜 (FIG. 2 (D)).

From the above experimental results, it can be confirmed that the preparation example 4 shows a reversible upper critical solution temperature (UCST) behavior in the water phase.

On the other hand, the temperature-dependent transmittance profile obtained in the heating process of Production Examples 1 to 4 was almost the same as the transmittance profile obtained in the cooling process.

Meanwhile, Production Example 5 showed reversible UCST behavior as in Production Example 4, and the temperature range in which the transmittance change occurred was almost the same as the temperature range in which the transmittance of Production Example 4 was changed (see FIG. 2 (E)).

On the other hand, Production Examples 6 to 9 were opaque at all temperatures and did not show UCST behavior. This is because the lower the molar ratio of the amino group to the carboxyl group (i.e., the greater the relative amount of cinnamic acid relative to polyethylene), the greater the number of cinnamic acid bound to one molecule of polyethylene, the greater the amount of cinnamic acid, (Fig. 2 (F) to (I)). As shown in Figs. 2 (F) to 2 (I), the polyethyleneimine and cinnamic acid ion complexes were still amphipathic.

FIG. 2 is a graph showing the temperature dependent transmittance of the polyethyleneimine / cinnamic acid mixed solution of each molar ratio in the heating process () and the cooling process ().

(3) Polyethylene imine / Shinnam-san  The concentration of the mixture solution UCST  Identify impact on behavior

In order to examine the influence of the concentration of the polyethyleneimine / cinnamic acid mixture solution on the temperature dependency, the optical density of the diluted solutions was measured by diluting the optical density of the diluted solutions by 2 and 4 times, respectively, in Production Examples 1 to 9 prepared in Example 1 . ≪ / RTI > Production Examples 1 to 9 diluted by 2 times and Production Examples 1 to 9 diluted by 4 times are represented by "(2xd)" and "(4xd)", respectively.

As a result of the test, the products of Production Example 1 (2xd), Production Example 2 (2xd), Production Example 3 (2xd) and Production Example 4 (2xd) were transparent at all temperatures ((A) to (D) of Fig. According to the results of the above tests, Production Example 4, which was not diluted, exhibited UCST behavior (FIG. 2 (D)), but Production Example 4 (2 × d) which was prepared by diluting Production Example 4 by 2 fold did not show UCST behavior.

In addition, although the permeability of Production Example 5 (2xd) was somewhat reduced as it was cooled from 45 占 폚 to 25 占 폚, the UCST behavior was not observed clearly (Fig. 3 (E)). The undiluted Preparation Example 5 showed a distinct UCST behavior (Fig. 2 (E)).

The reason for the disappearance of the UCST behavior due to the dilution is that the intermolecular hydrophobic interaction of the polyethylene / cinnamic acid ion complex becomes disadvantageous as the polyethyleneimine / cinnamic acid mixture solution is diluted, and the polyethyleneimine / And that it was difficult for self-aggregation of cinnamic acid ion complexes to lower the critical aggregation concentration.

On the other hand, in Production Examples 4 to 5, UCST behavior was observed by dilution in Production Examples 6 to 9 in contrast to the disappearance of UCST behavior due to dilution. That is, the undiluted Production Examples 6 to 9 were opaque at the entire temperature range tested without showing the UCST behavior ((F) to (I) in FIG. 2) (FIG. 3 (F)), it was confirmed that the UCST behavior was also observed in Production Examples 7 to 9 (2xd) which were diluted twice (see FIG. 3 (G) to (I)).

This means that when the preparation examples 6 to 9 are diluted, the intermolecular attraction between the polyethyleneimine / cinnamic acid ion conjugates is reduced and self-aggregation can be achieved at a lower temperature range, and the polyethyleneimine / It was judged to be temperature-sensitive. 3 is a graph showing the temperature dependent transmittance of the polyethyleneimine / cinnamic acid mixed solution (2xd) of each molar ratio in the heating process () and the cooling process ().

The results of the above dilution experiments show that the UCST behavior characteristics of the polyethyleneimine / cinnamic acid ion complex are as follows: Production Example 9 (2xd) Production Example 8 (2xd) Production Example 7 (2xd) Production Example 6 (2xd) Respectively. This is because when the amount of cinnamic acid relative to the amount of polyethyleneimine is relatively large, the number of cinnamic acid molecules bonded to the polyethyleneimine molecule increases, and the intermolecular hydrophobic interaction formed between the polyethyleneimine / It was concluded that the self - assembly of polyethyleneimine / cinnamic acid complexes was less stable for heat and less sensitive to UCST behavior due to the hydrophobic interaction between the molecules.

On the other hand, according to the results of the temperature dependency transmittance test of the production example (2xd) diluted twice as described above, the production examples 6 and 7 (2xd) exhibited the UCST behavior ((F) and (G) Examples 6 and 7 (4xd) appeared transparent at all temperature conditions and did not show UCST behavior (Fig. 4 (F) and (G)). That is, Production Examples 6 and 7 (2xd) exhibiting the UCST behavior disappeared due to a 2-fold dilution.

On the other hand, the permeability of the solution of Preparation Example 8 (4xd) diluted 4-fold was slightly decreased as the temperature was decreased from 35 占 폚 to 20 占 폚, but the UCST behavior was not apparent (FIG. It is confirmed from the previous experiment result that the 2xd dilution of Preparation Example 8 (2xd) showed distinct UCST behavior (FIG. 3 (H)), and the UCST behavior of Production Example 8 (4xd) disappears due to dilution. This is because the dilution reduces the intermolecular hydrophobic attraction of the polyethyleneimine and cinnamic acid ion complexes, and the self-assembly of the polyethyleneimine / cinnamic acid ion complex is hardly formed at a low temperature of 20 ° C.

On the other hand, Production Example 9 (2xd) did not show a clear UCST behavior (Fig. 3 (I)), but the 2x-diluted preparation of Example 9 (4xd) solution exhibited reversible UCST behavior (Fig. 4 (I)). This is because the intensities of the intermolecular attraction between the polyethyleneimine / cinnamic acid ion complexes were moderately reduced by dilution, and the polyethyleneimine / cinnamic acid ion complexes exhibited temperature dependency at all temperature conditions. 4 is a graph showing the temperature dependent permeation of the polyethyleneimine / cinnamic acid mixed solution (4xd) of each molar ratio in the heating process () and the cooling process ().

In summary, the reversible UCST behavior in polyethyleneimine / cinnamic acid mixture solution (pH 7.0) was found to vary in the temperature range of 20 to 80 ° C. depending on the degree of dilution, / Shinnum acid ion complex and the ratio of polyethyleneimine and cinnamic acid are important parameters for determining temperature dependence.

Example 2 Preparation of Monoolein (MO) Cubic Phase Containing Polyethyleneimine / Cinnamic acid Ion Complex and Determination of Phase Transition Temperature

In this example, a monoolein cubic phase containing a polyethyleneimine / cinnamic acid ion complex was prepared and the influence of the polyethyleneimine / cinnamic acid ion complex on the temperature dependent emission characteristics of the cubic phase was investigated.

(1) Production of monoolein cubic phase containing polyethylenemine / cinnamic acid ion complex

2.3 g of monoolein and varying amounts of cinnamic acid (0, 10, 15, 20, 50 and 100 mg) were placed in a 20 ml glass bottle and the mixture was melted in a water bath at 50 ° C in a water bath. 1 ml of tri-HCl buffer (30 mM, pH 7.0) preheated to the same temperature was carefully added to the top of the mixed melt and sealed with a lid until the buffer solution was completely absorbed and a transparent gel was formed. And stored in a cow. Then, 2.9 mg of polyethyleneimine and 1 mg of methylene blue solution were dissolved in tri-HCl buffer (30 mM, pH 7.0) in order to observe the effect of the polyethyleneimine / cinnamic acid ion complex on the temperature dependent release characteristics of monoolein cubic phase. 1 ml of a mixed solution of polyethyleneimine and methylene blue was used to hydrate a mixed melt of monoolein (2.3 g) and cinnamic acid (15 mg) in place of 1 ml of tri-HCl buffer. Using the above production conditions, the 'concentration of polyethyleneimine and cinnamic acid' on the cubic phase and the molar ratio of 'amino group / carboxyl group' containing polyethyleneimine and cinnamic acid were the same as those of the polyethyleneimine And " concentration of cinnamic acid " and " molar ratio of amino group / carboxyl group ". (The solution of Preparation Example 6 (2xd) prepared in Example 1 was confirmed to exhibit the UCST behavior in the temperature range of 32 to 50 占 폚).

On the other hand, a monoolein cubic phase containing polyethyleneimine (a / mg) and cinnamic acid (b / mg) was variously prepared and named monoolein cubic phase (a / b). As a control sample, 2.3 g of the monoolein melt was hydrated with 1 ml of methylene blue solution to obtain a monoolein cubic phase (0/0) containing no cinnamic acid and polyethyleneimine, and a monoolein cubic phase (0/10), (0/15), (0/20), (0/50) and (0/100) were prepared and used in the following experiments.

(2) Determination of the phase transition temperature of monoolein (MO) cubic phase containing polyethylenimine / cinnamic acid ion complex

The phase transition temperatures of the polyethyleneimine and monoolein cubic phase containing cinnamic acid prepared above were observed with a polarizing microscope (CX31, OLYMPUS). A 'poly (ethylene terephthalate) film' in the form of an O-ring having a thickness of 100 μm was fixed to a cover glass and used as a cell for holding a monoolein cubic phase. The cell containing the monoolein cubic phase was filled in the O-ring space of the cell and covered with another cover glass, and the cell containing the monoolein cubic phase was placed in a heating device (HS1, Mettler Toledo) equipped with a thermostat (Hot Stage (HS82, Mettler Toledo)) and then heated at a rate of 2 占 폚 / min in a temperature range of 25 to 80 占 폚. Then, the cubic texture was observed with a polarizing microscope.

As a result of the experiment, no pattern was observed in the temperature range of 25 to 60 占 폚 for the monoolein cubic phase (0/0) and phase transition was observed at 60.5 占 폚 (FIG. 5 (A)). Generally, since the cubic phase exhibits optically isotropic properties, polarized light is not birefringent when passing through the cubic phase. However, in the case of the monoolein cubic phase (0/0), birefringence was observed at a temperature of 60.5 ° C or higher. It was judged that the monoolein cubic phase underwent phase transition to a reversed hexagonal phase at a specific temperature and that the polarized light was observed to be birefringent when passing through a reversed phase hexagonal phase (JC Shah, Y. et al. Sadhale, DM Chilukuri, Adv. Drug Delivery Rev. 47 (2001) 229.).

On the other hand, the monoolein cubic phase (0/10) showed phase transition at 59 DEG C, 1.5 DEG C lower than the phase transition temperature of the monoolein cubic phase (0/0) (Fig. 5B) The phase transition temperatures of cubic phase (0/0), (0/10), (0/15), (0/20), (0/50) and (0/100) were 60.5 ℃, 59 ℃, 58.5 ℃ , 56.5 캜, 55.5 캜 and 53.5 캜 (Fig. 5 (C) to (F)). As a result, it was confirmed that the phase transition temperature of the monoolein cubic phase was decreased as the content of cinnamic acid was increased (FIG. 6).

This is because the phenyl group of the hydrophilic Shinnam acid is inserted into the monoolein bilayer membrane and the hydrophilic carboxyl group is directed to the water channel and ShinNam acid is inserted into the monoolein bilayer and the lipid bilayer membrane is fluidized to decrease the phase transition temperature .

FIG. 5 is a graph showing the results of measurement of monoolein cubic phase (0/0), (0/10), (0/15), (0/20), (0 / 50) and (0/100).

6 is a graph showing the phase transition temperature of monoolein cubic phase according to the content of cinnamic acid in FIG.

[ Example  3: Confirmation of temperature dependency release]

In this example, the degree of the release of the substance (methylene blue) loaded on the monoolein cubic phase containing Shin-Nam-san and polyethyleneimine was examined according to the temperature.

4 ml of Trigmase Base (30 mM, pH 7.0) as a release medium was carefully added onto the monoolein cubic phase (2.6 / 15) in a 20 ml glass bottle, tightly sealed with a cap, The vial containing the release medium was shaken briefly while being stored at 25 캜, 30 캜, 45 캜 and 50 캜, respectively. After that, 0.1 ml of the release medium was taken out from the glass bottle at each temperature, and the amount of methylene blue released after 0, 3, 6, 12, 24, 48 and 72 hours was quantified. At this time, the release medium was collected and the volume of the release medium was kept constant by supplementing the amount of the reduced release medium by adding the same amount (0.1 ml) of buffer solution to the vial. The amount of methylene blue released from the monoolein cubic phase was determined by measuring the absorbance at 610 nm using a UV spectrophotometer (JENWAY 6505, UK).

Generally, reservoir type of drug carrier has a zero-order release behavior and matrix type of drug carrier has a primary release behavior (J.-C. Kim, Lee, J.-D. Kim, Colloids Surf., B: Biointerfaces 36 (2004) 161, TG. Kim, YK Kim, G. Tae, KY Lee, Park, Biomaterials 20 (1999) 517.). As a result, the release rate of methylene blue in the monoolein cubic phase (0/0) rapidly increased at an early stage, then gradually increased with time, There was almost no change, The release at each temperature condition was similar to the 1st order release. The maximum degree of release obtained at 25, 30, 45 and 50 ° C was 14.7%, 15.5%, 15.2% and 16.1%, respectively, and the maximum drug release degree was not significantly different according to the release temperature (Fig.

On the other hand, since the fine water channel that can contain a water-soluble compound is uniformly distributed in the matrix of cubic, the cubic phase is a matrix-type drug carrier and the diffusion coefficient of the solute in the solution medium is proportional to the absolute square root of the temperature (J. Siepmann, NA Peppas, Adv. Drug Delivery Rev. 48 (2001) 139.) The release rate is faster as the temperature is higher. However, in the present invention, since the width of the temperature change is as small as 25 占 폚, it was judged that the temperature change hardly affected the degree of emission.

On the other hand, the release pattern of methylene blue emitted from the monoolein cubic phase (2.9 / 15) was similar to the release profile of the monoolein cubic phase (0/0) (FIG. 7 (B)). However, the maximum degree of release observed at 25, 30, 45 and 50 ° C was lower than that of the monoolein cubic phase (0/0) at all the temperatures tested at 4.1%, 4.3%, 12.5% and 13.1% .

The phase transition temperatures of the monoolein cubic phases (0/0) and (0/15) determined in Example 2 were 60.5 ° C and 58.5 ° C, respectively (FIGS. 5A and 5C) 30, 45 and 50 ° C, which is the temperature of the emission experiment conducted in the example. Therefore, in Example 3, the reason why the degree of release of the cubic phase (2.9 / 15) is lower than the degree of release of the cubic phase (0/0) is excluded from whether or not the phase transition of the monoolein cubic phase is present. As a result, the monoolein cubic phase (2.9 / 15) forms a polyethyleneimine / cinnamic acid ion complex, which is less than the monoolein cubic phase (0/0) because the polyethyleneimine contained in the water channel inhibits dye diffusion It was judged that a positive dye was released.

On the other hand, according to the emission test results of the monoolein cubic phase (2.9 / 15), the degree of release of the monoolein cubic phase (2.9 / 15) observed at relatively high temperatures (45 and 50 ° C) And 30 [deg.] C) was about three times higher than that of the monoolein cubic phase (Fig. 7 (B)). It was judged that this was due to the self - aggregation of polyethyleneimine and cinnamic acid ion complex. FIG. 7 shows the results of measurement of a monoolein cubic phase (0/0) (A) and a monoolein cubic phase (2.9 / 15) at 25 ° C. (), 30 ° C. (), 45 ° C. ) ≪ / RTI > (B).

On the other hand, the monoolein cubic phase (2.9 / 15) in the above experiment was prepared by hydrating a monoolein melt with an aqueous solution (polyethyleneimine 2.9 mg / ml, Shinnam acid 15 mg / ml) And the molar ratio of the amino group / carboxyl group to the concentration of polyethyleneimine and cinnamic acid in the solution of preparation example 6 (2xd) are the same as the molar ratio of amino group / carboxyl group.

Therefore, it was confirmed that the UCST behavior of the polyethyleneimine / cinnamic acid complex shown in Production Example 6 (2xd) (indicating UCST at around 32 ° C) can also occur on the cubic phase water channel. At temperatures above UCST (for example, 45 and 50 ° C), the polyethyleneimine and cinnamic acid ionic complexes are difficult to self-aggregate and effectively block the cubic phase water channels, but at temperatures below UCST (for example 25 and 30 ° C) , The polyethyleneimine / cinnamic acid ionic conjugate is self-aggregated to effectively block the water channel, thereby suppressing drug release.

[Example 4: Confirmation of temperature dependency of polyethyleneimine / cinnamic acid ion complex and disassembly]

In this example, it was further confirmed that the polyethyleneimine / cinnamic acid ion complex was aggregated and disassembled by temperature.

Production Example 6 (2xd) solution was filled in a rubber O-ring (diameter: 1.0 cm, thickness: 0.2 cm) immobilized on the cover glass surface and covered with another cover glass. Then, the cell containing the polyethyleneimine / cinnamic acid mixture solution was mounted on a heating device (hot stage, HS82, Mettler Toledo) equipped with a thermostat (HS1, Mettler Toledo) At a rate of 2 [deg.] C / min. Thereafter, a photograph of the solution of Production Example 6 (2xd) was observed with an optical microscope (CX31, OLYMPUS, Japan) at a temperature range of 20 占 폚 and 50 占 폚.

As a result of the test, microparticles of several tens of micrometers were observed in the photograph of the mixture solution of Preparation Example 6 (2xd) at 20 ° C. This is the result of confirming that the polyethyleneimine / cinnamic acid ion complex is self-aggregating in the water phase. According to FIG. 3F, Production Example 6 (2xd) was opaque at 20 ° C, which was judged to be due to the microparticles formed by the self-assembly as confirmed in this Example (FIG. 8A).

On the other hand, it was confirmed that microparticles disappeared when heated at 50 DEG C in Production Example 6 (2xd) (Fig. 8B). This is in agreement with the result (F in FIG. 3) that Production Example 6 (2xd) was transparent at 50 ° C as confirmed in Example 1 above.

As a result, the hydrophilicity / hydrophobicity balance (HLB value) of the polyethyleneimine / cinnamic acid ion complex is proportional to the temperature increase, and at a temperature as high as 50 DEG C, the polyethyleneimine / cinnamic acid ion complex loses amphiphilicity, It was confirmed that the particles disappeared.

In summary, the polyethyleneimine / cinnamic acid ion complex is self-assembled and disassembled in a temperature-dependent manner in the aqueous channel of the monoolein-cubic phase. Due to this temperature-dependent self-aggregation phenomenon, a monoolein containing a polyethyleneimine / It was confirmed that the cubic phase had temperature-dependent controlled release characteristics (Fig. 7 (B)).

8 is an optical microscope photograph of a solution of polyethyleneimine / cinnamic acid (Preparation Example 6, 2xd) observed at 20 占 폚 (A) and 50 占 폚 (B) during the heating process.

Claims (6)

Polyethyleneimine and Cinnamic Acid are formed by ionic bonding,
The concentration of polyethyleneimine is 2.9 mg / ml, the concentration of cinnamic acid is 15 mg / ml,
Wherein the molar ratio of the amino group of the polyethyleneimine to the carboxyl group of the cinnamic acid is 4: 6.
The method according to claim 1,
In the ion complex,
Characterized in that an amine group of polyethylenimine and a carboxyl group of cinnamic acid are ionically bonded to form a temperature responsive ion complex.
Polyethyleneimine and Cinnamic Acid are blended at the awards,
The concentration of polyethyleneimine is 2.9 mg / ml, the concentration of cinnamic acid is 15 mg / ml,
Wherein the molar ratio of the amino group of the polyethyleneimine to the carboxyl group of the cinnamic acid is 4: 6.
A temperature responsive ionic complex formed by the ionic bonding of polyethyleneimine and cinnamic acid is fixed on the monoolein cubic phase,
The concentration of polyethyleneimine is 2.9 mg / ml, the concentration of cinnamic acid is 15 mg / ml,
Wherein the molar ratio of the amino group of the polyethyleneimine to the carboxyl group of the cinnamic acid is 4: 6.
5. The method of claim 4,
In the ion complex,
A temperature responsive cubic phase characterized in that an amine group of polyethyleneimine and a carboxyl group of cinnamic acid are formed by ionic bonding.
Cinnamic Acid and Monoolein are mixed in water phase, then polyethyleneimine is added,
The concentration of polyethyleneimine is 2.9 mg / ml, the concentration of cinnamic acid is 15 mg / ml,
Wherein the molar ratio of the amino group of the polyethyleneimine to the carboxyl group of the cinnamic acid is 4: 6.

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