KR100768151B1 - Nanoliposome using esterified lecithin - Google Patents

Abstract

A nano-liposome using esterified lecithin is provided to maintain the long term stability and uniformity, thereby being used for cosmetics with improved moisturizing capability and permeation capability. A nano-liposome comprises a liposome membrane including esterified lecithin which is a reaction product of hydrogenated saturated hydrocarbon lecithin and an organic acid and a physiologically active effective ingredient encapsulated into the liposome membrane, wherein the physiologically active effective ingredient is coenzyme Q10 or an epidermal growth factor(EGF) and the organic acid is selected from the group consisting of acetic anhydride, malic anhydride, lactic anhydride, glycolic anhydride, citric anhydride, and oxalic anhydride. A method for preparing the nano-liposome comprises the steps of: (a) reacting hydrogenated saturated hydrocarbon lecithin with an organic acid to prepare esterified lecithin; (b) dissolving the esterified lecithin and a physiologically active effective ingredient in a solvent; and (c) dispersing the solution prepared from the step (b) into nano-sized liposome.

Description

Nanoliposomes using esterified lecidine {NANOLIPOSOME USING ESTERIFIED LECITHIN}

Figure 1 is a graph showing the ultraviolet / visible light absorbance (%) of the epithelial cell growth factor / coenzyme Q10 bilayer nanoliposome solution prepared using the esterified lecidine according to the present invention.

Figure 2 is a graph showing the UV / visible light transmittance (%) of the epithelial growth factor / coenzyme Q10 bilayer nanoliposome solution prepared using the esterified lecidine according to the present invention.

Figure 3 is a graph showing the particle distribution of the epithelial growth factor / coenzyme Q10 bilayer nanoliposomes prepared using the esterified lecidine according to the present invention.

The present invention relates to liposome membranes comprising esterified lecidine; And it relates to a nano-liposomes comprising the biologically active ingredient contained in the liposome membrane and a method for producing the same.

Coenzyme Q10 was originally known to have a strong antioxidant against free radicals. Therefore, when ingested or applied to the skin to prevent the oxidation of the cells to maintain the skin elasticity, it is possible to effectively prevent aging. In particular, the coenzyme Q10 is necessary enough to produce until the age of 20, but due to various factors such as unbalance or stress decreases to around 20 years as a peak, and the reduction is accelerated around 40 years old, it is necessary to supplement this.

Meanwhile, epidermal growth factor (EGF) is an excellent protein for promoting cell regeneration and wound healing in colostrum of breast milk, and is a biopharmaceutical used to treat foot ulcers in diabetic patients. In addition, EGF is known to have a natural rejuvenation effect on wounds without scars. Therefore, it is not only used for the healing of wounds such as diabetic foot ulcers, burns and wounds, but also widely used as a functional cosmetic raw material.

Liposomes are microvesicles with a hydrophilic space inside and a closed double lipid membrane outside. Therefore, liposomes are characterized by being able to contain a water-soluble substance in a central hydrophilic space and to collect an oil-soluble substance in an external lipid bilayer. The liposome-forming substance is called lipoid, and lipoid is most commonly used in cosmetics or foods because it has excellent moisturizing properties and is harmless to humans.

However, since these lecidines and ceramides are hydrophobic and hardly dispersed in an aqueous solution as well as an alcoholic solution, in order to use these compounds as lipoids, it is necessary to add them after heating and dispersing them at a high temperature of 70 ° C. or higher. Therefore, there is a very big problem in liposomes of functional compounds oxidized or thermally unstable at high temperatures such as coenzyme Q10 and EGF. In addition, the liposomes thus formed have very low stability, and the size of the liposomes is not uniform. In particular, it is difficult to liposome two functional substances with greatly different polarities, such as hydrophobic coenzyme Q10 and hydrophilic EGF.

Recently, phospholipid lipoids in the form of anionic surfactants having good dispersibility in aqueous solutions by reacting lecidin with phosphoric acid or other polar compounds are used. However, if the lipoid is too water soluble, the wettability is good, but the skin penetration effect is greatly reduced due to the polarity difference between the skin surface components, and if the lipoid is in the form of an anion salt, the viscosity of the cosmetic should be further added to further increase the thickener. have.

Objects of the invention are liposome membranes comprising esterified lecidine; And it is to provide a nano liposome comprising a physiologically active ingredient contained in the liposome membrane. According to the present invention, when a double membrane of liposomes is formed by using esterified lecidin having good dispersibility and hydrophobicity in both water and alcohol, nanoparticles containing coenzyme Q10 or epidermal growth factor as a functional substance The liposome solution can be prepared even at low temperature, and the nano liposome thus prepared can be used as a raw material for producing cosmetics having excellent moisture and permeability because it is stable and uniform for a long time.

It is another object of the present invention to provide a method for preparing a stable nanoliposome by dispersing a mixed solution of esterified lecidine and a bioactive active ingredient to a nano size.

The present invention relates to liposome membranes comprising esterified lecidine; And it relates to a nano-liposomes comprising a physiologically active ingredient contained in the liposome membrane.

In addition, the present invention,

A first step of reacting lecidine with an organic acid to produce esterified lecidine;

A second step of dissolving the esterified lecidine and the bioactive active ingredient in a solvent; And

The third step of dispersing the solution prepared in the nano-sized liposomes

It relates to a method for producing a nanoliposome comprising a.

The esterified lecidine used in the present invention has a polarity enough to be dispersed in a water-soluble solvent, but does not show a large polarity enough to completely dissolve in a water-soluble solvent. There is a property to prepare a stable liposome solution in.

The esterified lecidine in the present invention may be prepared by reacting lecidine with an organic acid. The esterification process is a condensation reaction in which the alcohol group of the lecidine reacts with the organic acid to remove water molecules. The esterified lecidine is dissociated again in the form of lecidine (alcohol group) and the organic acid by reacting with water in an aqueous solution. Therefore, this reaction coexists in the form of esterified lecidine, lecidine, and organic acid while maintaining the proper equilibrium of the following dissociation reactions depending on the acidity.

The organic acid used in the preparation of the esterified lecidine of the present invention is preferably an organic acid, and the organic acid may be used in cosmetics and foods such as acetic anhydride, malic anhydride, lactic anhydride, glycolic anhydride, citric anhydride, and oxalic anhydride. Any organic anhydride that can be used is possible, and more preferably malic anhydride or acetic anhydride is used. When acetic anhydride is used, the esterified lecidine exhibits greater hydrophilicity than when malic anhydride is used. Therefore, an organic anhydride can be used as appropriate depending on the relative amount of the hydrophilic or hydrophobic functional material to be added or the degree of polarity of the functional material. These organic anhydrides exhibit exfoliation effects or skin softening effects when applied to the skin.

The bioactive active ingredient entrapped in the nanoliposomal membrane according to the present invention is not limited thereto, but is a water-soluble drug, a fat-soluble drug, or a thermally labile functional material. Preferably, the bioactive active ingredient in the present invention is at least one selected from the group consisting of coenzyme Q10 and epidermal growth factor. At this time, it is preferable to use 1-5 parts by weight of esterified liposomes with respect to 1 part by weight of the bioactive active ingredient in the nanoliposomes.

Nanoliposomes of the present invention are triglyceride organic compounds, preferably capric / capric triglycerides; Anionic surfactants, preferably diethylamine cetylphosphate, ascorbyl phosphate sodium, phosphatidylcholine or triethylamine cocoyl glutamine sodium; Hydrogenated saturated hydrocarbon lecidine; Emollients, preferably butylated hydroxy toluene; Betaine-based amphoteric surfactants, preferably rolyaminepropylbetaine, laurylbetaine, laurylaminebetaine or cocamido propylbetaine; Chelating agents, preferably one or more selected from the group consisting of ethylenediaminetetraacetate sodium salt.

The present invention also provides a method for preparing esterified lecidine by reacting lecidine with an organic acid; A second step of dissolving the esterified lecidine and the bioactive active ingredient in a solvent; And a third step of dispersing the solution prepared above into nano-sized liposomes.

In the first step, esterified lecidine is prepared by condensation of lecidine in an organic acid. Here, the organic acid is preferably selected from the group consisting of acetic anhydride, malic anhydride, lactic anhydride, glycolic anhydride, citric anhydride, and oxalic anhydride.

In the second step, the bioactive active ingredient is not limited thereto, but water-soluble drugs, fat-soluble drugs, or thermally labile functional substances are preferable, and more preferably, one selected from the group consisting of coenzyme Q10 and epidermal growth factor. That's it.

Here, in the case of using coenzyme Q10 and epidermal growth factor as a physiologically active ingredient, preferably, the second step comprises the steps of dissolving esterified lecithin and coenzyme Q10 in an organic solvent to prepare an oily solution; Preparing an aqueous solution by dissolving epithelial growth factor in a solvent; And it may include the step of mixing the oil phase solution and the aqueous solution prepared above.

In the step of preparing the oil phase solution, the esterified lecidine and coenzyme Q10 prepared above are dissolved in an organic solvent such as ethanol to prepare an oil phase solution. Preferably hydrophilic triglyceride organic compounds such as hydrogenated saturated hydrocarbon lecidine, caprylic / capric triglyceride, diethylamine cetylphosphate, ascorbyl phosphate sodium, phosphatidylcholine or triethylamine coco in organic solvents Anionic surfactants such as sodium glutamine sodium or emollients such as butylated hydroxy toluene may be further added.

In the preparation of the aqueous phase solution, the aqueous phase solution is prepared by dissolving EGF, a skin growth promoter, in pure water. Preferably, an aqueous solution is further added by adding an anionic surfactant such as diethylamine cetylphosphate, ascorbyl phosphate sodium, phosphotidylcholine or triethylamine cocoyl glutamine sodium, or a chelating agent such as ethylenediaminetetraacetate sodium salt. Can be prepared.

The process of dissolving the esterified lecidine and the bioactive active ingredient in a solvent is preferably carried out at a temperature of 20 to 60 ℃. At this time, if the temperature is less than 20 ℃ is dissolved, but the time to dissolve is long and even after dissolution is a problem in stability, if it is larger than 60 ℃ bioactive active ingredients coenzyme Q10 and EGF may be unstable, which is not preferable. .

The oil phase solution and the aqueous phase solution prepared above are uniformly stirred using a homomixer stirrer after mixing. At this time, the liposome in the homogeneously mixed liposome solution shows the micrometer size.

In the third step, a nano liposome solution is prepared by dispersing the mixed solution uniformly stirred in the second step into a nano-sized liposome by passing the microfluidizer (M / F) one or more times at a pressure of 1000 psi or more. At this time, if the pressure is less than 1000psi it will not be able to form a nano-sized liposomes can not prepare a preferred nano liposome solution. In addition, the number of passes through the microfluidizer is more effective if treated more than two times.

The liposome of the nanoliposome solution prepared above is able to stabilize the EGF which is the functional material of the innermost aqueous phase by forming double liposomes in the aqueous phase / oil phase, as well as the hydrophilic EGF and the hydrophobic coenzyme Q10. The functional material can be stabilized at the same time.

Coenzyme Q10 and epidermal growth factor-containing nano liposomes using the esterified lecidine according to the present invention can be prepared into cosmetic compositions by conventional methods.

Quantification of the raw materials added in the manufacturing process has been described in detail in the Examples, and according to the Examples. In addition, the additives listed above may be used interchangeably with other additives because of mutual compatibility, and the amount of the additive may be appropriately adjusted according to the amount of the functional material to be manufactured under optimum conditions.

Hereinafter, the present invention will be described in detail with examples. However, the present invention is not limited by the examples.

Example

Synthesis of Esterized Lecidine

<Example 1.>

31 g of hydrogenated saturated hydrocarbon lecidine was dissolved in 200 g of tetrahydrofuran solvent at a temperature of 70 ° C. After the lecidine was completely dissolved, 3 g of malic anhydride was added to dissolve it. After the solution was sufficiently clear, 0.5 g of an alkyl amine or an alcohol amine was added as a catalyst, and the reaction was continued for at least 3 hours while refluxing under cooling. After the reaction was completed, the resultant was dried in vacuo at 45 ° C. to obtain an esterified lecidine as a white powder.

<Example 2.>

Esterized lecidine was synthesized in the same manner as in Example 1 using 2.5 g of acetic anhydride instead of malic anhydride.

Preparation of Nanoliposomal Solution

Example 3.

1) Preparation of Oily Solution

To 110 g of ethanol, 15 g of esterified lecidine, 15 g of hydrogenated saturated hydrocarbon lecidine, 200 g of capric / capric triglyceride (MCT), 10 g of diethylamine cetylphosphate, and 0.5 g of butylated hydroxy toluene (BHT) were added. It was dissolved by stirring at the heating to 40 ℃ and cooled to room temperature to prepare an oil phase solution.

2) Preparation of Aqueous Solution

0.5 g of Na-EDTA, 1 g of ascorbyl phosphate (NAP), and 2.4 g of EGF were dissolved in 650 g of water while stirring at room temperature to prepare an aqueous solution.

3) Mixing oil and water solutions

The aqueous solution prepared above was added to the oily solution, and stirred well for at least 5 minutes with a homomixer to ensure uniform mixing.

4) Dispersion into nano-sized liposomes

The prepared liposome solution was prepared by dispersing the mixed solution into a nano-sized liposome at least once using a microfluidizer at a pressure of 1000 psi or more. At the time of passage, it cooled with cooling water so that it might become temperature below normal temperature.

Example 4.

A nanoliposome solution was prepared in the same manner as in Example 3, except that 30 g of esterified lecidine was added.

Example 5.

A nanoliposome solution was prepared in the same manner as in Example 3 using 10 g of lauryl betaine, a betaine-type amphoteric surfactant, in place of diethylamine cetylphosphate, an anionic surfactant.

Example 6.

A nanoliposome solution was prepared in the same manner as in Example 3 using 50 g of esterified lecidine without using both hydrogenated saturated hydrocarbon lecidine and diethylamine cetylphosphate.

<Example 7.>

1) Preparation of Oily Solution

110 g of ethanol, 15 g of esterified lecidine, 15 g of hydrogenated saturated hydrocarbon lecidine, 200 g of capric / capric triglyceride (MCT), 10 g of diethylamine cetylphosphate, 0.5 g of butylated hydroxy toluene (BHT), and coenzyme Q10 10 g was added thereto, heated to about 40 ° C., dissolved with stirring, and cooled to room temperature to prepare an oily solution.

2) In the same manner as in Example 3, an aqueous phase solution was prepared, the oil phase and the aqueous phase solution were mixed, and then dispersed in nano size to prepare a nanoliposomal solution.

<Example 8.>

A nanoliposome solution was prepared in the same manner as in Example 7 using 30 g of esterified lecidine without using hydrogenated saturated hydrocarbon lecidine.

Example 9.

A nanoliposome solution was prepared in the same manner as in Example 7 using 10 g of laurylamine betaine, a betaine-based amphoteric surfactant instead of diethylamine cetylphosphate.

<Example 10.>

Nanoliposomal solution was prepared in the same manner as in Example 7 using 50 g of esterified lecidine without using both hydrogenated saturated hydrocarbon lecidine and diethylamine cetylphosphate.

Example 11.

1) To 110 g of ethanol, 15 g of esterified lecidine, 15 g of hydrogenated saturated hydrocarbon lecidine, 200 g of capric / capric triglyceride (MCT), 0.5 g of butylated hydroxy toluene, and 10 g of coenzyme Q10 were added and the mixture was heated to about 50 ° C. It was dissolved by heating and stirring, and then cooled to room temperature.

2) 2.4 g of EGF was dissolved in 10 g of distilled water, and this solution was added to the solution prepared in 1) above.

3) Using the homomixer, the solution prepared in 2) was sufficiently stirred for about 10 minutes.

4) The solution was passed through a microfluidizer at a pressure of at least 1000 psi.

5) While stirring with a homomixer, this solution was added to 0.5 g of Na-EDTA and 650 g of water in which 1 g of ascorbyl phosphate was dissolved.

6) After the addition, the mixture was sufficiently stirred for about 10 minutes.

7) The solution was passed through a microfluidizer one or more times to prepare a double nanoliposome solution.

<Example 12.>

A liposome solution was prepared in the same manner as in Example 11, except that 35 g of esterified lecidine was used without using hydrogenated saturated hydrocarbon lecidine.

Example 13.

Nanoliposomal solution was prepared in the same manner as in Example 11 using 50 g of esterified lecidine without using both hydrogenated saturated hydrocarbon lecidine and diethylamine cetylphosphate.

Example 14.

1) To 110 g of ethanol, 1 g of diethylamine cetylphosphate, 200 g of capric / capric triglyceride (MCT), 10 g of esterified lecidine, 0.5 g of butylated hydroxy toluene, and 10 g of coenzyme Q10 were added and stirred vigorously at room temperature. Melting while preparing an oily solution.

2) 2.3 g of EGF was dissolved in 10 g of distilled water, and this solution was added to the solution prepared in 1) above.

3) Using the homomixer, the solution prepared in 2) was sufficiently stirred for about 10 minutes.

4) The solution was passed through a microfluidizer at a pressure of at least 1000 psi.

5) While stirring with a homomixer, this solution was added to 0.5 g of Na-EDTA and 650 g of water in which 1 g of ascorbyl phosphate was dissolved.

6) After the addition, the mixture was sufficiently stirred for about 10 minutes.

7) The solution was passed through a microfluidizer one or more times to prepare a bilayer nanoliposome solution.

<Example 15.>

A nanoliposome solution was prepared in the same manner as in Example 14 using 1 g of ascorbyl phosphate sodium instead of diethylamine cetylphosphate.

<Example 16.>

A nanoliposome solution was prepared in the same manner as in Example 14 using 1 g of triethylamine cocoyl glutamine sodium (MIAMI CT130), an anionic surfactant instead of diethylamine cetylphosphate.

<Example 17.>

Instead of diethylamine cetylphosphate, a nanoliposome solution was prepared in the same manner as in Example 14 using 1 g of a betaine-based amphoteric surfactant, rolliamine propyl betaine.

Comparative Example

A nanoliposome solution was prepared in the same manner as in Example 7 using 30 g of hydrogenated saturated hydrocarbon lecidine without using esterified lecidine.

Test Example

<Stability Test>

Prepared in Example 8 (when using only esterified lecidine), Example 7 (when using esterified lecidine and hydrogenated saturated hydrocarbon lecidine together), and Comparative Example (when using only hydrogenated saturated hydrocarbon lecidine) Table 1 compares the average particle size of one nanoliposome solution and the gelation phenomenon when it is left at room temperature for a long time. In addition, the presence or absence of Ostwald ripening phenomenon, which is one of the mechanisms by which materials are deposited on larger particles due to particle size difference between particles in a solution and grows, was also observed (Ostwald, Z Phys. Chem). (34), 1900, 495-5031).

When the liposome solution is left for a long time, a gelation phenomenon generally occurs due to the flocculation phenomenon. The gelation point is a time when the solution is not dispersed or dissolved again even after the solution is stirred again.

TABLE 1

Liposomal Solution Sample Average particle size of liposomes (nm) When gelation occurs Remarks Example 8 470 25 days Gelation progresses slowly Example 7 750 20 days Ostwald ripening also partially progresses with gelation Comparative example 1300 15th Gelation and Ostwald ripening occur simultaneously, resulting in separation of solvent and liposome layers.

* Test temperature: Room temperature (average 20 ℃)

<Measurement of absorbance and transmittance in the ultraviolet / visible region of the nanoliposomal solution>

Absorbance and transmittance in the ultraviolet / visible region of the nanoliposome solution of the EGF (aqueous) / coenzyme Q10 (oil) bilayer form prepared using only the esterified lecidine according to the present invention prepared in Example 8 Measurements are shown in FIGS. 1 and 2, respectively.

It can be seen from FIGS. 1 and 2 that scattering or absorption was hardly observed in the region between 400 nm and 700 nm wavelengths, which indicates that the nanoliposome solution is very transparent and the size of the liposome is very uniform.

<Measurement of particle distribution>

The particle distribution of the liposomes dispersed in the bilayer nanoliposome solution prepared in Example 8 was measured and shown in FIG. 3.

It can be seen from FIG. 3 that the nanoliposomal particles are distributed in a narrow range, indicating that the nanoliposomal solution is very transparent and the size of the liposome is very uniform.

According to the present invention, by forming a liposome membrane containing esterified lecidine, the thermally labile functional material can be stably liposomed at a low temperature, and it is also possible to simultaneously capture both hydrophilic and hydrophobic functional materials. In addition, since the organic anhydride used to remove the dead skin of the skin is applied to the functional group has the advantage of showing various functional effects such as exfoliation effect or skin softening effect.

Claims (9)

Liposome membranes comprising esterified lecidine, which is a reaction product of hydrogenated saturated hydrocarbon lecidine and an organic acid; And a nanoliposome comprising a physiologically active ingredient contained within the liposome membrane. The nanoliposome according to claim 1, wherein the bioactive active ingredient is at least one selected from the group consisting of coenzyme Q10 and epidermal growth factor. delete The nanoliposome of claim 1 wherein the organic acid is selected from the group consisting of acetic anhydride, malic anhydride, lactic anhydride, glycolic anhydride, citric anhydride, and oxalic anhydride. The nanoliposome according to claim 1, further comprising at least one selected from the group consisting of hydrogenated saturated hydrocarbon lecidine, anionic surfactant, triglyceride organic compound, emollient, chelating agent and betaine amphoteric surfactant. A first step of reacting hydrogenated saturated hydrocarbon lecidine with an organic acid to produce an esterified lecidine; A second step of dissolving the esterified lecidine and the bioactive active ingredient in a solvent; And The third step of dispersing the solution prepared in the nano-sized liposomes Method for producing a nanoliposome comprising a. The method of claim 6, wherein the bioactive active ingredient is at least one selected from the group consisting of coenzyme Q10 and epithelial cell growth factor. The method of claim 7, wherein the second step is Preparing an oily solution by dissolving esterified lecithin and coenzyme Q10 in an organic solvent; Preparing an aqueous solution by dissolving epithelial growth factor in a solvent; And Mixing the oil phase solution and the aqueous solution prepared above. 7. A method according to claim 6, wherein the second step is carried out at a temperature of 20 to 60 ° C.

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