KR101787556B1 - Method for producing of nano-flexible vesicles containing active components, and cosmetical composition using thereof - Google Patents

Abstract

More particularly, the present invention relates to a method for producing a nanoflexible vesicle by capturing an active ingredient, and more particularly, to a method for producing a nanoflexible vesicle by capturing an active ingredient, comprising: 1) adding a solvent to a mixture of an active ingredient, a phospholipid and a surfactant, dissolving the mixture in a high- Preparing a solution; 2) adding distilled water to the sol solution and magnetically stirring to form a gel of a lyotropic liquid crystal; 3) gradually adding distilled water while continuing magnet agitation to the gel and applying a shear force to disperse the gel into vesicle particles in water; 4) treating ultrasonic waves in suspension in which vesicle particles are dispersed; And 5) cooling the suspension treated with the ultrasonic wave to room temperature while stirring the magnet. The present invention also relates to a method for producing a nanoflexible vesicle in which an active ingredient is captured. By using the nano-flexible vesicle production method of the present invention, it is possible to manufacture a vesicle having a size of several tens of nanometers having a flexible membrane, and thus the transdermal absorption of the active ingredient caught in vesicles can be enhanced. In addition, it is possible to manufacture vesicles by a simple process without a solvent evaporation process or a high pressure process, and the efficiency of collecting hydrophilic active ingredients can be significantly increased. The cosmetic formulation containing the nano-flexible beacle prepared using the above method exhibits a slight trophic property, and thus can be used for manufacturing a cosmetic composition having excellent feeling of use.

Description

TECHNICAL FIELD [0001] The present invention relates to a method for producing a nano-flexible beacle containing an active ingredient and a cosmetic composition containing the nano-flexible vesicles,

More particularly, the present invention relates to a method for producing a nano-flexible beacle in which an active ingredient is captured, more specifically, a beacycle which can be flexibly deformed using an active ingredient, a phospholipid, a surfactant and a solvent, The present invention relates to a method for producing a cosmetic composition which improves skin permeation absorption of an active ingredient such as a peptide and thixotropic properties are enhanced when a formulation containing a vesicle is made,

Recently, various kinds of peptides have been newly developed and widely used in pharmaceuticals, cosmetics, etc., and the market of peptides has been rapidly increasing worldwide. Since peptides are synthesized from a combination of amino acids, which are protein components, they are attracting much interest in the cosmetics industry as a biocompatible raw material harmless to human body. Especially, they are currently being applied as anti-aging functional materials for wrinkle improvement. In order for peptides to exhibit the wrinkle-improving effect, the peptide must be absorbed deep into the skin through the horny layer of the skin. Most of the peptides are hydrophilic and have a high molecular weight with at least three amino acids. . In general, a substance having a molecular weight of 500 Da or more is considered difficult to penetrate the skin. Various methods for permeating the peptides to skin have been attempted (Non-Patent Document 1).

A method of chemically modifying a peptide by binding an alkyl group to the peptide in order to lower the hydrophilicity of the peptide and increase lipid solubility has been carried out. However, the peptide still retains a high molecular weight, so that the necessary amount of the peptide does not penetrate the skin. Therefore, attempts to solve the low skin permeation problem of peptides with a formulation have been actively sought. The first is the use of percutaneous penetration enhancers, a method commonly used in the pharmaceutical industry. Examples of skin permeation enhancers include terpene, azone, pyrrolidone, menthol, dimethyl sulfoxide and the like, which weaken the protective function of the skin stratum corneum, Thereby enhancing transmission. However, these skin permeation enhancers cause irritation to the skin and are unsafe, so they can not be used as cosmetic ingredients for long-term daily use. The second method involves collecting peptides in vesicle particles to induce skin permeation. Niosomes and liposomes are representative. Liposomes made from phospholipids, which are biomaterials, are getting more attention than nioxides made from nonionic surfactants. Liposomes, however, are limited in their ability to capture, have low collection efficiency, are not simple to manufacture, and have not been widely utilized due to their ability to penetrate skin, which is below expectations.

Since then, modified liposomes have been developed with a slight modification of the liposomes, called transfersomes and ethosomes, which have greatly improved skin permeation efficacy. B. Godin disclosed ethosomes (Non-Patent Document 2), G. Cevc, J. Guo et al. Reported the skin permeation enhancing effect of peptides using transfersom (Non-Patent Documents 3 and 4 ). Deformable liposomes have proved to have superior skin permeation efficacy compared to liposomes, but still suffer from problems such as collection efficiency, manufacturing cost, stability in formulations, and ethosomes, It is still not applied to cosmetics due to the fact that it is used.

Thus, the inventors of the present invention have made a new method for producing a nano-flexible beacle in which a peptide is captured, and confirmed that the skin permeation absorption effect of the peptide is greatly enhanced by the nano-flexible beacle prepared by the above method.

 A. E. B. Heather, N. Sarika, Proteins and Peptides: Strategies for delivery to and. across the skin, J. of Pharm. Sci., 97 (9), 3591 (2008).  B.Godin, E.Touitou, Ethosomes: new prospects in transdermal delivery, Crit Rev Ther Drug Carrier Syst., 20 (1), 63 (2003).  G. Cevc et al., Ultraflexible vesicles, Transfersomes, an extremely low pore penetration resistance and transport therapeutic amounts of insulin across the intact mammalian skin, Biochim Biophys Acta. 1368 (2), 201 (1998).  J. Guo, Q. Ping, and L. Zhang, Transdermal delivery of insulin in mice by using lecithin vesicles as a carrier, Drug Delivery, 7 (2), 113 (2000).

It is an object of the present invention to provide a method for producing a nanoflexible vesicle in which an active ingredient is captured.

Another object of the present invention is to provide a cosmetic composition containing a nano-flexible beacle in which an active ingredient produced by the above method is collected.

Hereinafter, the present invention will be described in detail.

One embodiment of the present invention relates to a method for producing nano-flexible vesicles in which an active ingredient is captured, more specifically,

1) adding a solvent to a mixture of an active ingredient, a phospholipid and a surfactant, and dissolving the mixture in a thermostatic chamber set at a temperature selected from a temperature range of 55 to 65 ° C to prepare a sol solution;

2) adding distilled water to the sol solution of step 1) and magnetically stirring to form a gel of a lyotropic liquid crystal;

3) gradually adding distilled water while continuing magnetic stirring to the gel of step 2) and applying shear force to disperse the gel into vesicle particles in water;

4) treating the suspended suspension of vesicle particles of step 3) with ultrasonic waves; And

5) cooling the suspension treated with the ultrasonic wave in step 4) to room temperature while stirring the magnet, and collecting the active ingredient, thereby producing a nanoflexible vesicle.

As used herein, the term "nano-flexible vesicle" refers to a nano-sized flexibly deformable vesicle, wherein the active ingredient is captured within the vesicle, And can act as a carrier to be absorbed.

A transparent sol solution in an isotropic state can be prepared by the above step 1). When the solution in the transparent sol state is left at room temperature, a thermotropic liquid crystal can be formed. When the step 1) is carried out at room temperature, a large amount of solvent is required to form a transparent sol state. However, at a high temperature (55 to 65 ° C), a transparent sol can be formed with only a very small amount of solvent. Unlike the case where a large amount of solvent is used, a sol formed by dissolving in an extremely small amount of solvent at a high temperature becomes a liquid crystal when it is cooled to room temperature. Although a sol of liquid crystal properties with a thermotropic liquid crystal can easily form a liquid crystal gel even when a small amount of distilled water is added at a high temperature, a sol having no heat and liquid crystal properties by adding a large amount of solvent can be used even when a large amount of distilled water is added It is difficult to make the desired vesicles in the next step because the liquid crystal gel is not well formed.

The active ingredient may be any peptide that can be used as a functional active ingredient of a cosmetic, and the functional active ingredient means a component exhibiting a functional effect in cosmetics such as skin whitening and wrinkle improvement.

In the above embodiments, the active ingredient may specifically be a peptide, more specifically an anti-aging peptide, more specifically a tetrapeptide, more specifically an anti-aging tetrapeptide, and more specifically, May be a tetrapeptide represented by Formula 1 or Formula 2, but is not limited thereto.

[Chemical Formula 1]

RR ¹ -Gly-Leu-Phe

(2)

R-Gly-Leu-Phe-R < ² >

(In Formula 1 and Formula 2, R is a tetra-decanoyl, and, Leu is D-Leu or L-Leu, Phe is D-Phe or L-Phe can be also R ¹ is of formula (I) and formula (2) Glycine, leucine, lysine, serine and R ² is selected from the group consisting of cystein, glutamic acid, phenylalanine, glycine, Wherein the amino acid is selected from the group consisting of isoleucine, valine, proline, leucine, serine, thrionine, tryptopane, tyrosine, The C-terminus may be in the form of a carboxylic acid or amide.

The tetrapeptide may be an anti-aging tetrapeptide consisting of an amino acid sequence of glutamine (Gln) -glycine-Leu-phenylalanine (Phe) and having a myristoyl acyl group at its terminal, But is not limited thereto.

Since the peptide used as the active ingredient is a substance having a high molecular weight formed by polymerization of at least three amino acids, percutaneous permeation absorption may be difficult.

The surfactant is preferably, but not limited to, sodium deoxycholate or polysorbate.

As used herein, the term "surfactant" may be a kind of elasticity-imparting component called an edge activator. Accordingly, the term " surfactant " and the term " edge activator " The surfactant can form a bilayer membrane of a vesicle while aligning molecules with the phospholipid. The surfactant has flexibility in the membrane by giving a free volume in which the phospholipid molecules can move in the array, Can be given. Since the surfactant has a relatively larger hydrophilic moiety than the lipophilic moiety and is advantageous in forming small spherical particles, when the surfactant is appropriately mixed with the phospholipid (for example, lecithin), the particle size of the vesicle .

The mixture of step 1) can be mixed at a weight ratio of active ingredient: phospholipid: surfactant = 1: 10: 98: 80: 1 to 10, more specifically, active ingredient: phospholipid: surfactant = To 80: 5 to 10, and may be mixed at a weight ratio of active ingredient: phospholipid: surfactant = 10: 85: 5, but is not limited thereto.

If the weight ratio of the active ingredient to the total weight of the mixture is decreased in the step 1), the active ingredient may be collected together with the self-assembling liquid crystal array of the phospholipid to form a vesicle membrane. However, if the weight ratio of the active ingredient is increased, The vesicle may not be formed properly because it acts as a foreign substance that interferes with the liquid crystal alignment of the phospholipid. Thus, it is preferred that the active ingredient does not exceed 10% by weight based on the weight of the phospholipid.

The solvent of step 1) may be any solvent selected from the group consisting of ethanol, propylene glycol, butylene glycol, and polyethylene glycol, preferably ethanol, but is not limited thereto. And any of known solvents used in the art in the manufacture of cosmetics can be used.

The concentration of the mixture to which the solvent is added in the step 1) may be 500 to 1500 mg / ml, specifically 600 to 1400 mg / ml, more specifically 700 to 1300 mg / ml, Specifically, it may be 800 to 1200 mg / ml, and may be, for example, 900 to 1100 mg / ml, but is not limited thereto. If it is out of the above range, a problem may arise that the liquid crystal is not well formed in the next step.

The thermostatic chamber of step 1) may be set to a specific temperature selected from a temperature range of 55 to 65 ° C and may be set to a specific temperature selected from a temperature range of 57 to 63 ° C, But it is not limited thereto. If the temperature is out of the above temperature range, the liquid crystal may not be well formed in the next step.

Milky white liquid crystal gel can be formed from the transparent sol solution by the above step 2).

The addition of the distilled water in the step 2) may be performed such that the volume ratio of the distilled water to the solvent added in the step 1) is 1: 0.5 to 1.5, for example, the volume ratio of ethanol: distilled water = 1: But is not limited thereto. If it is out of the above range, it may cause a problem that the vesicles are not dispersed well in the next step.

In the step 2), the magnetic stirring may be performed for 1 to 10 minutes, but the present invention is not limited thereto.

By the above step 3), the milky gel can be dispersed in water as vesicle-like particles to form a nano-sized flexible vesicle.

The addition of the distilled water in the step 3) may be carried out using a liquid transfer pump, but is not limited thereto.

Smaller and even vesicle particles can be formed by ultrasonication in step 4).

The ultrasonic treatment in the step 4) can be performed under the condition that the ultrasonic wave is applied for 1 minute by repeating the operation for 2 seconds and stopping for 1 second by using a probe type ultrasonic grinder (500 W).

By the above step 5), the nanoflexible vesicle membrane can be stabilized by transitioning from a rotational isomerization state to a highly ordered gel state.

The cooling of step 5) may be performed for 1 to 5 hours, but is not limited thereto.

The nano-flexible vesicles prepared according to one embodiment of the present invention can enhance percutaneous absorption of the collected active ingredients.

Nanoflexible vesicles prepared according to one embodiment of the present invention can be prepared by appropriately mixing an edge activator and ethanol with lecithin, which is a biocompatible material, and the produced vesicle particles have a very small size of several tens of nanometers And the vesicle membrane is flexible, which may be beneficial to passage through the horny layer of skin as compared to conventional vesicles.

In addition, the nano-flexible vesicle may be a transduction domain system (TDS) having an advantage of being easily collected and easily produced. In one embodiment of the present invention, an anti-aging peptide was collected on nano-flexible vesicles and skin permeation absorption experiments were carried out. As a result, the skin permeation absorption rate was about 4 times higher than that obtained when emulsified or collected in ordinary liposomes (FIG. 4) .

In general liposomes, a complicated process such as evaporation of a solvent or removal of a solvent by a high-pressure process is required. However, when the manufacturing method according to one embodiment of the present invention is used, the liquid crystal properties such as heat, The nanoflexible vesicles can be prepared by a relatively simple process including hydration and dispersion. In addition, when the above-described production method is applied to a hydrophilic active component, the collection efficiency can be significantly increased.

Further, another embodiment of the present invention is a cosmetic composition containing a nano-flexible vesicle which has collected active ingredients, more specifically, a nano-flexible vesicle obtained by capturing an active ingredient produced by a method according to one embodiment of the present invention Or a pharmaceutically acceptable salt thereof.

The kind of the additive which is further contained in the composition is not limited, and a common additive in the field of cosmetics may be added. Examples of common additives in the field of cosmetics include humectants, preservatives, thickeners, antibiotics, binders, disintegrants, diluents, lubricants, stabilizers, preservatives, fragrances, oils, water, surfactants, lower alcohols, chelating agents, For example, the composition may further comprise any one selected from the group consisting of moisturizers, preservatives, thickeners, and combinations of two or more thereof.

The humectant may be a conventional humectant used in the field of cosmetics, for example, glycerin or the like, but is not limited thereto.

The preservative may be a conventional preservative used in the field of cosmetics, for example, 1,2-hexanediol, caprylyl glycol / ethyl hexyl glycerin and the like. , But is not limited thereto.

The thickener may be a general thickening agent used in the field of cosmetics, for example, carbomer, simulgel, mixture of natrol / xanthan gum and the like, but is not limited thereto.

In one embodiment of the present invention, an enhancer, a preservative, and a moisturizer are added to a nano-flexible beacle in which a peptide is captured to produce essence cosmetics of various compositions. As a result, the more the amount of beacycell (i.e., (Table 3). The stability of the formulation was highest when simulgel EG was used, and the preservative type did not affect the stability of the formulation. The caprylyl glycol / ethyl hexyl glycerin mixed preservative used instead of parabens with controversial controversy showed almost equivalent flotation power to methylparaben against molds (Fig. 6).

The composition may be selected from the group consisting of softening lotion, astringent lotion, nutritional lotion, nutritional cream, massage cream, eye cream, eye essence, essence, cleansing cream, cleansing lotion, cleansing foam, cleansing water, pack, powder, body lotion, body cream, , A body cleanser, a hair dye, a shampoo, a rinse, a toothpaste, an oral cleanser, a regularizer, a hair preparation, a lotion, an ointment, a gel, a cream, a patch or a spray. However, It can have a formulation.

The cosmetic composition may cause a thixotropy phenomenon. In one embodiment of the present invention, the cosmetic formulation containing the nano-flexible beacle was found to have different shear stresses when increasing and decreasing the shear rate (FIG. 7). Therefore, the physical properties of the cosmetic formulation depend greatly on the rate and time of deformation, so that a feeling of use different from conventional cosmetic formulations can be obtained. That is, the cosmetic formulation which is not applied with the force is in the gel state, but when the cosmetic is applied to the skin by applying the force gradually, the formulation is changed to the sol state. When the application speed is increased, the viscosity of the formulation is decreased, . It also takes a while for the cosmetic formulation to be restored to the gel state after the application, so that the skin will feel soft and supple for a long time.

By using the nano-flexible vesicle production method of the present invention, it is possible to manufacture a vesicle having a size of several tens of nanometers having a flexible membrane, and thus the transdermal absorption of the active ingredient caught in vesicles can be enhanced. In addition, it is possible to manufacture vesicles by a simple process without a solvent evaporation process or a high pressure process, and the efficiency of collecting hydrophilic active ingredients can be significantly increased. The cosmetic formulation containing the nano-flexible beacle prepared using the above method exhibits a slight trophic property, and thus can be used for manufacturing a cosmetic composition having excellent feeling of use.

1 is a view illustrating a process for producing a nano-flexible beacle in which an anti-aging peptide is captured according to an embodiment of the present invention.
FIG. 2 is a graph showing changes in particle size (Vesicle Size, nm) of nanoflexible vesicles according to the mixing ratio of anti-aging peptide (A440) and lecithin (HPC).
3 is a Cryo-TEM photograph of a nanoflexible vesicle harboring an anti-aging peptide (A440).
FIG. 4 is a graph showing the skin penetration absorption enhancement effect of the anti-aging peptide (A440) by the nano-flexible vehicle according to an embodiment of the present invention; BG sol .: anti-aging peptide dissolved in butylene glycol solvent, EtOH sol .: anti-aging peptide dissolved in ethanol solvent, Emul: anti-aging peptide emulsified, Lipo: anti-aging Peptide, SDOC NFV: Anti-aging peptide captured in nanoflexible vesicles prepared using sodium deoxycholate (SDOC) as an edge activator, TW80 NFV: Nano-sized nano-particles prepared using polysorbate 80 as an edge activator Anti-aging peptide captured in a flexible vehicle.
FIG. 5 is a diagram illustrating a skin penetration absorption result of a cosmetic formulation containing a nano-flexible vial according to an embodiment of the present invention; FIG. Control (Emul.): Cosmetic formulations made with common emulsions, SDOC NFV: Cosmetic formulations containing nanoflexible vesicles made with sodium deoxycholate (SDOC) as an edge activator, TW80 NFV: Polysorbate with edge activator Cosmetic formulations containing nanoflexible vesicles prepared using a polysorbate 80.
FIG. 6 is a view showing a challenge test result of a black fungus of a cosmetic formulation containing a nano-flexible vise according to an embodiment of the present invention.
FIG. 7 is a graph comparing (a) cosmetic formulations containing a nano-flexible vial according to an embodiment of the present invention and (b) isopause phenomena of commercially available essence cosmetics.

Hereinafter, the present invention will be described in more detail in the following Examples. It should be noted, however, that the following examples are illustrative only and do not limit or limit the scope of the present invention. It will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

< Example  1> Preparation of materials

The anti-aging peptide is a myristoyl tetrapeptide (amino acid sequence: glutamine-glycine-leucine-phenylalanine) newly developed and synthesized by Miwon Company (manufacturer: Mi Won Company, trade name: A440_34; see Korean Patent Publication No. 1242488 (Hereinafter referred to as "A440") was used. Lecithin, which is the main component of Nano Flexible Vesicle, is Emulmetik950 (Lucas Meyer), which has a PC (Phosphatidyl Choline) component of 95% or more, by removing the unsaturated component by hydrogenating the lipid extracted from soybean. (Hydrogenated Phosphatidyl Choline). Sodium deoxycholate (Sigma Chem.), Polysorbate 80 (denoted TW80, Sigma Chem.), Sorbitan monooleate (denoted SP80, Sigma Chem.) And Cholesterol (denoted as CHOL) were used as the edge activator. Sigma Chem.) Were used. Ethanol used for dissolving lecithin and peptides was EP (Extra Pure; First Grade) grade. In addition, moisturizers, preservatives, thickeners and the like required for cosmetics were used for cosmetics.

< Example  2> anti-aging Peptides  Captured nano flexible Vesicle  Produce

1 g of a mixture of HPC, an anti-aging peptide, and an edge activator (anti-aging peptide and edge activator mixed up to 10 wt% each) and 1 ml of ethanol were placed in a round bottom flask, The inlet was closed with a stopper and completely dissolved in a thermostatic chamber at 60 ° C to form a transparent sol solution in an isotropic state. (A thermotropic liquid crystal is formed when a transparent sol solution is left at room temperature). In a transparent sol state, 1 ml of distilled water was added and the magnet was stirred for 2 minutes or more to form a milky gel in accordance with the lyotropic liquid crystal property. Next, in the dispersion step, distilled water was slowly added by using a constant amount pumping pump while applying a shear force while the agitator was continuously rotated to disperse the milky gel into vesicle-like particles in the water. This vesicle particle is a nano-flexible vesicle in which an anti-aging peptide is captured. Ultrasonic waves were applied to the suspension in which the vesicles were dispersed to make the vesicle particles even smaller and even with a size of several tens of nanometers. The supersonic suspension was slowly cooled at room temperature for about 2 hours under magnetic stirring to allow the vesicle membrane to stabilize in a highly ordered gel state in a rotational isomerization state. FIG. 1 summarizes the preparation process of a nano-flexible beacle containing an anti-aging peptide.

The general liposome needs a somewhat complicated process such as evaporation of solvent or high pressure process, whereas the nano-flexible vesicle according to the Embodiment 2 utilizes heat transfer, breast liquid crystal properties and liquid crystal phase transition appropriately, It is made up of a simple two-step process of dispersion. In addition, when such a process is applied to a hydrophilic substance, it has an advantage that the collection efficiency can be significantly increased.

< Experimental Example  1> Peptides Capture  Ratio Vesicle  Effect on particle size

The vesicle particle size according to the mixing ratio of peptide (A440) versus lecithin (HPC) was examined without adding an edge activator.

As shown in Fig. 2, the vesicle particle size increased as the ratio of A440 to HPC was increased. When A440 was mixed at a ratio of 10 wt% (at this time, the concentration of A440 was 2,000 ppm relative to the total solution), the vesicle particle size exceeded 200 nm. At the mixing ratio of more than 10 wt% of the peptide, the vesicle particles became a network gel rather than a dispersed state.

When the amount of the peptide is small, the peptide forms a vesicle membrane together with the self-assembled liquid crystal arrangement of the lecithin. However, when the amount of the peptide is more than 10 wt%, the peptide acts as a foreign substance that interferes with the liquid crystal arrangement of the lecithin The vesicles were not formed properly.

<Experimental Example 2> activator edge (Edge activator) is Effect on Vesicle Particle Size Experiments and Nanoflexible Determination of optimized composition of vesicle preparation

In order to make the particle size of the beacle smaller, different types of edge activators were added with different concentrations and the results shown in Table 1 were obtained. Table 1 shows changes in vesicle particle size according to the type and mixing ratio of the edge activator (E.A.). In particular, when SDOC and TW80 were added, the vesicles were greatly reduced to several tens of nano-sized particles, and SDOC was more effective than vesicle particles Which is effective in reducing the size of the image.

SDOC and TW80, used as edge activators, align molecules with lecithin to form bilayer membranes of vesicles, which give free volume to move lecithin molecules in the array Thereby imparting flexibility to the membrane. At the same time, the edge activator is advantageous to form small spherical particles because the hydrophilic head is relatively large (the critical packing parameter value is smaller) than the hydrophilic oil. Lecithin can reduce the size of vesicle particles by properly mixing the edge activator with lecithin, since it is difficult to make small spherical particles with a molecular structure close to one of the critical packing parameter values.

Considering that vesicle particles should be small while maximizing the concentration of A440, an optimized composition (based on a total of 50 g) in the manufacture of nano-flexible vesicles was determined as shown in Table 2 below. FIG. 3 shows a photograph of the finished vesicle particle confirmed by cryo-TEM.

< Experimental Example  3> Peptides  Captured nano flexible Vesicular  Skin penetration absorption experiment

In order to confirm the improvement of the skin permeation absorption when the anti-aging peptide (A440) was captured in the nano-flexible vesicles, the skin permeation absorption experiment using pig skin was performed as follows.

On the same day, the pork skin of the sirloin area, in which the subcutaneous fat was removed freely, was specifically ordered from the mangodong livestock wholesale market, cut into a size of 5 × 5 cm in area, and then the sample (A440 2,000 ppm concentration) 1 ml was evenly applied. After 5 hours at room temperature, the skin surface was rinsed with running water and the water was wiped dry. To remove the horny layer of skin, three times were repeated in such a manner that a strong scotch tape was pressed on the surface of the skin and then peeled off. The horny layer of the pig skin was cut into small pieces and then put into an ethanol-containing flask and magnetically stirred for a day to extract A440 which was absorbed into the skin. The extracted ethanol was filtered and injected into HPLC column to quantify A440 amount.

The results of the Nano-Flexible Vesicle (NFV) dramatically increasing the skin permeation absorption of A440 are shown in FIG. When A440 was emulsified with a common formulation (Emul.), A440 was dissolved in a solvent such as butylene glycol (BG sol), ethanol (EtOH sol) or the like which had the lowest penetration of skin permeation and was used as a skin penetration enhancer, (Lipo.), The absorption of A440 was about twice that of emulsion. However, when absorbed into the nano-flexible beacle, the skin permeation rate of A440 increased up to 4 times higher than that of solvent or liposome. In addition, when compared with the amount of A440 applied to the skin, the ratio of absorbed to the skin is only about 5% when dissolved in liposomes or solvents. However, when using Nano Flexible Vesicle (SDOC NFV or TW80 NFV) Absorption rate. SDOC was superior to TW80 when used as an edge activator.

In order for some foreign material to pass through the skin's horny layer and be absorbed deep into the skin, it must pass through the lipid layer between the keratinocytes, which is known to be several tens of nanometers in size. Generally, a peptide material having a high molecular weight is difficult to pass through the stratum corneum because the hydrodynamic volume of the peptide material when dissolved or emulsified in a solvent is larger than that of the lipid layer. Therefore, it is necessary to use a transdermal delivery system (TDS) for skin permeation of peptides. The nano-flexible beacle developed by the present inventor has tensile nano-size similar to that of the geologic layer and has good flexibility of the vesicle membrane and also has a good affinity with the skin lipid layer because the main component is phospholipid, thereby greatly increasing the skin permeation absorption of the peptide Is predicted.

< Experimental Example  4> Peptides  Captured nano flexible Vesicular  Manufacture of cosmetic formulations

Since nano-flexible beacles form a liquid crystal film together with components such as lecithin, edge activator, peptide, and ethanol by a self-assembly process, The vesicles may be destroyed or precipitation may occur. Therefore, it was attempted to develop a cosmetic formulation which can be stably preserved without phase separation. In order to minimize the influence of other possible mixing substances, only the thickener, moisturizing agent and preservative necessary for the cosmetic formulation were added to the vesicle suspension.

Specifically, an enhancer, a moisturizer, an antiseptic, and the like were added to the vesicle suspension to prepare a cosmetic formulation. Three types of increasing agents were used: carbopol 940, simulgel EG (Seppic, manufacturer), natrosol / xantan gum mixture. Carbomer, natrosol and xanthan gum were prepared by dissolving the solid powder in water to make a 2 wt% aqueous solution. Simulgel (EG) was used as the liquid product. Preservatives include parabens and phenoxyethanol, which are controversial in recent years, and they contain moisturizing and preservative effects such as 1,2-hexanediol, caprylyl glycol / ethyl hexyl glycerin, etc. And glycerin was used as a moisturizer.

The essence was prepared by the following procedure as a cosmetic formulation without oil component. First, the base formulation was prepared by dissolving the components other than vesicles, such as the thickening agent, preservative, moisturizer, etc., in water to prepare a base formulation. Nano-flexible vesicle solution (peptide concentration: 2,000 ppm) Followed by stirring to complete the formulation.

Tables 3 to 7 below show the composition of the essence formulation and the formulation stability results at room temperature and high temperature (45 DEG C). Tables 4 to 7 follow Table 3 and describe the same items as the left column of Table 3.

As a result, the stability of the essence formulation was lower as the content of the nano-flexible vesicle (peak concentration of A440) in which the peptide (A440) was collected was higher. At 0.1% concentration, phase separation occurred in most formulations except for very few formulations made with simulgel EG, whereas in the case of 0.001% -0.05% concentration, natrosol / xanthan gum xantan gum), and most of the other formulations were stable. It was found that the type of thickening agent greatly affected the stability of the formulation (simulgel EG> carbopol940> natrosol / xantan gum), but the kind of preservative did not significantly affect the stability of the formulation.

< Experimental Example  5> Peptides  Captured nano flexible Vesicle  Experiments on skin permeation absorption of cosmetic formulations

When the nano-flexible bicycloidal cosmetic formulation containing the peptide was made, the skin permeation absorption effect of the peptide was confirmed again.

First, cosmetic formulations made by semi-emulsification were prepared by mixing for 3 minutes with a homomixer while adding aqueous phase to the oil phase. The compositions of water phase and oil phase are as follows (Table 8).

Awards Paid ingredient Content (weight) ingredient content( weight% ) glycerin 18 ethanol 18.35 Xanthan gum 2% aqueous solution 7 Tween 80 1.5 Purified water 55 A440 0.15

In cosmetics containing Nano Flexible Vesicles, 75 g of a nano-flexible vesicle solution (A440 is a concentration of 2,000 ppm relative to the total solution) in which A440 was collected was mixed with 7 g of a 2% aqueous solution of xanthan gum 2% in water and 18 g of glycerin as a moisturizer Made into a formulation. In cosmetics made of ordinary emulsions and cosmetics containing nanoflexible vesicles, the concentration of A440 is 0.15%.

As a result of the experiment using pig skin in the same manner as in Experimental Example 3, cosmetic formulations containing nanoflexible vesicles (SDOC NFV or TW80 NFV) as compared with cosmetic formulations (Control (Emul.)) (Fig. 5). Fig.

< Experimental Example  6> Peptides  Captured nano flexible Vesicle  Cosmetic formulations Buoyancy  compare

Some of the cosmetic formulations containing the nano-flexible vesicles harvested from the peptides were selected and challenged against black fungus.

Specifically, samples of the cosmetic formulations used in Examples 4, 22, 25, 28 and 31 prepared in Experimental Example 4 were used. The challenge test was conducted as follows. 39 g of PDA (Potato Dextrose Agar) and 1 L of distilled water are added to an Erlenmeyer flask and dissolve well by applying heat. Sterilize in an autoclave at 121 ° C and 1.5 atm for 20 minutes. Cool the sterilized media to about 60 ° C, pour in about 40 mL of sterilized Petri dish in a clean bench, and harden to a solid plate medium. On the other hand, add 300 μl of fungicidal solution (5 × 10 ⁸ CFU) diluted per 10 g of cosmetics to 50 ml of falcon tube, mix well, and store in an incubator at 35 ° C. 100 μl of the cosmetics stored at intervals of 0, 1, 2, and 4 days are dispersed in an e-tube and dispersed with 10 times of distilled water, and 100 μl of the cosmetics is applied to a sterilized solid plate culture medium. The cultured medium was incubated for 48 hours in an incubator at 35 &lt; 0 &gt; C to observe the sterilization of the fungus.

As a result, in cosmetic formulations (Samples 22, 25 and 31) in which capryllyl glycol (0.2%) and ethylhexyl glycerin (0.2%) were mixed, , And a formulation with 0.2% of methylparaben (MP) prescribed (Sample No. 19 formulation added 0.04 g of methylparaben instead of 0.4 g of 1, 2-hexanediol). In the remaining formulations, Showing buoyancy that kills all fungi (Figure 6).

< Experimental Example  7> Peptides  Captured nano flexible Vesicle  Cosmetic formulations A trophy (Thixotropy) phenomenon

The thixotropy phenomenon has a different shear stress when the shear rate is increased and decreased. As shown in the figure, the area due to the difference in shear force occurs. As the area increases, the physical properties of the formulation This can greatly depend on the strain rate and time and can give a different feeling to the formulation.

The shear rate was increased to 20 (1 / s) at room temperature, and then the shear force was measured. Changes were observed.

As shown in FIG. 7, no cosmetic trophic phenomenon occurred in the commercially available essence cosmetics (Amore, Iope Moistogen Essential Serum) (FIG. 7B), but some cosmetic formulations 52, 55 containing nano- , 67) caused a slight trophic phenomenon (Fig. 7A).

From these results, the cosmetic formulations containing the nanoflexible vesicles containing the peptides were found to be in a gel state when stored, so that the physical stability of the formulation was maintained for a long time. When the composition was applied to the skin, it turned into a sol state, It was confirmed that this is an excellent advantage.

Claims (12)

1) ethanol is added to a mixture of the active ingredient, phospholipid and sodium deoxycholate and dissolved in a constant temperature bath set at a temperature selected from 55 to 65 ° C to prepare a sol solution Producing;
2) adding distilled water to the sol solution of step 1) and magnetically stirring to form a gel of a lyotropic liquid crystal;
3) gradually adding distilled water while continuing magnetic stirring to the gel of step 2) and applying shear force to disperse the gel into vesicle particles in water;
4) treating the suspended suspension of vesicle particles of step 3) with ultrasonic waves; And
5) cooling the suspension treated with the ultrasonic wave in the step 4) to room temperature while stirring the magnet, and collecting the active ingredient, thereby preparing a nanoflexible vesicle having a size of 20 to 50 nm,
Wherein the mixture of step 1) is mixed at a weight ratio of active ingredient: phospholipid: sodium deoxycholate = 1: 10: 98-80: 1 - 10. 2. The method of claim 1, wherein the active ingredient is a peptide used as a functional active ingredient of a cosmetic. 3. The method of claim 2, wherein the peptide is an anti-aging peptide. The method of claim 1, wherein the phospholipid is hydrogenated lecithin, unsaturated lecithin or lysolecithin. delete delete The method according to claim 1, wherein the concentration of the solvent to which the solvent is added in step 1) is 500 to 1500 mg / ml. 2. The method of claim 1, wherein the thermostatic chamber of step 1) is set to a temperature of 60 &lt; 0 &gt; C. The method of claim 1, wherein the nano-flexible beacle enhances transdermal absorption of the captured active ingredient. 9. A cosmetic composition comprising a nanoflexible vesicle harvested by the method of any one of claims 1 to 4 and 7 to 9, [Claim 11] The cosmetic composition according to claim 10, wherein the composition further comprises any one selected from the group consisting of moisturizers, preservatives, thickeners, and combinations of two or more thereof. 11. The composition of claim 10, wherein the composition is selected from the group consisting of softening agents, convergent lotion, nutritional lotion, nutritional cream, massage cream, eye cream, eye essence, essence, cleansing cream, cleansing lotion, cleansing foam, cleansing water, Wherein the composition is a formulation of a body cream, a body essence, a body cleanser, a hair dye, a shampoo, a rinse, a toothpaste, an oral cleanser, a regularizer, a hair dye, a lotion, an ointment, a gel, a cream, a patch or a spray.

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