KR100479010B1 - Methods of hair removal using ALA with cationic liposome - Google Patents

Abstract

The present invention relates to an external preparation for hair removal prepared by mixing 5-aminolevulinic acid and a cationic liposome, and a method for hair removal using the same. In the present invention, 5-aminolevulinic acid, which is an acidic cationic drug, is a simple liposome. By formulating with non-cationic liposomes, it is possible to reduce its own toxicity without altering the ionicity of the drug and to increase the drug uptake of the cells. In addition, since the liposomes have a cation, iontophoresis can be applied, and thus, a simple application of nonionic or neutral liposomes could provide an unexpected effect.

Description

External preparation for hair removal comprising ALA and cationic liposomes {Methods of hair removal using ALA with cationic liposome}

The present invention relates to an external preparation for hair removal prepared by mixing 5-aminolevulinic acid (hereinafter abbreviated as "ALA") and cationic liposomes, and a method for hair removal using the same. More specifically, by formulating an acidic cationic drug ALA into a cationic liposome rather than a simple liposome, it can reduce its own toxicity without changing the ionicity of the drug and increase the drug absorption of cells. The present invention relates to an external preparation and a method for depilation using iontophoresis using the preparation, and a method of applying heat and light together.

Removing unwanted hair is called epilation, which includes temporary epilation and permanent epilation. Temporary hair removal methods include shaving, plucking, applying epilation creams, and applying tapes on and off. On the other hand, permanent hair removal is a procedure that selectively destroys only hair follicle cells that make hair and permanently removes hair without damaging the skin. There are electric hair removal and laser hair removal.

Electroepilation is a method of inserting a thin needle into the pores to reach the hair follicles, the root of the hair, and then destroying the hair follicles by passing a weak current. However, this method has to find the pores one by one, so it takes a lot of time, and the current flow is long, it is easy to leave scars and anesthesia has to be disadvantaged.

Laser hair removal is a method of damaging hair follicles by focusing laser heat on the hair roots and the melanin around them by using the laser's response to melanin. However, if the skin color is exceptionally black or recently tanned, the amount of melanin pigment in the epidermis increases, so as the amount of laser light is absorbed into the epidermis increases, hair does not reach a sufficient amount and the effect of laser treatment Will be reduced. In addition, the hair of the body has a period of growth, degenerative phase, rest period, laser hair removal will be effective only on the growth phase hair. Usually hair growth rate is about 30% in the body, so the hair removal is not completed with one laser treatment, but it is necessary to wait 3-5 times to wait for the current degenerative or resting hair to change into hair growth hair.

On the other hand, photodynamics treatment using ALA has been used for skin diseases such as actinic keratoses, psoriasis, warts (verrucae) and skin cancer. ALA is applied to the skin to be absorbed by epithelial cells to produce protoporphyrin IX (hereinafter abbreviated as " PpIX "), a heme precursor. PpIX is a very effective photoresist that is excited in the triplet state by light energy and reacts with surrounding oxygen to produce generator oxygen. The generated generator oxygen can cause damage to cell membranes and cell death, thereby treating skin diseases such as the above.

Photodynamic therapy with ALA is also being applied to permanent hair removal. That is, when hair is removed or removed with a depilatory agent and ALA is applied to the skin, ALA penetrates into the hair follicles through the pores and the hair follicle cells synthesize PpIX to show red fluorescence (see FIG. 1). When light at 600-700 nm wavelength is irradiated, the hair follicle cells are damaged by excitation of PpIX . The problem is whether it is possible to selectively transport large amounts of ALA into the hair follicles, and that light of 600-700 nm wavelength is difficult to penetrate to the depth of the hair follicles.

On the other hand, liposomes point to microvesicles having a hydrophilic space inside and a closed double lipid membrane outside. Liposomes contain water-soluble molecules or drugs in the central hydrophilic space, and lipid-lipid membranes can attach fat-soluble drugs or combine positive and negative charges. The biggest characteristics of liposomes are diversity and elasticity. In other words, liposomes are easy to change the structure, by providing a completely new physical properties, functions, uses by different structures and components. Herein, liposomes have the advantages of biocompatibility, degradability, and stability. Due to these advantages, liposomes are widely used in numerous substances and drug delivery materials. Substances that can be delivered by binding to liposomes include peptides, proteins, fats, such as anticancer drugs (adriamycin), antifungal drugs (amphotericin B), antibacterial agents, immunomodulators, antigens and antibodies, contrast medium, hemoglobin, growth factors, etc. And DNA, etc., is applied in most clinical practice.

An object of the present invention is an external preparation for hair removal agent which can simultaneously solve problems such as selectivity, penetration depth and penetration time for the application site of the drug while reducing its own toxicity without changing the ionicity of the ALA drug for hair removal agent. It is to provide a depilation method used.

In order to achieve the above object, in the present invention, an external preparation for hair removal prepared by mixing an acidic ALA and a cationic liposome, and a method for applying hair removal using ionic osmotherapy using the preparation and applying heat and light together to provide.

Hereinafter, the present invention will be described in detail.

In the present invention, first, a 33-67% by weight of ALA and 33-67% by weight of cationic liposomes are mixed to provide an external preparation for hair removal.

In the preparation of the external preparation, it is added at a concentration of 5 to 20% by weight of ALA and 5 to 20% by weight of the cationic liposome with respect to the aqueous solution used as the solvent, and as a result, an external preparation for hair removal having the composition described above is provided.

The cationic liposome is preferably prepared by mixing a lipid and a cationic additive constituting the neutral liposome.

At this time, the lipid constituting the neutral liposome can be used such as DG, PC, POE or CH, cationic additives can be used such as DAP, TAP, DOS, SM or PS, these are specifically shown in Table 1 below .

TABLE 1

Compound name Acyl Group's Carbon Chain Molecular Weight Geology DG 1,2-diacyl - sn - glycerol (1,2-Diacyl- sn -glycerols) 10: 0, 12: 0, 14: 0, 16: 0, 18: 1 400.6-620.99 PC 1,2-diacyl-sn-glycero--phosphocholine (1,2-Diacyl- sn -glycero-phosphocholine ) 10: 0, 12: 0, 14: 0, 16: 0, 18: 1 565.7 ~ 786.2 POE Polyoxyethylene (10) acyl ether 16: 0, 18: 0, 18: 1 682.9 to 711.0 CH Cholesterol 386.66 Cationic additive DAP 1,2-Diacyl-3-dimethylammonium propane 14: 0, 16: 0, 18: 1 539.9 ~ 648.1 TAP 1,2-Diacyl-3-trimethylammonium propane 14: 0, 16: 0, 18: 1 590.4 to 698.6 DOS 1,3-Diacyl-2-carboxyspermyl-propylamide 18: 1 1,089.4 SM Spermine 348.2 PS Protamine sulfate 5,000 to 10,000

The preparation is preferably prepared by mixing ALA, lipids and additives from the beginning when the cationic additive is a cationic lipid such as DAP, TAP or DOS, and when the cationic additive is a water-soluble additive such as SM or PS, After preparing the formulation of the lipid, it is preferred to add further additives to complete the formulation.

Cationic liposomes are preferably 80 to 90% by weight: 10 to 20% by weight of the mixing ratio of the lipid and the cationic additive constituting the neutral liposome.

Preferred examples of the cationic liposomes are prepared by mixing DG / CH / POE / cationic additives in a ratio of 50 to 60/15 to 20/20 to 30/10 to 20, or adding PC / CH / cationic additives to 60 by weight. It manufactures by mixing in a ratio of -70/20-30/10-20.

The present invention can be used by directly applying the formulation formulated with only ALA and cationic liposomes as described above directly to the area to be epilated, a natural oil (oil), fatty acids (fatty acid), etc. as a carrier suitable for transdermal administration, if necessary Formulations may be further prepared to prepare external preparations. The external preparations of the present invention may be prepared by various methods, for example ointments, transdermal patches, gels, creams, lotions, solutions, powders ( powders, cataplasma or pastes, and the like.

In addition, the present invention provides a hair removal method using the external preparation of the present invention, the method comprising the step of removing the hair of the area to be depilated, the above-described external preparation for hair removal of the present invention to the site where the hair is removed Applying.

In addition to the above process, the hair removal method of the present invention comprises the steps of: conducting iontophoresis at the site of application of the external preparation for the hair removal agent; And simultaneously radiating heat and light to the site. At this time, the light is a light ray having a wavelength of 600 ~ 700 nm, the heat is preferably infrared.

The iontophoresis method may be applied for 10 to 20 minutes at a current of 0.1 to 1.0 mA / cm 2 by connecting a (+) pole to the site where the external preparation for hair removal is applied and a (-) pole to the opposite side of the site. .

Since hair follicles need to be destroyed for permanent hair loss, in the present invention, ALA was prepared as a cationic liposome and transported by iontophoresis to improve its selectivity and efficiency.

ALA, an epileptic drug, is an acidic cationic drug. Therefore, the preparation of a cationic liposome rather than a simple liposome can reduce its own toxicity without changing the ionicity of ALA.

In addition, since human skin generally has a weak acidic pH, cationic liposomes are more permeable or absorbent than nonionic or neutral liposomes. Moreover, ALA is an acidic cationic substance as described above, and the activity is greatly reduced when the pH of the solution is neutral or acidic. In view of this, cationic liposomes not only serve as carriers, but also have the advantage of preserving and increasing the function of ALA. Thus, PpIX expression in hair follicles was significantly increased thanks to the function of ALA, which was further enhanced.

ALA formulated with cationic liposomes were excellent in permeability and increased ALA uptake of cells. Furthermore, in the present invention, the ionosmotherapy can be applied by using the property that the liposome has a cation. As a result, ALA was rapidly delivered to the hair follicles deep into the skin as well as sebaceous glands, and the expression of PpIX was evident, which was not found in the simple application of nonionic or neutral liposomes. In view of this, cationic liposomes maintain the ionicity of ALA, which increases the function of ALA. By applying iontophoresis, existing problems related to selectivity, depth of permeation, and time of permeation can be solved simultaneously. .

In addition, in the present invention, after ALA was sufficiently absorbed and PpIX was expressed, it was possible to damage hair follicles by using both natural light and infrared light. Conventional photodynamic therapy of hair removal mainly uses a light source and a laser that emits light, but in the present invention, the hair removal effect can be further enhanced by using light and heat simultaneously. Furthermore, the application of heat at the same time was more effective than the application of light alone, with the additional effect that the heat in the red zone was anti-inflammatory. That is, the method of the present invention which removes the hair of the desired area for hair removal, and then applies 5-20% ALA formulated with cationic liposomes, applies iontophoresis and simultaneously exposes the hair follicles deep into the skin, By effectively destroying the skin side effects can be minimized, and the effect of hair removal treatment can be greatly improved.

Hereinafter, the present invention will be described in detail by examples. However, the examples are only to illustrate the invention and the present invention is not limited by the following examples.

Liposomes and cationic additives used in the examples are shown in Table 1 above.

<Preparation Example> Manufacturing method of external preparation for hair removal agent of this invention

Liposomes were first mixed with lipids and dissolved at 70 ^° C. according to the neutral liposome preparation composition. The melted solution was placed in a preheated syringe and connected to an injection set to which a syringe containing an aqueous solution such as saline containing ALA or 5% dextrose was connected. Subsequently, both solutions were moved back and forth several times using a syringe, and the mixture was sufficiently mixed with lipids, followed by sonication for 3-5 minutes at room temperature. The concentration of liposomes in the aqueous solution is 50-200 mg / ml, ALA is 5 to 20% by weight based on the weight of a certain aqueous solution. Cationic liposomes are formulated as ALA by adding them together with lipids from the beginning when the additive is cationic lipids such as DAP, TAP or DOS, and mixing the lipid-ALA first when water-soluble additives such as SM or PS, Formulated by addition.

The external preparations for the depilatory hair in various composition ratios were prepared by the preparation method as described above.

Example 1 Preparation of Cationic Liposome-ALA (20%)

DG (12: 0)... … … … … … … … … … … … … … 50 mg

CH… … … … … … … … … … … … … … 15 mg

POE… … … … … … … … … … … … … … 20 mg

Cationic additive (TAP). … … … … … … … … … 15 mg

The external preparation of the present invention was prepared by the method described in the above formulation example in 20% by weight of ALA (200 mg) and 10% by weight of the cationic liposome of the above composition in 1 ml of saline.

Example 2. Preparation of Cationic Liposome-ALA (20%)

PC (14: 0)... … … … … … … … … … … … 140 mg

CH… … … … … … … … … … … … 40 mg

Cationic additive (SM). … … … … … … … … 20 mg

The external preparation of the present invention was prepared by the method described in the formulation example in 20% by weight of ALA (200 mg) and 20% by weight of the cationic liposome of the above composition in 1 ml of 5% dextrose.

Example 3. Preparation of Cationic Liposome-ALA (15%)

DG (14: 0)... … … … … … … … … … … … … … 75 mg

CH… … … … … … … … … … … … … … 25 mg

POE… … … … … … … … … … … … … … 30 mg

Cationic additive (PS). … … … … … … … … … … 20 mg

The external preparation of the present invention was prepared by the method described in the formulation example in 15% by weight of ALA (150 mg) and 15% by weight of the cationic liposome of the above composition in 1 ml of saline.

Example 4. Preparation of Cationic Liposome-ALA (10%)

DG (12: 0)... … … … … … … … … … … … … … 50 mg

CH… … … … … … … … … … … … … … 15 mg

POE… … … … … … … … … … … … … … 20 mg

Cationic additive (TAP). … … … … … … … … … 15 mg

The external formulation of the present invention was prepared by the method described in the formulation example in 10% by weight of ALA (100 mg) and 10% by weight of the cationic liposome of the above composition in 1 ml of saline.

Example 5. Preparation of Cationic Liposome-ALA (5%)

DG (14: 0)... … … … … … … … … … … … … … 25 mg

CH… … … … … … … … … … … … … … 8 mg

POE… … … … … … … … … … … … … … 10 mg

Cationic additive (DAP). … … … … … … … … … 7 mg

5 wt% ALA (50 mg) and 5 wt% of the cationic liposomes of the above composition were prepared in 1 ml of saline, and the external formulation of the present invention was prepared by the method described in the formulation example.

Experimental Example 1. Entrapment efficiency (EE) and stability

The capture efficiency of ALA surrounded by liposomes decreased (EE percent) at lower lipid concentrations, but was about 40-60% at baseline concentrations of 100 mg / ml. In the case of liposome-drug used for skin application, unlike the intravenous injection, the separation of only the enclosed drug is not urgently needed, so the whole liposome solution after the preparation was used for the treatment. After the preparation of the prepared cationic liposome-ALA preparation, the result was stored in the refrigerator for about one month, and both liposomes and ALA drugs were stable and generally stable.

Experimental Example 2. Animal Experiment

(1) skin application

The results after the application of various ALA to the hair of a rat was shown in FIG. 2. Little red fluorescence was detected in the control group to which ALA was not applied (see FIG. 2A). In the simple application of ALA, PpIX expression was observed in sebaceous glands after 2 hours, peaked after 8 hours, and almost no red fluorescence was observed after about 24 hours. Comparing the application of ALA alone without liposomes (see FIG. 2B) and the application of ALA prepared with cationic liposomes (see FIG. 2C), the red fluorescence of PpIX was detected about three times higher. As mentioned earlier, this effect is due to the excellent permeation and absorption of cationic liposomes into the skin and the enhancement of ALA function due to increased cationicity.

(2) iontophoresis

In a shorter time, a large amount of ALA was delivered to the hair follicles of the dermis and iontophoresis was applied for high PpIX expression. The instrument used was Ionzyme (Environ Cosmeceutics International, Japan), and the intensity of the current used was 0.1-1.0 mA / cm 2. The application side was applied with the positive pole and the other side with ultrasonic gel and the negative pole was connected. After about one hour of preliminary coating, ion osmosis was performed. Since it was already formulated with cationic liposomes in consideration of the ionicity of ALA, the positive electrode was connected to the applicator and the negative electrode was connected to the opposite side. In rats, the most suitable delivery was achieved by application for 15-20 minutes at 0.25 mm 3 / cm 2. The ALA drug itself has a cation, so iontophoresis is possible (see FIG. 2D), but the cationic liposome-ALA (see FIGS . Significantly lower in the amount of expression. The use of water soluble cationic additives (SM and PS) (see FIGS. 2F and 2G), rather than the use of cationic lipid additives (TAP) (see FIG. 2E), was similar. Some standardization was required for each cationic liposome preparation, but both were satisfactory in terms of time and efficiency.

(3) hair follicle necrosis by heat and light

ALA (Example 1) prepared with cationic liposomes on the back of the rat was removed, coated with gauze having a width and length of 1.5 cm, and then subjected to iontophoresis. Ultrasonic gel was applied to the abdomen where the hair had already been removed, and the negative electrode was connected and the positive electrode was connected to the cationic liposome-ALA drug, followed by ion osmotherapy for 15 minutes at a current intensity of 0.25 V / cm 2. After 2 hours, the infrared rays were exposed for 30 minutes in the state exposed to natural light, and the result was observed for 4 weeks. Compared to the control group (see FIG. 3a), the cationic liposome-ALA comparison group (see FIG. 3b) showed depilation effect from 2 weeks, and at 3 weeks, only the depilation effect of the liposome-drug site was clearly seen. there was.

Changes in hair follicles after 1 week of exposure to natural light and infrared light for 10 minutes (see FIG. 4B) or 30 minutes (see FIG. 4C) after cationic liposome-ALA application / ion osmotherapy were examined. Necrosis of hair follicles could be observed in the ALA and heat / light applied areas compared to the control group (see FIG. 4A).

As described above, using an external preparation for a hair removal agent comprising the ALA and the cationic liposome of the present invention increases the selectivity to the site of application of the drug while reducing its own toxicity without changing the ionicity of the ALA, Large amounts of ALA could be selectively delivered to the hair follicles, such as being able to penetrate the drug to the desired depth during the infiltration time.

1 is a schematic diagram of pore penetration of ALA.

Figure 2a is a photograph (control group) showing the pore state when nothing is applied to the skin.

Figure 2b is a photograph showing the pore state when only ALA is applied to the skin alone.

FIG. 2C shows the use of an external preparation of the present invention (neutral liposome (hereinafter abbreviated as “N”) and cationic additive (TAP) as the cationic liposome on the skin); Hereinafter, denoted as "N / TAP"] is a photo showing the pore state when the coating.

Figure 2d is a photograph showing the pore state when only ALA is applied to the skin alone and then iontophoresis is performed.

Figure 2e is a photograph showing the pore state when the application of the external preparation (N / TAP) of the present invention to the skin and then subjected to iontophoresis.

Figure 2f is a photograph showing the pore state when the application of the external preparation (N / SM) of the present invention to the skin followed by iontophoresis.

Figure 2g is a photograph showing the pore state when the application of the external preparation (N / PS) of the present invention to the skin and then subjected to iontophoresis.

Figure 3a is a photograph showing that the hair grows again over time when no treatment after removing the hair of the rat.

Figure 3b is a photo showing that the hair grows over time when treated with the manufacturing method of the present invention after removing the hair of the rat.

4A to 4C are photographs showing the degree of destruction of hair follicles due to activation of ALA when treated with a control group and the method of the present invention.

Claims (14)

33-67 wt% of 5-aminolevulinic acid and 1,2-diacyl- sn -glycerol (DG), 1,2-diacyl- sn -glycero-phosphocholine (PC) , 1,2-diacyl-3-dimethylammonium propane (DAP), 1,2-diacyl-3-trimethyl and a lipid selected from the group consisting of polyoxyethylene (10) acyl ether (POE) and cholesterol (CH) Cationic liposomes prepared by mixing cationic additives selected from the group consisting of ammonium propane (TAP), 1,3-diacyl-2-carboxycisyl-propylamide (DOS), spermine (SM) and protamine sulfate (PS) The external preparation for hair removal which is formulated by mixing 33-67 weight%. delete delete delete The method of claim 1, The formulation is an external preparation for hair removal, characterized in that the additive is prepared from the beginning and mixed with 5-aminolevulinic acid and lipid when the cationic lipid. The method of claim 1, When the additive is a water-soluble cationic additive, after the preparation of 5-aminolevulinic acid and a lipid preparation, further comprising a water-soluble cationic additive to complete the formulation for hair removal. The method of claim 1, The mixing ratio of the lipid and the cationic additive is 80 to 90% by weight: 10 to 20% by weight of the external preparation for hair removal. The method of claim 1, The cationic liposomes comprise 1,2-diacyl- sn -glycerol (DG) / cholesterol (CH) / polyoxyethylene (10) acyl ether (POE) / cationic additive in a weight ratio of 50 to 60/15 to 20/20 to A hair removal preparation for external use, which is prepared by mixing at a ratio of 30/10 to 20. The method of claim 1, The cationic liposome is prepared by mixing 1,2-diacyl- sn -glycero-phosphocholine (PC) / cholesterol (CH) / cationic additive in a weight ratio of 60 to 70/20 to 30/10 to 20. External preparation for hair removal, characterized in that. delete delete delete delete delete