KR20160001715A - Liposome included horse oil, the method preparing thereof and composition for treating burn or uv-cut containing the same - Google Patents

Abstract

The present invention relates to a process for the preparation of a pharmaceutical composition comprising: i) melting a marrow at 40 to 45 캜; ii) mixing the melted horse meat oil with lecithin and an emulsifying aid to dissolve in purified water; And iii) injecting the dissolution mixture of step ii) into a high-pressure emulsifier and emulsifying the emulsion three to five times at a pressure of from 15,000 to 30,000 psi to obtain a liposome encapsulated with the emulsion; And a method for producing a liposome of the present invention. The present invention also relates to a hair cosmetic liposome and a cosmetic composition for treating or UV-screening comprising the liposome as an active ingredient. The present invention also relates to a hair oil-containing liposome and a pharmaceutical composition for treating burns or ultraviolet rays comprising the same as an active ingredient.

Description

TECHNICAL FIELD [0001] The present invention relates to a liposome-containing liposome, a method for producing the liposome, and a method for preparing the liposome, which comprises the liposome-encapsulated liposome,

The present invention relates to a liposome encapsulated with a liposome, a method for producing the liposome, and a liposome encapsulated with a liposome as an active ingredient.

Mayu is an oil of the horse, and it is said that it is rich in vitamin E from the past and it is beneficial for the prevention of fine lines and aging, and it has been used for skin beauty for a long time in folk remedies. For example, it has been known that hemp is very effective for burning, which is effective for burning and relieving inflammation. It contains a large amount of palmitoleic acid, which has strong antimicrobial action, as a major component of sebum to protect human skin It is known to form a natural protective film of the skin to give the skin a high moisturizing effect. It also contains a large amount of Vitamin E, which makes the blood circulation vigorous, so that the oil itself can serve as a good cosmetic and is also used in skin cosmetics. In addition, Maou is the closest component to sebum, and because it has almost the same composition ratio of fatty acids, oil and moisture, it has high affinity with human skin and contains about 60% or more of unsaturated fatty acids, It is known as a substance.

Mayu is used as a treatment for atopic and burns by folk remedy, and it shows the improvement of the skin of Mayu in the pharmacology book of Chinese Taipei University. In addition, each household in Japan is using Mayu as a skin care home remedy. However, prior to the use of Mayu as a reliance on folk remedies, more scientific verification of the effectiveness of burn healing is needed.

Liposomes are emulsions of the W / O / W type proposed by Bangsu in the 1960s [J. Mol. Biol., 13, 238 (1965)), wherein amphiphilic phospholipids are self-aligned by hydrophobic forces in the aqueous phase. When liposomes are used as medicines or cosmetics, they have the advantages of (a) a biocompatible component of phospholipid, which can be decomposed in vivo and not toxic, (b) (C) the size and shape characteristics of the liposome can be controlled and adjusted according to the production method, surface charge, lipid composition, etc., and (d) (E) If the inclusion substance is a drug, it can be targeted to a specific tissue, thereby enhancing the therapeutic effect of the drug and administering the drug through almost any route of administration (oral, subcutaneous, intravenous, intramuscular or intraperitoneal injection) There are many advantages of.

Korean Patent Laid-Open Publication No. 2010-0000922 discloses a composition for treating or preventing atopic dermatitis, which contains refined margarine as an active ingredient. However, it is difficult to penetrate the skin with only refined margarine, which is a problem in its efficiency.

Accordingly, the present inventors have scientifically verified the efficacy of Mayu by developing a liposome of Maou, which is easy to penetrate into the skin.

Korea Patent Publication No. 2010-0000922

An object of the present invention is to provide a liposome-containing liposome.

Another object of the present invention is to provide a pharmaceutical composition for treating burns or ultraviolet ray shielding comprising a maleic in-plant liposome as an active ingredient.

It is still another object of the present invention to provide a cosmetic composition containing a maleic graft liposome as an active ingredient.

It is still another object of the present invention to provide a method for producing a horse oil inclusion liposome.

In order to achieve the above object,

i) melting the marrow at 40 to 45 占 폚;

ii) mixing the melted horse meat oil with lecithin and an emulsifying aid to dissolve in purified water; And

iii) injecting the dissolution mixture of step ii) into a high-pressure emulsifier and emulsifying the emulsion three to five times at a pressure of 15,000 to 30,000 psi to obtain a liposome encapsulated with the emulsion;

The present invention provides a method for producing a liposome of the present invention.

As used herein, the term " mayonnaise " is a fat component extracted from the fatty tissue of horses and includes palmitoleic acid and ceramide as main ingredients of maruyu. Mayu has the effects of moisturizing, protecting skin, promoting cell regeneration, UV protection, and antibacterial activity. In the embodiment of the present invention, the Mayu can be Jeju Mayu extracted from Jeju Mountain Horse.

In an embodiment of the present invention, the lecithin functions to form a stabilized lipid membrane by stably encapsulating a horse oil, and in particular, it can be selected from among hydrogenated lecithin, unsaturated lecithin and lysolecithin.

In the embodiment of the present invention, the emulsifying aid contributes to the formation of the multilamellar structure as well as the stability of the liposome. Generally, a higher fatty alcohol may be used, and preferably cetyl alcohol, stearyl alcohol, Allyl alcohol and the like can be used or a fatty acid having 6 to 22 carbon atoms such as caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, arachidonic acid, Hexanoic acid) or a pharmaceutically acceptable salt thereof (e.g., sodium salt, potassium salt, calcium salt), albumin, textur, and the like.

In the embodiment of the present invention, the dissolution mixture in step ii) may be contained in an amount of 10 to 40% by weight of a hemp, 1.0 to 4.0% by weight of lecithin, 0.5 to 3.0% by weight of an emulsifying aid, and 55 to 85% 20 to 30% by weight of trehalose, and 2 to 3% of lecithin.

At this time, when the amount of the talc is out of the range of 10 to 40% by weight, the therapeutic treatment or ultraviolet ray blocking effect of the talc inoculum liposome may be weak. If the content of lecithin is less than 1.0% by weight, emulsification may not be performed. If the content of lecithin is more than 4.0% by weight, liposomes may change and liposomes of desired sizes may not be obtained. If the content of the emulsifying aid is less than 0.5% by weight, emulsification may not be performed. If the amount of the emulsifying aid is more than 3.0% by weight, the expected effect of the liposome may be insufficient.

In the embodiment of the present invention, after melting the horse oil at 40 to 45 ° C, the melted horse oil is firstly emulsified by mixing with lecithin and an emulsifying aid and dissolving in purified water. Thereafter, the dissolution mixture is injected into a high-pressure emulsifier and subjected to secondary emulsification, whereby a liposome encapsulated with the final emulsion can be prepared. At this time, the high-pressure emulsifier can be operated at a pressure of 15,000 to 30,000 psi, and the number of emulsions can be emulsified 3 to 5 times.

When the number of times of emulsification is 3 to 5 times, the grain size of the grafted liposome can be adjusted to 0.25 to 0.5 탆. In the case of producing a horse oil inclusion-forming liposome within the above-mentioned particle size range, the effect of the image treatment or ultraviolet ray shielding can be maximized.

In the embodiment of the present invention, when the content of the talc is set to 20 wt% and the number of emulsions is set to three, the grain size average may be about 0.43 mu m. When the content of the talc is set to 25 wt% and the number of emulsions is set to 4, the particle size average may be about 0.37 mu m. When the content of the talc is set to 30 wt% and the number of emulsions is set to 5, the particle size average may be about 0.31 mu m. Preferably, a uniform liposome particle of about 0.3 탆 can be obtained when the content of the present invention is adjusted to 20% by weight and the emulsification frequency is set to 5 times.

The present invention also relates to

The emulsion composition of the present invention is prepared by emulsifying a dissolution mixture containing 10 to 40% by weight of lecithin, 1.0 to 4.0% by weight of lecithin, 0.5 to 3.0% by weight of an emulsifying aid and 55 to 85% by weight of purified water with a high pressure emulsifier to a particle size of 0.25 to 0.5 μm By weight of the liposome.

The present invention also provides a cosmetic composition comprising the above male oil-in-blended liposome as an active ingredient. The cosmetic composition may be used for burn treatment or ultraviolet ray shielding.

The cosmetic composition of the present invention can be used in cosmetics such as softening lotion, convergent lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, paste, gel, cream, lotion, powder , Formulations selected from the group consisting of soaps, oils, foundations, waxes, sprays, body lotions, body creams, body oils and body essences.

The cosmetic composition may further contain, in addition to the liposome-containing liposome of the present invention, a lipid, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, , Water, ionic or nonionic emulsifiers, fillers, sequestering agents and chelating agents, preservatives, vitamins, barrier agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics And may contain adjuvants conventionally used in the cosmetics field, such as any other ingredients used.

The present invention also provides a pharmaceutical composition for the treatment of burns or ultraviolet rays, which comprises the above-mentioned horse oil inclusion liposome as an active ingredient.

The pharmaceutical composition according to the present invention can be used as an active ingredient in a mammalian inclusion liposome as it is or in the form of a pharmaceutically acceptable salt. The salt is not particularly limited as long as it is pharmaceutically acceptable so long as it is pharmaceutically acceptable and includes, for example, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid acetic acid, tartaric acid, lactic acid, citric acid, fumaric acid, , Benzenesulfonic acid, toluenesulfonic acid, and naphthalenesulfonic acid. In addition to acid addition salts, additional base salts such as sodium hydroxide, potassium hydroxide, triethylamine, tertiary-butylamine may also be used.

In the formulation of the pharmaceutical composition according to the present invention, the content of the active ingredient may vary widely depending on the formulations, and is according to a conventional method.

The pharmaceutical composition according to the present invention may be formulated by adding various auxiliaries used in medicine such as carriers and other additives such as stabilizers, emollients, emulsifiers and the like, if necessary, unless adversely affecting the active ingredients.

In addition, the pharmaceutical composition according to the present invention can be administered parenterally or orally. As the route of parenteral administration, transdermal administration is preferable, and local application is most preferable. For example, it may be manufactured in the form of a bandage, but is not limited thereto. The formulations may be any formulations suitable for pharmaceutical preparations including ointments, creams, injections, powders, granules, tablets, and the like.

The preferred dose of the pharmaceutical composition according to the present invention may be 0.001 to 1000 mg / Kg · day, but is not limited thereto. In addition, the composition according to the present invention may be administered alone, or may be administered together with other medicines in order to aid in the same or another medicament. In addition, in the composition of each formulation, components other than the composition, which is the above-mentioned essential ingredient, can be mixed and selected by a person skilled in the art according to the formulation or purpose of use of the other external preparation. In this case, Can happen.

In one embodiment of the present invention, the composition containing the horse oil-in-blended liposome as an active ingredient was treated with a mouse irradiated with ultraviolet rays to confirm the healing efficacy of horse oil. In addition, in the embodiment of the present invention, it was confirmed that the degree of thickening by UV was relaxed by treating the composition with a mouse irradiated with ultraviolet rays. In addition, by observing the increase and decrease of the gene in the skin tissue using the cDNA microarray, 32 kinds of genes in the skin were reduced by UV. However, 18 genes were increased by treating the composition containing the MAO liposome as an active ingredient And recovered to normal level. Also, clinical studies were conducted to confirm that the composition of the present invention was treated to reduce the amount of transdermal water loss, and it was confirmed that skin erythema was reduced, thereby protecting skin damaged by ultraviolet rays. Thus, it was confirmed that the cosmetic composition or the pharmaceutical composition containing the maleic-grafted liposome of the present invention as an active ingredient is excellent in the image treatment and ultraviolet ray blocking effect.

The cosmetic composition or pharmaceutical composition containing the hemofusogenic liposome of the present invention as an active ingredient provides an excellent effect of treating an image and blocking ultraviolet rays. Also, by treating the composition of the present invention with the skin, the amount of percutaneous water loss can be reduced, and the skin erythema can be reduced, thereby protecting the skin damaged by ultraviolet rays.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic diagram of a formulated inhale liposome formulation according to embodiments of the present invention. FIG.
FIG. 2 is a graph showing the results of particle size analysis of a formulated inosine liposome formulation according to an embodiment of the present invention.
FIG. 3 is a photograph of a lotion, cream and spray type cosmetic composition containing an oil-in-fat liposome as an active ingredient.
4 is a photograph of SKH-1 mice before and after irradiating UVB (302 nm) at 180 mJ / cm ² (15 minutes) according to an embodiment of the present invention.
FIG. 5 is a graph showing the result of H & E staining after immersing and fixing mouse skin tissue (2 x ² cm ² ) according to the embodiment of the present invention in 10% formalin (A: UV untreated control group; C: Care-treatment group after UV irradiation, D: Hemp cream treatment group after UV irradiation, E: Hemp treatment group after UV irradiation, and F: Hemp spray treatment group after UV irradiation).
FIG. 6 shows 1) migration pattern and 2) peak pattern of RNA isolated from mouse skin tissue according to an embodiment of the present invention. FIG.
Figure 7 is a photograph of prototype Spray Type A and Spray Type B for clinical trial.
8 is a schematic diagram of the Tewameter 占 TM300 used to measure the transdermal water loss and the temperature and humidity of the probe in the Tewameter 占 TM300. (Right) in which the amount of evaporation (g / h < 2 >) is measured.
FIG. 9 is a graph showing a change in the transdermal water loss of a subject according to an embodiment of the present invention (Control: negative control group, Test: horsepowder group).
10 is a schematic diagram (left) of a spectrophotometer CM-2500d used to measure skin erythema and a color scheme (right) of CIE (Commission Internationale de l'Eclairage). The brightness is L *, the color is a *, and the saturation is b *).
FIG. 11 is a diagram showing a change in skin erythema of a subject according to an embodiment of the present invention (Control: negative control group, Test: horse coating group).
12 is a photograph of before and after application of a prototype for a clinical trial to a subject according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

Example 1. Manufacture of Mauze inclusion-forming liposome formulation

The raw material for preparing the liposome formulation was adjusted to 100% by adding purified water to hemp (20% by weight) + lecithin (3% by weight) + emulsifying aid (1.3% by weight).

To prepare the liposome formulations, Maehu was melted at 40-45 캜. Lecithin and an emulsifying aid were mixed with the melted horse meat oil and purified water, and dissolved at 65-70 캜.

Thereafter, the above-mentioned MEA mixed solution was put into a microfluidizer. The emulsification was performed by setting the emulsification times to 3-pass, 4-pass and 5-pass by setting the mixture ratio of 20% by weight, 25% by weight and 30% by weight and setting the emulsion pressure to 25,000 psi Table 1).

Liposome M1 Liposome M2 Liposome M3 Moisture content (% by weight) 20 25 30 The number of emulsions (pass) 3 4 5 Pressure Setting 25,000 psi 25,000 psi 25,000 psi

Example  1-1. Mayu Enclosure Liposome  Particle size analysis of formulations

The particle size of the prepared male oil liposome formulation was analyzed. The particle size distribution and peak were confirmed using a particle analyzer (Fig. 2). Temperature 25 ± 2 ° C, relative humidity 67 ± 5% R.H. Environment; The results of the analysis using the laser diffraction method are shown in Table 2 below.

Liposome M1 Liposome M2 Liposome M3 Average particle size
(Mean) 0.43 0.37 0.31

(Unit: 占 퐉)

Examples 1-2. Fatty acid analysis

Fatty acid analysis was performed to identify unsaturated fatty acids that were not encapsulated in the phospholipids after the preparation of the liposome of the present invention. The test was carried out according to the method described in Article 10 1.1.5.4. The test environment was set at a temperature of 26 ± 2 ° C and a relative humidity of 67 ± 2% R.H. The results are shown in Table 3.

Ji-Bin Mountain Liposome M1 Liposome M2 Liposome M3 Mayu 100% C12: 0 Lauric Acid 0.000 0.000 0.000 10.131 C14: 0 Myristic Acid 0.210 0.095 0.000 1.389 C14: 1 Myristicoleic Acid 0.047 0.000 0.000 0.169 C15: 0 Pentadecanoic Acid 0.000 0.000 0.000 0.169 C16: 0 Palmitic Acid 0.862 0.367 0.099 5.793 C16: 1 Palmitoleic Acid 0.639 0.253 0.062 4.729 C17: 0 Hepadecanoic Acid 0.000 0.000 0.000 0.193 C17: 1 cis-10-Hepadecanoic Acid 0.000 0.000 0.000 0.240 C18: 0 Stearic Acid 0.290 0.149 0.079 1.475 C18: 1n9t Elaidic Acid 0.768 0.306 0.000 5.536 C18: 1n9c Oleic Acid 1.421 0.547 0.099 10.794 C18: 2n6t Linoleaidic Acid 0.056 0.000 0.000 0.346 C18: 2n6c Linoleic Acid 0.287 0.117 0.000 2.007 C20: 1 cis-11-Eicosenoic Acid 0.050 0.000 0.000 0.318 C20: 2 cis-11,14-Eicosadienoic acid 0.000 0.000 0.000 0.053 Sum 4.630 1.834 0.339 43.435

Example  2. Prototype manufacturing

Example  2-1. Manufacture of prototypes for animal experiments

The prototype of the cosmetic composition containing the liposome as an active ingredient was prepared as shown in Table 4 below, and three types (spray type, lotion type, and cream type) were prepared by varying the amount of the thickening agent in the liposome formulations. (Fig. 3).

Raw material name Composition (% by weight) Liposomal hemp (20% by weight) One Mayu 20,000 2 lecithin 2.400 3 ARASEL-165 0.800 4 ARASEL-60 0.500 5 glycerin 3.000 6 1,3 BG (Butylene Glycol (butylene glycol) 7.000 7 Dimethicone 1,000 8 Purified water 62.790 sub Total 97.490 SEPIPLUS-S - Tocopherol 0.100 Ethylenediaminetetraacetic acid (EDTA) 0.010 Allantoin 0.050 Cremophor RH 60 0.200 antiseptic

CS-Bio - Bioran NP200 - 1,2-Hexanediol 2.000 Spices (Herb Complex-9) 0.200 sum 100.00

Example  2-2. Manufacture of prototypes for clinical trials

Prototype for Clinical Trial Spray Type A and Spray Type B were made with the following composition (Table 5).

Raw material name Spray Type A Spray Type B weight% g weight% g Liposomal hemp (20% by weight) One Mayu 20,000 180.00 - - 2 lecithin 2.400 21.60 2.400 21.60 3 ARASEL-165 0.800 7.20 0.800 7.20 4 ARASEL-60 0.500 4.50 0.500 4.50 5 glycerin 3.000 27.00 3.000 27.00 6 1,3 BG (Butylene Glycol (butylene glycol) 7.000 63.00 7.000 63.00 7 Dimethicone 1,000 9.00 1,000 9.00 8 Purified water 62.790 565.11 82.790 745.11 sub Total 97.490 877.41 97.490 877.41 SEPIPLUS-S - - - - Sheer butter - - - - Cetearyl alcohol - - - - Tocopherol 0.100 0.90 0.100 0.90 Ethylenediaminetetraacetic acid (EDTA) 0.010 0.09 0.010 0.09 Allantoin 0.050 0.45 0.050 0.45 Cremophor RH 60 0.200 1.80 0.200 1.80 antiseptic

CS-Bio - - - - Bioran NP200 - - - - 1,2-Hexanediol 2.000 18.00 2.000 18.00 Spices (Herb Complex-9) 0.200 1.80 0.200 1.80 sum 100.00 900.00 100.00 900.00

- A-type: Spray-type prototype with Liposome Mare (50EA manufactured)

- B-type: Spray-type prototype (except for Liposome)

Experimental Example  1. Animal experiments

Experimental Example 1-1. Ensure animal skin model and UV irradiation

SKH-1 hairless mice were introduced from Orient Bio Co., Ltd. The experimental animals were 8 weeks old (female; about 25g body weight) of SKH-1 mice. SKH-1 mice have no hairs and are widely used in skin experiments.

The UV irradiation apparatus was UVB (UVM-225D manufactured by Mineralight, Upland, CA, USA) and UVB (wavelength 302 nm, 50 Watt) was irradiated. First, the light amount of UVB (302 nm) was measured using a photo-Radiometer (HD2102 by Deltaohm, Selvazzano, Italy) with a UV sensor. As a result of the measurement, it was calculated that the irradiation amount required for this experiment is about 15 minutes when the light amount is 180 mJ / cm 2.

UVB (302 nm) calculated above was irradiated with SKH-1 mouse at a dose of 180 mJ / cm ² (15 minutes irradiation) once a day for 7 times in total. As a result, mouse skin was exposed to UV (FIG. 4).

Experimental Example  1-2. Animal Target Mayu Enclosure Liposomes  Active ingredient Cosmetics  Application of the composition

Experimental Example 1-2-1. Separation of experimental group

The UV-irradiated mouse in Experimental Example 1-1 was divided into 6 groups as shown in Table 6 below, and the experiment was conducted. Specifically, 100 mg of each of the animal test prototypes (cream, lotion, spray) (experimental group) and care burn (Korean FAVIS Pharmaceutical Co., Ltd.) (positive control group) of Example 2-1 was applied for three days.

Group Marie UV Processing contents One (-) Control 9 - No treatment 2 (+) Control 9 + No treatment after UV irradiation 3 Hemp cream 9 + Treated hemp cream (100mg) for 3 days after UV irradiation 4 Mayu Lotion 9 + Treatment with Maey Lotion (100mg) for 3 days after UV irradiation 5 spray 9 + 3 days after UV irradiation Maew spray treatment 6 care burn 9 + Treatment with Care Burn (100 mg) for 3 days after UV irradiation

Experimental Example  1-2-2. Autopsy and removal of skin tissue

After the application of the test sample and the care point according to the above Example 1-2-1, observations immediately before the autopsy revealed that the healing of the image was significantly progressed in all the groups. For autopsy, the mice of each group were placed in a CO ² bomb and discarded, and the skin tissue was cut into a size of about 2 × 4 cm ² and divided into a half (2 × ² cm ² ). And half of each mouse skin tissue was used for histology and the other half was used for cDNA microarray.

Experimental Example 1-2-3. Tissue examination

The skin tissue (2 x 2 cm ² ) of Example 1-2-2 was immersed in 10% formalin and fixed, followed by H & E staining (Fig. 5). The results are shown in Table 7.

Treated group Histological findings Remarks A No UV Normal thin skin Normal finding B UV Only UV-thickened cuticle Typical UV symptoms C UV + care burn Reduced degree of thickening by UV D UV + hemp cream Reduced degree of thickening by UV E UV + Mayu Lotion Reduced degree of thickening by UV F UV + Hemp Spray Reduced degree of thickening by UV

Experimental Example  1-3. Increase / decrease analysis of gene expression

In order to observe the effect of the treatment with UVA and the effect of ultraviolet ray blocking on the composition containing the male oil inoculation liposome through the gene expression increase and decrease analysis, the increase and decrease of the genes in the skin tissue of the mice treated with Maul were analyzed using a cDNA microarray.

Experimental Example  1-3-1. Total RNA isolation

Mouse skin tissue (2 x ² cm ² ) was extracted and immersed in RNA later to isolate total RNA. The total RNA quality control (QC) and migration pattern were observed. The migration pattern was observed using an electrophoretic trace, and the peak pattern was observed using an electropherogram (FIG. 6). Table 8 shows the total RNA QC results.

Sample μg / μl OD _{260 / 280} OD _260/230 Total (μg) Ratio (28s / 18s) RIN Result One 0.5832 2.03 1.73 50.1586 1.3 8.9 Check 2 0.9505 1.90 1.74 63.6808 1.9 8.7 Pass 3 0.9440 2.01 2.01 67.0205 1.1 7.2 Check 4 1.0415 2.02 2.00 82.2777 1.0 7.1 Check 5 1.5944 2.01 2.07 122.7703 1.0 7.6 Check 6 1.7311 2.02 2.07 138.4904 1.0 6.9 Check

Experimental Example  1-4. cDNA Microarray

The differences in skin gene expression between the composition-treated and non-treated groups containing the male oil-in-blended liposome were compared and analyzed. In addition, pathway analysis (KEGG DB-based Pathway Analysis (http://www.genome.jp/kegg/tool) was used to investigate the mechanism of burn treatment.

Experimental Example 1-4-1. cDNA Microarray Results # 1

(C / A) (positive (C / A)) test (B / A), but not in the skin Control group) treatment.

GeneSymbol B / A F / A C / A Gene Description Normalized Normalized Normalized Col4a3 0.307442 0.8875373 0.8763529 collagen, type IV, alpha 3 (Col4a3) Ptk2 0.3299705 0.936016 1.0953074 PTK2 protein tyrosine kinase 2 (Ptk2), Krt84 0.3396395 0.6185958 0.8446364 keratin 84 (Krt84) Traf3ip1 0.3544991 0.7279193 0.7912183 TRAF3 interacting protein 1 (Traf3ip1) Fgf12 0.3567277 0.689356 0.6328826 fibroblast growth factor 12 (Fgf12), transcript Fmn2 0.3880981 0.8185694 0.9518416 form 2 (Fmn2) Plekhh2 0.392791 0.7378172 0.8491111 pleckstrin homology domain containing, family Triobp 0.4012961 1.509945 1.0841793 TRIO and F-actin binding protein (Triobp), Tekt1 0.423074 0.7929325 0.9951106 tekt 1 (Tekt1) Fgf7 0.4424168 0.8718929 0.8581263 fibroblast growth factor 7 (Fgf7) Cdh10 0.4487497 0.9331847 1.0725499 cadherin 10 (Cdh10) Dnm1 0.4506292 1.2463303 1.1049801 dynamin 1 (Dnm1) Ctnnd2 0.4527876 0.8446345 0.813263 catenin (cadherin associated protein), delta 2 Reck 0.4612312 0.7051831 0.7632982 reversion-inducing-cysteine-rich protein with accidental motifs Capn3 0.4619306 0.9251997 0.8265232 calpain 3 (Capn 3), transcript variant c Csf1 0.4662032 0.727822 0.8217082 Mus musculus colony stimulating factor 1 Sos1 0.4719723 1.0200799 0.7642666 son of sevenless homolog 1 (Drosophila) Sgce 0.4736147 0.7891463 0.8081125 sarcoglycan, epsilon (Sgce), Arnt 0.4782667 1.0720178 1.0825113 aryl hydrocarbon receptor nuclear translocator Rem1 0.4804995 0.7474961 0.9551005 rad and gem related GTP binding protein 1 Trim67 0.4808753 0.9604717 1.1163534 tripartite motif-containing 67 (Trim67) Krt20 0.4814702 0.7953774 0.9006174 keratin 20 (Krt20), mRNA [NM_023256] Il6 0.4832273 0.720848 0.6101682 interleukin 6 (Il6) Gypc 0.4861633 0.8554984 0.8901 glycophorin C (Gypc) Csf1 0.4866361 0.7202922 0.8166521 colony stimulating factor 1 (macrophage) Adamtsl1 0.4869315 0.7644084 0.8299229 ADAMTS-like 1 (Adamtsl1 Krt39 0.4901856 1.5583493 0.7418905 keratin 39 (Krt39) Dab2 0.4968649 0.7626805 0.771666 disabled homolog 2 (Drosophila) (Dab2), Pgr 0.498142 0.8462876 1.0186397 progesterone receptor (Pgr) Thy1 0.4987533 0.7825556 0.7061281 thymus cell antigen 1, theta (Thy1) Ctnnd2 0.4990211 0.8518003 0.7867853 catenin (cadherin associated protein), delta 2

Experimental Example  1-4-2. cDNA microarray results # 2

(18) Cytoskeleton genes in the following Table 10 were increased in the skin by UV (B / A), but not in the MAO spray (F / A) (test group) Control group) treatment.

GeneSymbol B / A F / A C / A Description Normalized Normalized Normalized Gas2l1 2.0081027 1.216523 1.4571958 growth arrest-specific 2 like 1 Brs3 2.0450335 1.3948373 1.3816317 bombesin-like receptor 3 (Brs3), Kif13b 2.1069632 1.3468128 1.3708618 kinesin family member 13B (Kif13b), Reck 2.1631906 1.3100321 0.6757813 reversion-inducing-cysteine-rich protein Pclo 2.1639616 1.3948373 1.3816317 piccolo (presynaptic cytomatrix protein) Mapk14 2.25682 1.505969 1.3757304 mitogen-activated protein kinase 14 Col4a4 2.2880728 1.0345963 0.9903284 collagen, type IV, alpha 4 Spag16 2.3349123 1.3948373 1.3816317 sperm associated antigen 16 Upk3a 2.3374875 1.3506184 1.6330707 uroplakin 3A (Upk3a), Cckar 2.4030623 1.3948373 1.3816317 cholecystokinin A receptor Pdc 2.4128177 1.3948373 1.3816317 phosducin (Pdc), Fgf1 2.676234 1.3447868 1.74182 fibroblast growth factor 1 (Fgf1), Frmpd4 3.4804137 0.6551608 1.1993526 FERM and PDZ domain containing 4 Cit 3.5012143 1.0210534 1.1184871 citron (Cit), Mapk10 5.332256 1.0077133 1.359891 mitogen-activated protein kinase 10 Frmpd3 5.3386908 0.7735153 0.580281 FERM and PDZ domain containing 3 Cdk5r2 6.1020827 1.4374362 1.5926917 cyclin-dependent kinase 5, regulatory subunit 2

Experimental Example  1-4-3. cDNA Microarray  Results # 3

After UV irradiation, 40 signal transduction genes were reduced in the skin (2/1), but the MAU spray (5/1) (test group) or Careburn (6/1) (positive control) treatment (Table 11). In addition, the expression level of the gene was determined by the cDNA microarray. In addition, signal transduction genes of 38 kinds in the skin different from those of the above 40 signal transduction genes were reduced (2/1), Mayu spray (5/1) (test group) or Carebane (6 / 1) (positive control group), which were recovered to normal levels (Table 12).

2/1 5/1 6/1 GeneS Normalized Normalized Normalized Description Uts2 0.0745054 0.3022452 0.5917122 Urotensin 2 (Uts2) Adcy10 0.0849143 0.150191 0.1079037 adenylate cyclase 10 (Adcy10) Il9r 0.138372 0.5114465 0.372912 interleukin 9 receptor (Il9r), transcript Arhgap36 0.1432571 0.5213844 0.1596192 Rho GTPase activating protein 36 Hmox1 0.1486441 1.0788869 1.1995921 heme oxygenase (decycling) 1 (Hmox1) Angpt4 0.1720192 0.4261671 0.6896707 angiopoietin 4 (Angpt4) Pgr 0.1748606 0.671022 0.7976604 progesterone receptor (Pgr) Iqsec3 0.2235445 0.5017988 0.4115678 IQ motif and Sec7 domain 3 (Iqsec3) Rgs8 0.258555 0.5854728 0.8262045 regulator of G-protein signaling 8 Kir3dl1 0.2773431 0.4091862 0.3377869 killer cell immunoglobulin-like receptor, Kcnip1 0.2817583 0.6464942 0.5884276 Kv channel-interacting protein 1 Capn8 0.2893953 0.6096306 0.4945717 calpain 8 (Capn8), transcript variant 1 Spock1 0.2929815 0.9185698 0.658933 sparc / osteonectin, cwcv and kazal-like Ptpn5 0.3006851 0.9924318 1.0737114 protein tyrosine phosphatase, Chst8 0.3042873 0.8687558 0.9267471 carbohydrate sulfotransferase 8 (Chst8) Adcy1 0.3065195 0.5591779 0.4972752 adenylate cyclase 1 (Adcy1) Lyve1 0.3070753 0.5384592 1.0539024 lymphatic vessel endothelial hyaluronan Ptk2 0.3299705 0.936016 1.0953074 PTK2 protein tyrosine kinase 2 (Ptk2), Arnt2 0.3349901 0.7571333 0.4689128 aryl hydrocarbon receptor nuclear Il17a 0.3374994 0.7219943 0.6350777 interleukin 17A (Il17a) Greb1 0.3386654 0.9684987 0.9706616 gene regulated by estrogen Il11 0.3404994 1.1449553 1.1521395 interleukin 11 (Il11) Ngfr 0.3442054 0.844812 0.8359903 나테스트 자양 반응기 반응기 Cpa6 0.3494969 0.7143385 0.5538527 carboxypeptidase A6 (Cpa6) Wisp2 0.3497745 0.7654958 0.6994343 WNT1 inducible signaling pathway prot Cxcl1 0.3511886 0.7442033 0.5699413 chemokine (C-X-C motif) ligand 1 Fgf12 0.3567277 0.689356 0.6328826 fibroblast growth factor 12 (Fgf12), Stac3 0.3574978 0.9504336 1.173307 SH3 and cysteine rich domain 3 Epha1 0.3662055 0.7780734 1.1090637 Eph receptor A1 (Epha1), Ephb1 0.3718101 0.6062921 0.5161842 Eph receptor B1 (Ephb1), Xcl1 0.3718716 0.6480455 0.5764412 chemokine (C motif) ligand 1 Tnfsf18 0.3859758 0.901809 0.4612663 tumor necrosis factor (ligand) P2ry12 0.3861308 0.7325807 0.8202865 purinergic receptor P2Y, G-protein Fmn2 0.3880981 0.8185694 0.9518416 form 2 (Fmn2) Chrna4 0.391828 0.8307213 0.6387793 cholinergic receptor, nicotinic, Cxcl14 0.3920987 0.7806791 0.6525384 chemokine (C-X-C motif) ligand 14 Rtkn2 0.3952091 0.8421747 0.8056114 rho hectin 2 (Rtkn2) Tff3 0.3954655 0.8507683 0.4226414 trefoil factor 3, intestinal (Tff3) Ednrb 0.4045891 0.6928465 0.9390531 endothelin receptor type B (Ednrb), Asb17 0.4115662 1.5341318 1.5451849 ankyrin repeat and SOCS

2/1 5/1 6/1 GeneS Normalized Normalized Normalized Description Ahrr 0.4142317 0.6631007 0.6378458 aryl-hydrocarbon receptor repressor Clca1 0.4150974 0.7376716 0.7762097 chloride channel calcium activated 1 Smad9 0.4156241 0.7638682 0.8159168 MAD homolog 9 (Drosophila) (Smad9) Gnat1 0.4192503 1.1793123 1.1675428 guanine nucleotide binding protein, NrOb2 0.4210503 0.9477627 1.1215651 nuclear receptor subfamily 0, group B, Spry4 0.4298538 0.7212262 0.642459 sprouty homolog 4 (Drosophila) Cnr1 0.4325572 0.7113653 0.7391145 cannabinoid receptor 1 (brain) Sult6b1 0.4348157 0.949582 0.9473753 sulfotransferase family, cytosolic, 6B, Fgf7 0.4424168 0.8718929 0.8581263 fibroblast growth factor 7 (Fgf7) Muc1 0.4424316 1.3616369 1.1648384 mucin 1, transmembrane (Muc1) Rgnef 0.445084 0.7125061 0.648444 Rho-guanine nucleotide exchange factor Ccl7 0.4462213 1.101274 0.8333874 chemokine (C-C motif) ligand 7 (CCl7) Tro 0.448299 1.2955817 1.3196306 trophinin (Tro), transcript variant 3 Dlx1 0.4504868 0.9282042 0.8914242 distal-less homeobox 1 Dnm1 0.4506292 1.2463303 1.1049801 dynamin 1 (Dnm1) Ctnnd2 0.4527876 0.8446345 0.813263 catenin (cadherin associated protein), Reck 0.4612312 0.7051831 0.7632982 reversion-inducing-cysteine-rich protein Greb1 0.4616207 1.0187886 1.1157739 gene regulated by estrogen Capn3 0.4619306 0.9251997 0.8265232 calpain 3 (Capn 3), transcript variant c Mtf1 0.4642779 1.0526524 0.6448285 금속 응답 faktörü Csf1 0.4662032 0.727822 0.8217082 colony stimulating factor 1 (macrophage) Sos1 0.4719723 1.0200799 0.7642666 son of sevenless homolog 1 (Drosophila) Pde5a 0.4733965 1.4832379 3.358038 phosphodiesterase 5A, cGMP-specific Arnt 0.4782667 1.0720178 1.0825113 aryl hydrocarbon receptor nuclear transloc Gas6 0.4787289 0.9168903 0.9950351 growth arrest specific 6 (Gas6) Mycn 0.4801803 1.0594051 0.8088514 v-myc myelocytomatosis viral related Rem1 0.4804995 0.7474961 0.9551005 rad and gem related GTP binding protein Ntf3 0.4815683 0.7430823 0.8806007 neurotrophin 3 (Ntf3), transcript variant 1 Klrd1 0.4848409 1.045116 0.9734436 killer cell lectin-like receptor, subfamily D, Il1r1 0.4859403 0.8902516 0.7804716 interleukin 1 receptor, type I (Il1r1), Psd3 0.4915959 0.8756355 0.9190251 pleckstrin and Sec7 domain containing 3 Cacna1a 0.4917514 1.017278 0.8755569 calcium channel, voltage-dependent, Tub 0.4932343 0.8754152 0.8375273 tubby candidate gene (Tub) Pcdh7 0.4937756 0.7594658 0.7594658 protocadherin 7 (Pcdh7), Cd55 0.4966334 0.8234972 0.7265639 CD55 antigen (Cd55) Pgr 0.498142 0.8462876 1.0186397 progesterone receptor (Pgr) Ctnnd2 0.4990211 0.8518003 0.7867853 catenin (cadherin associated protein), Il21r 0.499779 1.0768751 1.0078399 interleukin 21 receptor (Il21r)

Experimental Example  1-4-4. cDNA Microarray  Results # 4

After UV irradiation, 44 signal transduction genes were increased in the skin (2/1), but Maou spray (5/1) (test group) or Careburn (6/1) (positive control group) treatment (Table 13). In addition, the cDNA microarray revealed that the cells recovered to the normal level by the microarray.

2/1 5/1 6/1 Genes Normalized Normalized Normalized Description Rps6ka6 2.004398 1.3427523 1.8074757 ribosomal protein S6 kinase Ccbp2 2.048598 0.8455482 0.9782891 chemokine binding protein 2 Hist1h4d 2.0666304 1.0467122 1.109185 histone cluster 1, H4d Hist1h4k 2.0737896 1.0339879 1.129812 histone cluster 1, H4k Hist1h4j 2.091072 0.9439761 1.0973033 histone cluster 1, H4j Kif13b 2.1069632 1.3468128 1.3708618 kinesin family member 13B Apbb1 2.1097834 0.5687836 0.6464117 amyloid beta (A4) precursor protein-binding, Reck 2.1631906 1.3100321 0.6757813 reversion-inducing-cysteine-rich protein Rab39b 2.2742662 1.3948373 1.3816317 RAB39B, member RAS oncogene family Tspan1 2.3120096 0.9061301 4.126893 tetraspanin 1 (Tspan1) Upk3a 2.3374875 1.3506184 1.6330707 Uroplakin 3A (Upk3a) Mpo 2.360833 1.5836024 0.6995787 myeloperoxidase (Mpo), Ntrk2 2.3610423 0.8868418 1.0938727 neurotrophic tyrosine kinase, Sst 2.362114 1.0155731 1.1187074 somatostatin (Sst) Slc26a10 2.3674176 1.2876745 1.4432106 solute carrier family 26, member 10 Sox8 2.375807 1.3948373 1.662846 SRY-box containing gene 8 (Sox8) Pdc 2.4128177 1.3948373 1.3816317 phosducin (Pdc), transcript variant 1 Odz3 2.4468668 1.328273 1.8107141 odd Oz / ten-m homolog 3 (Drosophila) Cntnap2 2.5099263 1.3011103 2.4372687 contactin associated protein-like 2 (nt 1 Guca1b 2.5130641 1.3278861 1.5695499 guanylate cyclase activator 1B (Guca1b) Fgf1 2.676234 1.3447868 1.74182 fibroblast growth factor 1 (Fgf1) Cr2 2.88554 1.5363555 1.3600067 complement receptor 2 (Cr2) Eras 2.9824872 1.3948373 1.3816317 ES cell-expressed Ras (Eras) Rxfp2 3.020863 1.3948373 1.3816317 relaxin / insulin-like family peptide receptor 2 Wnt8b 3.0362053 1.3423874 1.5245091 wingless related MMTV integration site 8b Chrnb3 3.1113834 1.3948373 1.3816317 cholinergic receptor, nicotinic, Caskin1 3.171372 1.1529399 0.9425277 CASK interacting protein 1 (Caskin1) Cit 3.5012143 1.0210534 1.1184871 citron (Cit) Rgs17 4.188015 2.164923 1.8927606 regulator of G-protein signaling 17 (Rgs17), Pcdh15 4.4001656 1.3948373 2.9624395 protocadherin 15 (Pcdh15), transcript variant Mapk10 5.332256 1.0077133 1.359891 mitogen-activated protein kinase 10 (Mapk10), Frmpd3 5.3386908 0.7735153 0.580281 FERM and PDZ domain containing 3 (Frmpd3) Prph2 5.383916 1.3948373 1.3816317 peripherin 2 (Prph2) Gipr 6.0326095 1.6040171 1.0442554 gastric inhibitory polypeptide receptor (Gipr) Mapk8 6.1564355 2.346544 2.292521 mitogen-activated protein kinase 8 (Mapk8) Csf3 6.398417 0.580387 1.3237795 colony stimulating factor 3 (granulocyte) Lrp2 6.792661 1.3948373 1.6839598 low density lipoprotein receptor-related Csn1s1 8.01872 1.5283742 2.060832 casein alpha s1 (Csn1s1) Spock3 10.544639 0.5982416 0.5925778 sparc / osteonectin, cwcv and kazal-like domain CnH3 11.174891 1.8540658 0.9229071 cornichon homolog 3 (Drosophila) (CnH3), Tnfrsf8 15.650518 2.0356605 3.1225863 tumor necrosis factor receptor superfamily, Tpte 21.904913 1.3948373 1.3816317 transmembrane phosphatase Grin1 34.936157 2.249461 1.2052885 glutamate receptor, ionotropic, NMDA1 Chrm1 37.251465 1.947789 2.1437995 cholinergic receptor, muscarinic 1, CNS 1

Pathway analysis confirmed that MAU restored the expression of genes that were increased or decreased in UV-damaged skin cells to normal levels, which was attributed to the effect of restoring the skin barrier, Respectively.

Experimental Example  2. Clinical Trial

Experimental Example  2-1. Subject selection

The subjects of this study were 20 female subjects (mean age 40.1 ± 6.4 years) aged 25 to 48 years with normal skin.

① Subject selection criteria

Applicants who meet the conditions described below were selected as test subjects.

Ⅰ) Healthy adult men and women between the ages of 18 and 60

Ⅱ) Healthy person who does not have grade or chronic physical disease including skin disease

Iii) Applicants who have fully explained the purpose of the examination, contents, etc. and voluntarily signed the examination agreement

② Subject exclusion criteria

Applicants who fall under the following categories are excluded from the test subjects.

Ⅰ) If you plan to become pregnant during pregnancy, lactation, or within 6 months.

Ⅱ) When a skin contour containing steroids is used for more than one month to treat skin disease

Iii) If 3 months have not elapsed after participating in the same test

Iv) If you have sensitive, irritable skin

Ⅴ) If there is a skin abnormality such as dots, acne, erythema, capillary dilatation on the test site

Vi) Using the same or similar cosmetics or medicines in the test site within 3 months of the start of the test

Ⅶ) If you have chronic wasting disease (asthma, diabetes, hypertension, etc.)

Ⅷ) If you have atopic dermatitis

Ⅸ) If it is determined that the test is difficult due to the judgment of the other state tester

③ Restrictions

The subjects were prohibited from mixing with functional cosmetics or drugs other than the prototype.

④ Number of subjects and basis

The number of subjects was 20 or more according to the Guideline for Evaluation of Functional Cosmetics (2003, 2005), Cosmetics Labeling and Test Method Guideline for Advertising Demonstration (2013).

Experimental Example  2-2. Subject's skin characteristics

Twenty one subjects (# 03, failure to induce erythema) were excluded from the study. Twenty of the female subjects aged 25 to 48 years (mean 40.1 ± 6.4 years) We performed diligently the process. The skin characteristics of the subjects were investigated by the questionnaire, and the analysis results are shown in Table 14 below.

Item Classification Frequency (N) Percentage (%) Skin type
(Skin type) Dry 7 35.00 Normal 9 45.00 Oily One 5.00 Dry and oily 3 15.00 Problematic skin (Problematic) 0 0.00 moisture
(Hydration) Sufficient 0 0.00 Normal 13 65.00 Deficient 6 30.00 Oil
(Sebum) Glossy 2 10.00 Normal 15 75.00 Deficiency 3 15.00 Surface condition
(Surface) Smooth 3 15.00 Normal 16 80.00 Rough One 5.00 Skin thickness
(Thickness) Thin 6 30.00 Normal 14 70.00 Thick 0 0.00 UV exposure time
(Duration of UV exposure) Within 1 hour (Less than 1hr) 5 25.00 1-3 hours (1-3 hr) 14 71.00 More than 3hrs One 5.00 Average sleep time per day
(Sleeping hours) Less than 5hrs (less than 5hrs) One 5.00 5-8 hours (5-8 hrs) 16 80.00 8 hours or more (Above 8hrs) 3 15.00 Smoking
(Smocking) No, 18 90.00 Pim (Yes) 2 10.00 Stimulation sensitivity
(Irritability) Yes (Yes) 4 20.00 No 16 80.00 Slow Sensibility
(Stinging) Yes (Yes) 0 0.00 No 20 100.00 Adverse experience
(Adverse reaction) Yes (Yes) 0 0.00 No 20 100.00

Experimental Example  2-3. Human body target Enclosure Liposomes  Active ingredient Cosmetics  Application of the composition

Prototype for clinical trial Spray type A (test group) and spray type B (control group) were prepared with the composition according to example 2-2. The prepared prototype of the clinical trial (FIG. 7) was washed twice a day (morning and evening) for 6 days, and then the test sample was rinsed with an appropriate amount of the prototype at the designated site (inside of the anterior chamber) and lightly rubbed to be completely absorbed.

Experimental Example  2-3-1. Induced skin barrier damage by UV

The minimum erythema dose (MED) of each subject before the test was measured using Multiport UV Solar Simulator® (Solar light company, USA) to induce skin barrier damage. Erythema was induced by irradiating 2 MEDs of ultraviolet (UV) light at 2 sites (test group, control group) with minimum erythema amount.

Experimental Example  2-3-2. Transdermal water  Loss amount TEWL ) evaluation

The transdermal water loss (TEWL) was measured by using an open chamber Tewameter® TM300 (Courage & Khazaka, Germany). Tewameter® TM300 measured the probe's temperature and moisture evaporation (g / h m2) over time per unit area of the measurement site, based on the diffusion principle in the open chamber (Figure 8). In this test, the amount of transdermal water loss of the test site at each evaluation point was measured three times for 60 seconds each, and the mean value was analyzed.

As a result, the transdermal water loss was significantly reduced at all evaluation time points in the test group and at the 3 and 6 day time points in the control group (p <0.05) (Fig. 9, Table 15). Compared with the control group (24.8%), the transdermal water loss tended to decrease compared to the control group (28.83%) (Table 16).

Group Time Mean
(g / hm) SD SEM p-value Decrement (%) Test Before 7.49 1.54 0.34 - - After UV 8.06 1.96 0.44 - - 1D 7.57 2.21 0.49 0.033 * 6.00 ▼ 3D 6.56 1.90 0.42 0.001 * 18.61 ▼ 6D 5.73 1.17 0.26 0.000 * 28.83 ▼ Control Before 7.79 2.03 0.45 - - After UV 7.89 2.21 0.49 - - 1D 7.80 2.70 0.60 0.780 1.12 ▼ 3D 6.88 2.31 0.52 0.031 * 12.79 ▼ 6D 5.94 1.79 0.40 0.002 * 24.71 ▼

Group Time Mean Square F p-value Test vs. Control Before - - - After UV - - - 1D 2.806 0.784 0.387 3D 0.008 0.002 0.961 6D 0.269 0.127 0.726

Experimental Example  2-3-3. Skin erythema rating

Skin erythema was measured using a spectrophotometer CM-2500d (Minolta, Japan). This is a device for measuring the spectral reflectance of an object color, and tristimulus values are measured and calculated by L *, a *, b *, which is a color scheme of CIE (Commission Internationale de l'Eclairage). L * a * b * The lightness in the colorimetric system is expressed by L *, and the chromaticity indicating the color and saturation is expressed by a *, b *. a * and b * denote the direction of the color, a * denotes a red direction, -a * denotes a green direction, b * denotes a yellow direction, and -b * denotes a blue direction. L *, a *, b * indicate the achromatic color as the value is centered, and the chromaticity increases as the opposite (FIG. 10). In the present study, the average value of a * value (erythema) was measured after measuring the left or right elbow area three times at each evaluation time point.

L *: Luminance parameters

a *: Chrominance parameters green-to-red

b *: Chrominance parameters (color factor blue-to-yellow)

As a result, the erythema was significantly reduced at each evaluation point (p <0.05) in the test group and the control group as compared with the point immediately after the barrier injury (FIG. 11, Table 17). Compared with the control group (23.26%), the erythema tended to decrease in the test group (24.26%) (Table 18).

Group Time Mean
(AU) SD SEM p-value Increment (%) Test Before 6.36 1.23 0.27 - - After UV 13.41 2.08 0.46 - - 1D 12.67 2.33 0.52 0.002 * 5.54 ▼ 3D 10.79 2.55 0.57 0.000 * 19.55 ▼ 6D 10.16 1.72 0.38 0.000 * 24.26 ▼ Control Before 6.08 1.22 0.27 - - After UV 12.80 1.72 0.38 - - 1D 12.25 1.91 0.43 0.010 * 4.03 ▼ 3D 10.36 2.08 0.46 0.000 * 19.05 ▼ 6D 9.84 1.58 0.35 0.000 * 23.16 ▼

Group Time Mean Square F p-value Test vs. Control Before - - - After UV - - - 1D 0.737 1.048 0.319 3D 0.673 0.594 0.450 6D 1.670 2.371 0.140

Experimental Example  2-3-4. Skin safety evaluation

Subjective skin irritation and objective skin irritation were evaluated at each evaluation time point after use of the product by interviews and observations by the tester.

As a result, skin adverse reaction was not observed in all subjects (Fig. 12, Table 19).

Symptom 1D 3D 6D Subjective stimulation
(Subjective irritation) Itching 0 0 0 Stinging like a needle piercing 0 0 0 Burning 0 0 0 Prickling 0 0 0 Stiffness 0 0 0 Tightness 0 0 0 Others (Etc.) 0 0 0 Objective stimulus
(Objective irritation) Erythema 0 0 0 Edema 0 0 0 Desquamation 0 0 0 Papule 0 0 0 Others (Etc.) 0 0 0 Number of total abnormal responders
(Total number of subjects) 0 0 0

From the above results, it was confirmed that the cosmetic composition or the pharmaceutical composition containing the hemofusogenic liposome of the present invention as an active ingredient is excellent in the effect of the treatment of burns and ultraviolet ray shielding.

Statistical significance of all data was analyzed using the SPSS® Package Program (IBM, USA). Regularity was verified by Shapiro Wilk, and comparison of post-posterior comparisons and inter-group comparisons were verified using RM ANOVA (p <0.05). The rate of increase / decrease was calculated according to the following formula.

Increase / decrease ratio (%) = {(measured value before use) - (measured value after use)} / measured value before use × 100

Claims (8)

i) melting the marrow at 40 to 45 占 폚;
ii) mixing the melted horse meat oil with lecithin and an emulsifying aid to dissolve in purified water; And
iii) injecting the dissolution mixture of step ii) into a high-pressure emulsifier and emulsifying the emulsion three to five times at a pressure of 15,000 to 30,000 psi to obtain a liposome encapsulated with the emulsion;
&Lt; / RTI &gt; wherein the liposome is a liposome. The method according to claim 1,
Wherein the dissolution mixture in step ii) is contained in an amount of 10 to 40% by weight, lecithin in an amount of 1.0 to 4.0% by weight, an emulsifying aid in an amount of 0.5 to 3.0% by weight, and purified water in an amount of 55 to 85% by weight. The method according to claim 1,
Wherein the grain size of the talc inclusion liposome is 0.25 to 0.5 占 퐉. The emulsion composition of the present invention is prepared by emulsifying a dissolution mixture containing 10 to 40% by weight of lecithin, 1.0 to 4.0% by weight of lecithin, 0.5 to 3.0% by weight of an emulsifying aid and 55 to 85% by weight of purified water with a high pressure emulsifier to a particle size of 0.25 to 0.5 μm As a liposome. A pharmaceutical composition for the treatment of burns or ultraviolet rays comprising the liposome of claim 4 as an active ingredient. A cosmetic composition comprising the liposome of claim 5 as an active ingredient. The cosmetic composition according to claim 6, wherein the cosmetic composition is for burn treatment or ultraviolet ray shielding. The cosmetic composition according to claim 6, wherein the cosmetic composition is a lotion, nutritional lotion, nutritional cream, massage cream, essence, pack, paste, gel, cream, lotion, powder, soap, oil, foundation, wax or spray formulation Cosmetic composition.

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