KR20170031496A

KR20170031496A - Natural liposome, process for the preparation thereof, and cosmetic composition comprising the same

Info

Publication number: KR20170031496A
Application number: KR1020150129128A
Authority: KR
Inventors: 옥민; 윤경훈; 박진철; 이성은; 성혜영; 이진아; 홍우종; 강민정; 박성수; 박노환
Original assignee: (주)미애부생명과학; 옥민
Priority date: 2015-09-11
Filing date: 2015-09-11
Publication date: 2017-03-21
Also published as: KR101797285B1

Abstract

The present invention relates to a natural liposome containing one or more natural emulsifiers, a solvent and a natural preservative, and a liposome constituent material containing a mixture of a gossam extract, a white ginseng extract or a mixture thereof, a process for producing the liposome, to provide.

Description

[0001] The present invention relates to a natural liposome, a process for preparing the same, and a cosmetic composition comprising the natural liposome, a process for the preparation thereof, and a cosmetic composition comprising the same,

The present invention relates to a natural liposome containing a ginseng extract and a white ginseng extract, a process for producing the same, and a cosmetic composition containing the same. More specifically, the present invention relates to a natural liposome having antibacterial, anti- To a cosmetic composition.

Acne is one of the common skin diseases that 6 ~ 8% of dermatologists in Korea have. Acne has occurred mainly in adolescents in the past, but recently it has occurred in various age groups. That is, even in adults, acne is caused by overwork, stress, physiology and pregnancy, digestive organs and endocrine dysfunction, and chemicals such as cosmetics.

According to Park Jung-min et al. (The effect of skin care using acne powder and propolis on acne skin by Park Jung-min, Han Chae-jeong, and Korea Proprietary Research Institute , 2013, 19.6: 1103-1111), the major cause of acne is male androgen (5α-reductase) in the sebaceous glands, increased production of dihydrotestosterone by sebaceous glands, increased inflammation caused by increased proliferation of sebaceous glands ( Propionibacterium acnes ) in the sebaceous glands, Function abnormality, and the like.

The specific causes of acne are as follows: First, sebum is abnormally excessive secretion. Excess sebum secretion is closely related to the male hormone testosterone. Testosterone is converted to dihydrotestosterone (DHT) by 5 alpha-reductase in skin cells. In addition, it is known that sebaceous gland cells enhance the activity of sebaceous glands and stimulate sebum secretion (Rosenfield RL et al., The American Journal of Medicine, 16, pp. 80-88, 1995).

The second case is that the hair follicle wall is over-shaped and the pores are closed. When sebum is produced, it must be released smoothly out of the skin. However, if the pores are clogged due to the overgrowth of the hair follicles, the smooth release of the sebum is suppressed, and the sebaceous stasis in the hair follicles forms the fine cotton dough. This is the early stage of acne.

Third, stagnant sebum in the hair follicles aggravates acne lesions by promoting the growth of propionibacterium acnes, an anaerobic bacterium that resides in the hair follicles. As the acne bacterium grows in the sebaceous glands, triglycerides constituting sebum are converted into free fatty acids and act as irritants or as comedogenic substances. (Sergio Nacht, Cosmetics Toiletries, 101, pp 47-55, 1986; Registered Patent No. 10-1363028).

Most of the treatments developed to treat these acne are chemical products such as benzoyl peroxide, which has a strong antibacterial action against acne bacteria, clindamycin, which is topically applied antibiotics, and erythyromycin. These treatments can resolve inflammation, but when used for a long time, there are side effects such as resistance to acne bacterium, pruritus, malformations and the like.

Although the use of corticosteroids or estrogen may be considered as an acne remedy, the use of such substances is limited because of the adverse effects such as increased body weight, staining, and thrombosis.

In recent years, surgical treatments such as extrusion of cotton fabrics, chemical peeling, physical peeling, laser peeling, and phototherapy have been developed and used. For example, in the case of peeling, skin irritation is given. .

In order to solve these problems, attempts have been made to search for and develop new skin acne related useful substances from natural substances which can be used comfortably for a long time without harming the skin relatively and which are harmless to human body and secured.

For example, plant extracts or herbal medicine extracts have been proposed as cosmetic compositions for alleviating and inhibiting acne. However, until now, it has been developed as a simple emulsion type or a gel type product in which a cosmetic composition is simply added with a plant extract, so that the absorption capacity of the body is low and thus the efficacy of the extract is limited.

In order to solve such a problem, a liposome that can contain both physiologically active ingredients liposoluble or water-soluble can be used so that the physiologically active ingredient can be delivered stably without breakage.

Liposomes are vesicles formed when a phospholipid is suspended in an aqueous solution and are separated from the outer membrane by a membrane composed of a lipid bilayer, and lipids such as cholesterol glycolipids and membrane proteins can be introduced into the membrane. In addition, liposomes are capable of collecting ions, low molecular substances, nucleic acids, proteins, and the like in the contained aqueous solution layer, thereby helping to transport these materials. For example, liposomes that are sensitive and difficult to absorb into the skin can be transported to the liposomes to help them absorb the ingredients without breaking.

However, the liposome has a problem that the formulation is unstable and the collection efficiency is very low. In addition, cholesterol used as a raw material of liposomes has a problem that liposome lipid membrane formation becomes strong, skin permeability of a capturing material is lowered, and release of an active substance is difficult. In addition, solvents, preservatives, and the like used in the production of liposomes may cause skin irritation. Accordingly, various kinds of liposomes have been studied.

In order to solve these problems, it is necessary to develop a natural substance which is safe to the skin and can be used without any side effects even when exposed for a long time, and it is easily absorbed into the body and is more easily involved in transdermal administration, Development of a composition is required.

On the other hand, Patent No. 1196745 discloses a cosmetic extract for alleviating and preventing acne symptoms obtained by solvent extraction of gosam, rhubarb, yellow white and gold.

Patent No. 1196745

Disclosure of the Invention The present invention provides a natural liposome for anti-acne which is easy to be absorbed into the body without causing problems such as skin irritation and has an antibacterial effect, and a cosmetic composition for improving acne comprising the same The purpose.

In order to achieve the above object, an example of the present invention provides a liposome constituent material containing at least one emulsifier, a solvent and a natural preservative, and a natural liposome containing a combination of a goat extract, a white extract or a mixture of the two .

Another example of the present invention provides a cosmetic composition containing the natural liposome.

Another example of the present invention is a method for producing a microorganism, which comprises a first step of mixing at least one kind of emulsifier with distilled water and ultrasonication at 50 to 80 ° C, adding a crude extract or a white ginseng extract or a mixture thereof to the product of the first step, And a third step of adding a distilled water heated to 40 to 60 ° C and a natural preservative to the product of the second step and subjecting the product to ultrasonic treatment to form a liposome. do.

The present invention also provides a skin absorption promoter, an atopic dermatitis remedy comprising the natural liposome.

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a natural liposome useful for prevention and improvement of acne and a cosmetic composition containing the same .

1 shows a process for preparing a natural liposome according to one embodiment of the present invention.
FIG. 2A is a graph showing the results of measuring the skin absorption power of the natural liposomes of Examples 1 to 4 applied with different solvents, FIG. 2B is a graph showing the results of measurement of skin absorption power of the natural liposomes of Examples 5 to 8, to be.
3 is a graph showing the measurement results of the skin absorption power of the natural liposome of Example 1, the mixture of Comparative Example 1, the liposome of Comparative Example 2, and the synthetic liposome of Comparative Example 3.
4A to 4C are SEM photographs of the liposome of Example 1, the mixture of Comparative Example 1, and the liposome of Comparative Example 3, respectively.
Figs. 5A to 5C show particle size distribution diagrams of the liposome of Example 1, the mixture of Comparative Example 1, and the liposome of Comparative Example 3, respectively.
Fig. 6 is a graph of absorbance for cytotoxicity confirmation according to the treatment of the liposome of Example 1, the mixture of Comparative Example 1, and the liposome of Comparative Example 3. Fig.
7 is a graph showing inhibition rates of inflammation-inducing substances according to the treatment of the liposome of Example 1, the mixture of Comparative Example 1, and the liposome of Comparative Example 3. Fig.
FIG. 8 is a graph showing the content of TNF-a according to the treatment of the liposome of Example 1, the mixture of Comparative Example 1, and the liposome of Comparative Example 3. FIG.
FIGS. 9A to 9F show the results of inhibiting activity of erythrocyte agglutination when treated with the mixture of Comparative Example 1, the liposome of Comparative Example 3, the triclosan, and the liposome of Example 1, It is a photograph which shows.
FIGS. 10A and 10B are photographs of the skin of a male mouse after three days of injection before and after the injection of P. acnes, respectively.
Fig. 11 is a photograph of acne lesion area after injecting an acne-causing strain into the back skin of a nude mouse.
12 is a graph showing the DHT content in mouse serum according to the treatment of the liposome of Example 1, the mixture of Comparative Example 1, and the liposome of Comparative Example 3. Fig.
13 is a graph of TNF-a content in mouse serum according to the treatment of the liposome of Example 1, the mixture of Comparative Example 1, and the liposome of Comparative Example 3. Fig.
14 is a graph showing the IL-6 content in mouse serum according to the treatment of the liposome of Example 1, the mixture of Comparative Example 1, and the liposome of Comparative Example 3. Fig.
15 is a graph of the spleen index of a mouse according to the treatment of the liposome of Example 1, the mixture of Comparative Example 1, and the liposome of Comparative Example 3. Fig.
16 is an optical microscope photograph of the ear of a mouse according to the treatment of the liposome of Example 1, the mixture of Comparative Example 1, and the liposome of Comparative Example 3. Fig.

Hereinafter, a natural liposome according to the present invention will be described in detail with reference to the accompanying drawings.

However, these descriptions are provided only to illustrate the present invention, and the scope of the present invention is not limited by these exemplary explanations.

1. Emulsifier

Emulsifiers consist of phospholipids and fatty acids.

Examples of the phospholipid include phosphatidylcholine, lysophosphatidylcholine, phosphatidylethanolamine, palmitic acid, hydrogenated phophatidylcholine (derived from soybean), phosphatidylserine, phosphatidylglycerol at least one phospholipid selected from the group consisting of phosphatidylglycerol, phosphatidylinositol, and their hydrogenation products can be used.

As the fatty acid, at least one fatty acid selected from the group consisting of stearic acid, palmitic acid, oleic acid, linoleic acid and linolenic acid (derived from soybean) is used . The fatty acid may be contained in an amount of 1 to 10% by weight.

As the emulsifier, two or more emulsifiers may be used together. For example, as the first emulsifier, phosphatidylcholine, lysophosphatidylcholine, phosphatidylethanolamine, palmitic acid, stearic acid, oleic acid, linoleic acid, acid and linolenic acid (derived from soybean), and hydrogenated phosphatidylcholine (derived from soybean) may be used as the second emulsifier.

In this case, the amount of the first emulsifier component may be 1 to 5 wt%, and the amount of the second emulsifier component may be 0.5 to 2 wt%.

2. Collecting material

As a capturing substance that can be trapped in the phospholipid layer of the liposome, there can be used a ginseng extract and a white ginseng extract extracted from ethanol, and other herbal medicines such as jiyu oil, rhubarb, licorice, cinnamon, clove and bokbun can be used.

Here, Sophora flavescenes Aiton is a perennial plant with a height of about 1 meter, distributed in Korea, China, Japan and Russia. Gosam is called gosam because it tastes good and its effect is similar to gosan. It has a characteristic odor, persistence and weakness.

The roots contain matrixin (sulfocarbipine), oxymatrine, sophoranol, anagurine, methyltocine, bavitifrin, sapsocarpine, alomatrine and the like as alkaloid components, and as the flavonoid component, Isozynthemol, and the like. The leaves and stems contain ruthelin-7-glucoside.

Gosam is an antimicrobial, anti-inflammatory, anti-allergic, anti-cancer, diuretic, hypoglycemic, antitumor and cardiac muscle strengthening, and has the efficacy of chewing gum (淸热干濕) It is known. In addition, it is known that it can be used for dysentery, subarachnoid, genital pruritus, enteritis, itching of skin.

Bacillus is a medicinal herb of the perennial plant (Bietilla striata R.) belonging to the orchidaceae family. It is native to Korea, Japan and China.

The taste is sweet and sweet. It acts on menopause to strengthen the lungs, stop the bleeding, calm the boogie, and let the newborn grow well. Other effects of hemostasis, stomach ulcer or duodenal ulcer treatment, and antibacterial effect. It is also used for the treatment of the same species, rearing, and burns.

In ShuzoT et al. (ShuzoT, Masae Y, Keiko I. Anti microbial lagents from Bletillastriata . Phytochemistry. 1983; 22 (4): 1011-1015), three new antibacterial bibenzyls and two It has been reported that there is a new dihydrophenanthrene. In Yamaki et al. (Yamaki M, Bai L, Kato T, Inoue K, Takagi S. Three dihydrophenanthropyrans from Bletillastriata . Phytochemistry.1993; 32 (2): 427-430), three new components, bletilol A , B, and C were reported. In SaitoN, KuM, TatsuzawaF, LuT, YokoiM, Shigihara A, Honda T. Acylated cyaniding lycosides in thepurple-red flowers of Bletillastriata.Phytochemistry 1995 (40 (5): 1523-152912) A new cyanidin glycoside was analyzed.

The mixture of gossam extract and white ginseng extract may be mixed at a certain ratio and may be contained in an amount of 1 to 50 wt% of the total volume at the time of preparation. When the above-mentioned collecting material is used as a mixture, it is preferable to use a mixture in which the gossam extract and the white ginseng extract are mixed in a weight ratio of 1: 3 to 3: 1, and in particular, in a weight ratio of 1.5 to 2.5: It is preferable to use a mixture.

3. Solvent

As the solvent, distilled water, butylene glycol, propylene glycol, glycerin, and ethanol may be used. As the solvent, a natural solvent is preferable. For example, in the case of butylene glycol or glycerin, a raw material derived from a natural source is used, and a raw material derived from a natural source such as a fermented alcohol is also used. More preferably, distilled water, glycerin, and most preferably distilled water is used. The content of the solvent is the balance of the content of the emulsifier, the trapping agent and the preservative.

4. Preservatives

As a preservative, one or more natural extracts selected from the group consisting of a superfine fruit extract, a flower leaf extract, and an Earther extract may be used. It is distinguished from benzoic acid, a paraoxybenzoic acid ester, a mixture of methylchloroisothiazolinone, phenoxyethanol and the like, which is a commonly used preservative in that it is a natural extract. The preservative may generally be included in an amount of 1 to 5% by weight.

5. Preparation of raw ginseng extract and white ginseng extract

The red ginseng and white ginseng were immersed in distilled water or alcohol, and they were immersed in the solution for 24 hours at room temperature, filtered to remove the solids, and the filtered extract was recovered. In the case of extracting with alcohol as a solvent, it is more preferable that the extract has better antibacterial activity. As the alcohol, methanol, ethanol, butanol and the like can be used, and it is particularly preferable to use ethanol in that a separate solvent removing step is unnecessary. In the case of using a solvent in which ethanol is diluted to a concentration of 50% in distilled water, the extract is particularly preferable in that it exhibits strong antimicrobial activity.

In case of mixing gosam extract and white ginseng extract, the mixing ratio of ginseng extract and white ginseng extract can be adjusted in the range of 1: 3 to 3: 1. In terms of antibacterial activity, a ratio of 1.5 to 1 to 2.5: 1 is particularly preferable.

Ultrasonic treatment may be applied if necessary to increase the recovery of the active ingredient of the extract. Ultrasonic treatment is carried out at 30 Amp for 10 minutes.

6. Preparation of liposomes

The liposome according to the present invention can be manufactured using ultrasonic waves or using emulsification equipment.

end. How to use ultrasonic waves

One or more emulsifiers are mixed with a part of distilled water and ultrasonicated at 50 to 80 ° C using an ultrasonic liquid processor. Ultrasonic fracturing is an operation to homogenize the mixture. When the emulsifier is homogenized, it is mixed with ultrasonic wave after addition of the capturing material. When the emulsifier and the capturing material are completely homogenized, the remaining distilled water and the natural extract (preservative) warmed to 40 to 60 ° C are added and mixed by ultrasonic disruption to form liposome type particles. Thereafter, the temperature is gradually lowered and ultrasonication is finally performed for 5 minutes to make the particles smaller and uniform so that a natural liposome is finally produced. The manufacturing process is shown in Fig.

I. How to use oil painting equipment

Mix at least one kind of emulsifier with a part of distilled water and mix by using homomixer at 1300 ~ 1500rpm and 50 ~ 80 ℃ for 10 ~ 30 minutes. After the emulsifier is homogenized, the emulsifier is added to the emulsifier and mixed with the emulsifier. When the mixing of the emulsifier and the capturing material is finished, the remaining distilled water and the natural extract are added to prepare the natural liposome. Then the particles are made small with an ultrasonic disintegrator to produce the final natural liposome.

The liposome according to the present invention can increase the skin permeability and improve the absorption rate, so that the active ingredient can effectively act on the skin. The liposome according to the present invention can be used for the treatment of acne, prevention and alleviation of atopic skin. In particular, the liposome according to the present invention can be used in cosmetic compositions for such purposes. For example, softening longevity, convergent lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, body lotion, body cream, body oil, body Cosmetics, essences, makeup base, foundation, hair dye, shampoo, rinse, body cleanser, toothpaste, mask sheet and oral care form composition.

Meanwhile, the liposome according to the present invention can also be used as a therapeutic agent for acne treatment, atopic skin prevention and alleviation.

&Lt; Example 1 >

The liposomes were prepared according to the composition shown in Table 1 below.

Raw material Example 1
(Unit: wt%) Emulsifier Emulsifier 1 2.5 Emulsifier 2 One antiseptic One Solvent
(Emulsification process) Distilled water (D.I.Water) 30 Solvent
(Homogenization process) Distilled water (D.I.Water) Balance Capture material Mixture of 50% Gossam Extract and 50% White Extract 20

Emulsifier 1 Emulsifier 1 Emulsifier 1 A mixture of 67 wt% of phosphatidylcholine, 8 wt% of lysophosphatidylcholine, 8 wt% of phosphatidyl ethanolamine, 3 wt% of palmitic acid, 1 wt% of stearic acid, 3 wt% of oleic acid, 7 wt%, linolenic acid 3 wt%

Emulsifier 2: Hydrogenated phophatidylcholine (derived from soy)

Preservative: Pine nuts fruit extract: Waxy pine extract: Earth nere extract is included in weight ratio of 1: 1: 1

The specific manufacturing process is as follows.

First, as shown in Table 1, Emulsifier 1 and Emulsifier 2 were mixed with a part of distilled water (1/3) and ultrasonically disrupted at 65 ° C using an ultrasonic liquid processor (QSONICA, USA). When the emulsifier was homogenized, the extracts of gosam and white extract were added, and the mixture was sonicated and mixed. When the emulsifier and the collecting material were completely homogenized, distilled water (2/3) heated to 50 ° C and a preservative as a natural extract were mixed and ultrasonically pulverized to form liposome-type particles. Thereafter, the temperature was gradually lowered and ultrasonication was performed for 5 minutes to make the particles smaller and uniform. Finally, a natural liposome containing the extract of Gossam ginseng and the extract of Bacillus was prepared.

&Lt; Examples 2 to 4 >

The natural liposomes of Examples 2, 3 and 4 were prepared in the same manner as in Example 1, except that ethanol, glycerin and propylene glycol were used as solvents, respectively.

&Lt; Examples 5 to 8 >

6, 7 and 8 were obtained in the same manner as in Example 1, except that the content of the extract of Bombyx mori and the extract of Bombyx mori was 10%, 20%, 30% and 40%, respectively, .

&Lt; Comparative Example 1 &

In Comparative Example 1, the gosam extract and white ginseng extract used as the capturing material in Example 1 were diluted with distilled water to the same concentration. That is, the extracts of Gossam ginseng extract and white ginseng extract extracted with 50% ethanol were mixed at a weight ratio of 2: 1, and the extracts of Gossam ginseng and Ginseng extract were used in the same amounts as those contained in the natural liposome of Example 1. This mixture was mixed with distilled water to make the same concentration as that of the natural liposome of Example 1 and used in the experiment.

&Lt; Comparative Example 2 &

In Comparative Example 2, a natural liposome was prepared in the same manner as in Example 1, except that the extract of Gossam and the extract of Bacillus was not contained.

&Lt; Comparative Example 3 &

In Comparative Example 3, a synthetic liposome containing a crude extract, a white extract, lecithin and propylene glycol was prepared by using a conventional technique.

Propylene glycol (30%), which is a synthetic solvent, was used as a solvent, and phospholipids (lecithin-1%) were mixed and heated to 65 ° C. When the phospholipids were completely dissolved, the same amount of the extract as in Example 1 and the white extract were homogenized with a homogenizer. The remaining water was mixed with an antiseptic (paraben 0.02%) to form liposome particles, and the particles were homogenized through ultrasonic treatment.

<Experimental Example 1> Confirmation of skin absorption using Franz Diffusion cell

(1) The natural liposomes of Examples 1 to 4 were subjected to a skin absorption test using a Franz Diffusion cell. The results are shown in Fig.

As a result, as shown in FIG. 2A, when the distilled water (DI WATER) of Example 1 was used, the absorption power was the best, and the propylene glycol of Example 3 and the glycerin of Example 4 were lower in absorption than the extract Respectively.

(2) The natural liposomes of Examples 5 to 8 were subjected to skin absorption test using Franz Diffusion cell. The results are shown in FIG. 2B.

As a result, as shown in FIG. 2B, when the content of the crude extract and white extract of Example 1 was 50%, the absorption power was the best.

(3) On the other hand, the skin absorption test was conducted on the natural liposome of Example 1, the mixture of Comparative Example 1, and the synthetic liposome of Comparative Example 3. The results are shown in Fig.

As a result of the experiment, it was shown that the absorption efficiency of Example 1 was the best as shown in FIG.

<Experimental Example 2> Measurement of collection rate

A certain amount of the natural liposome suspension of Example 1 was taken, and then the non-captured material in the liposome was removed using a 0.45 μm syringe filter. Then, ethanol was used to destroy the liposome membrane. The maximum absorption wavelength of Ganoderma lucidum extract was determined. The measured values were substituted into the following formula to confirm the collection efficiency of the active substance in the liposome, and the results are shown in Table 2 below.

(%) = Concentration of the functional substance passing through the filter / Concentration of the first functional substance * 100

Sample Collection rate (%) Example 1 92.89 Comparative Example 3 89.61

As a result of measurement, it was found that the natural liposome of Example 1 was superior in the collection rate to that of Comparative Example 3. Since the comparative example 1 is a raw material which is not liposomized, the collection rate was not measured.

<Experimental Example 3> Analysis of morphology and size of liposome by scanning electron microscope

The particle shape and size of the natural liposome of Example 1, the mixture of Comparative Example 1 and the synthetic liposome of Comparative Example 3 were analyzed using a scanning electron microscope (Scanning Electron microscope S-1700, Hitachi). The measurement was carried out after each sample was lyophilized at -70 ° C, and then thinly spread and coated once with a PdPt mixed catalyst.

The natural liposome of Example 1, the mixture of Comparative Example 1, and the synthetic liposome of Comparative Example 3 were confirmed by scanning electron microscope and SEM photographs were taken. The results are shown in FIGS. 4A, 4B and 4C, respectively.

As shown, the natural liposome of Example 1 formed relatively circular particles, but the mixture of Comparative Example 1 did not show the particle shape and the liposome of Comparative Example 3 had a larger particle shape than the natural liposome of Example 1 .

<Experimental Example 4> Size analysis of liposome by laser particle size analyzer

The particle sizes of the samples of Example 1, Comparative Example 1 and Comparative Example 3 were measured and the particle size distribution was analyzed using light scattering through a laser particle size analyzer (Electrophoretic Light Scattering Spectrophotometer ELS-8000, Otsuka).

The average particle size is shown in Table 3, and the particle size distributions of the natural liposome of Example 1, the mixture of Comparative Example 1 and the synthetic liposome of Comparative Example 3 are shown in FIGS. 5A, 5B and 5C, respectively.

Sample Average particle size (nm) Example 1 206.7 Comparative Example 1 246.1 Comparative Example 3 914.2

As shown in Table 3, the average particle size of the natural liposome of Example 1 was the smallest at 206.7 nm, and the particle distribution was stable and uniform as shown in FIG. 5A.

<Experimental Example 5> Cytotoxicity test using MTT

1 × 10 ⁵ Fibroblast cells were placed in 48 wells and replaced with fresh medium after 24 hours. Then, the natural liposome of Example 1, the mixture of Comparative Example 1, and the synthetic liposome of Example 3 were mixed at a concentration of 0.1 to 5% Lt; / RTI > Subsequently, 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetra zolium bromide (0.1 mg / ml) was added to each well and incubated for 3 hours. Insoluble formazan formed was dissolved in 0.04 N HCl / isopropanol And absorbance was measured at 570 nm through an ELISA reader. The results of the experiment are shown in Fig.

<Experimental Example 6> Measurement of the inhibitory effect of inflammatory substance (NO) by acne bacteria (p.acnes)

Forty-eight wells were dosed with 1 × 10 ⁵ macrophages, and after 24 hours, the natural liposome of Example 1, the mixture of Comparative Example 1 and the synthetic liposome of Example 3 were treated for 48 hours, respectively. Inflammation was induced by treating acne bacterium p.acnes with 1 mg / ml. After the treatment, the culture supernatant and Griess reagent (Sigma G-4410) solution were mixed 1: 1, reacted for 15 minutes, and absorbance was measured at 540 nm. The amount of NO produced was compared based on the amount of NO produced in P. acnes treated cells. Indomethacin 0.5% and triclosan 0.1% were treated in the same amount as the treatment group, and the mixture of Comparative Example 1, the natural liposome of Example 1 and the synthetic liposome of Comparative Example 3 were treated at the same concentration. The results are shown in Fig.

As shown in FIG. 7, the NO production inhibitory effect was confirmed, and it was found that the anti-inflammatory effect was most excellent by decreasing NO production amount up to 66.37% in the cells treated with Example 1.

Experimental Example 7 Measurement of Inflammatory Cytokine Production by Acne Bacterium (p.acnes)

Forty-eight wells were dosed with 1 × 10 ⁵ macrophages, and after 24 hours, the natural liposome of Example 1, the mixture of Comparative Example 1 and the synthetic liposome of Example 3 were treated for 48 hours, respectively. Inflammation was induced by treating acne bacterium p.acnes with 1 mg / ml. After the treatment, the amount of TNF-α produced in the culture supernatant was measured using an ELISA kit (ALPCO). The results are shown in Fig.

The effect of TNF-α secretion on the acne-treated cells was confirmed. The amount of TNF-α was increased by acne bacteria and the amount of secretion was decreased by positive controls such as indomethacin and triclosan. When the natural liposomes of Example 1 were treated, the amount of TNF-? Production was found to be lower than that of Comparative Examples 1 and 3.

<Experimental Example 8> Inhibition of erythrocyte aggregation reaction by acne bacteria

The cultured P. acnes was diluted with phosphate buffer and mixed with an equal volume of red blood cell solution. The mixture was incubated at room temperature for 30 minutes to 3 hours under mild agitation. In order to confirm the inhibitory activity against aggregation reaction, P. acnes solution, extract, and natural liposome were mixed, reacted at room temperature, mixed with red blood cell solution and reacted, and the coagulation reaction was observed under a microscope. (-) reaction is shown in FIG. 9a, (+) reaction is shown in FIG. 9b, and the experimental results are shown in FIGS. 9c to 9f.

Confirmation activity of erythrocyte aggregation reaction inhibition

As a result of measuring the activity of inhibiting the erythrocyte aggregation reaction by P. acnes, it was found that the treatment of the mixture of Comparative Example 1 and the synthetic liposome of Comparative Example 3 did not inhibit the aggregation of erythrocytes by p.acnes 9c, Fig. 9d). Inhibition activity was observed when triclosan, a positive control, was treated (Fig. 9E). Even when the natural liposome of Example 1 was treated, an inhibitory activity similar to that of triclosan was confirmed in erythrocytes (Fig. 9f).

[Animal experiment]

<Experimental Example 9> Effect of P. acnes-induced hairless mouse on acne

Acne-causing strain

P. acnes (ATCC 6919), a causative organism used for acne induction, was purchased from the Korean microbial strain bank. The strains were incubated at 37 ° C for 48 hours under anaerobic conditions using Reinforced Clostridium Medium (RCM, Oxford, Hampshire).

Experimental animals and acne induced

Hos: HR-1 (hairless mice) was used for the experiment. Five-week-old male mice weighing 20g were received from BioLink (Chungcheongbuk-do, Korea) and treated for 4 weeks It took a while. During the test period, feed and water were freely taken, and the temperature of the breeding room (22 ± 2 ℃), relative humidity (55 ±%) and intensity were maintained for 12 hours.

Acne induction was performed by injecting 20 ul of P. acnes ( ¹ x 10 ⁷ CFUs) into the back of a mouse. As shown in FIG. 10A, acne appeared on the clean skin initially after 3 days as shown in FIG. 10B. From 3 days after the acne was induced, samples were applied to each group to confirm the acne relieving effect.

FIG. 11 shows a photograph of the effect of alleviating the acne lesion area after injecting the acne-causing strain into the back skin of the nude mouse and photographing the result with a digital camera.

In the normal group (P. acnes (-)) without the strain, clean skin was maintained. In the group treated with DW (P. acnes (+)), acne was formed even after 18 days there was.

When the sample was treated, it was confirmed that the acne was gradually alleviated. The degree of acne relief was highest in the natural liposome of Example 1, followed by 0.05% Triclosan used as a positive control, the mixture of Comparative Example 1, and the synthetic liposome of Comparative Example 3, in that order.

<Experimental Example 10> Measurement of DHT content in blood

On the last day of the experiment, the mice were fasted for 12 hours or more and then lightly anesthetized with ether. After the laparotomy, blood was collected from the abdominal heart, left at room temperature for about 15 minutes and centrifuged at 10,000 rpm for 30 minutes at 4,000 rpm. After separation, the DHT content was measured using DHT (dihydrotestosterone) ELISA kit (Enzo). The results are shown in Fig.

5 alpha - reductase activity increased the activity of DHT, and DHT contents in mouse serum were compared.

The results showed that DHT content was the highest at 612.10pg / ml in the group treated with DW only after acne induced. Positive control (PC) and the natural liposome of Example 1 showed 320.85 and 376.15 pg / ml, respectively, in the sample treatment group, and DHT content similar to that of the normal group was shown. From this, the effect of the natural liposome-treated group of Example 1 on the acne relief was confirmed.

Experimental Example 11 Measurement of TNF-a content in blood

To determine the content of TNF-a, a cytokine in the inflammatory response, the secretion level was determined using the separated serum. The experiment was carried out using a TNF-a (mouse) ELISA kit (ALPCO). The experimental results are shown in Fig.

The degree of acne relief was confirmed by confirming the content of TNF-a, one of the cytokines that cause acne.

The TNF-a content was 13 pg / ml in the normal group and 19.267 pg / ml in the DW-treated group. In the positive control Triclosan treated group and the natural liposome treated group of Example 1, the TNF-a content was 14.733 pg / ml and 11.133 pg / ml, respectively.

<Experimental Example 12> Measurement of blood IL-6 content

In order to confirm the content of IL-6, a cytokine in the inflammatory reaction, the secretion amount was confirmed by using the separated serum. The amount of IL-6 secretion was confirmed using Mouse IL-6 ELISA Kit (KOMABIOTECH). The experimental results are shown in Fig.

As shown in FIG. 14, the IL-6 content in the normal group was 1.32 pg / ml, and the IL-6 content in the group treated with distilled water after acne induced was 48.68 pg / ml. In the positive control group, Triclosan treatment group showed 28.43 pg / ml, while in the natural liposome treated group of Example 1, the value was 22.25 pg / ml. Accordingly, it is considered that the treatment of the natural liposome of Example 1 can reduce the inflammatory reaction.

<Experimental Example 13> Confirmation of spleen index

For immunological evaluation, the mice were sacrificed on the last day of the experiment, and the spleen was excised and its weight was measured. The results are shown in FIG.

Spleen levels were obtained by dividing the mouse spleen weight (mg) by the mouse weight (g) at the end of the experiment.

The spleen index was 3.09 mg / g in normal group without acne, but increased to 4.35 mg / g in acne group. In the case of treatment with the natural liposome of Example 1, 3.65 mg / g was the most similar to that of the normal group and the lowest value was observed in the other groups except the normal group. These results suggest that the natural liposome of Example 1 significantly inhibited the increase of T lymphocytes in the spleen.

<Experimental Example 14> Histological evaluation

20ul strains of P. acnes (1x10 ⁷ CFUs), which is an acne-causing strain, were injected intradermally into the ear of a mouse and then coated with triclosan, the natural liposome of Example 1, the mixture of Comparative Example 1 and 200ul of the synthetic liposome of Comparative Example 3, respectively.

Twenty-four hours after the start of the experiment, mice were sacrificed and tissues were harvested. The ear tissues were examined to see how they affect the tissues. The tissues were fixed in 10% formalin, embedded in paraffin, stained with hematoxylin-eosin (H & E), and histologically examined by light microscopy.

The results are shown in Fig.

As a result of the experiment, it was confirmed that acne was induced in the group to which DW was applied, whereas it was confirmed that acne was alleviated in the mixture of Comparative Example 1 and the natural liposome of Example 1.

In addition, H & E stain for the histological observation of the ear tissue showed that the ear tissues were thin in the normal group not infected with acne bacterium but swollen in the ear tissue due to edema in the DW group.

Triclosan (PC) and the synthetic liposome of Comparative Example 3 showed more acne tissue relaxation than DW but no significant difference.

It was confirmed that the natural liposome of Example 1 exhibited the effect of alleviating the acne remarkably similar to that of the normal group.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention. The present invention is not limited to the drawings.

Claims

A liposome constituent material containing at least one natural emulsifier, a solvent and a natural preservative, and
As the capturing substance, the extracts of Gossam extract, White extract or a mixture of the two
&Lt; / RTI >

The method according to claim 1,
Wherein the capturing material is a mixture of the gossam extract and the white ginseng extract in a weight ratio of 1: 3 to 3: 1.

The method according to claim 1,
Wherein the natural emulsifier is selected from the group consisting of phosphatidylcholine, lysophosphatidylcholine, phosphatidylethanolamine, palmitic acid, hydrogeneated phosphatidylcholine (derived from soybean), phosphatidylserine, phosphatidylglycerol, phosphatidylinosylol, A natural liposome comprising more than one species of phospholipids.

The method according to claim 1,
Wherein the natural emulsifier comprises at least one fatty acid selected from the group consisting of palmitic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid (derived from soy).

The method according to claim 1,
Wherein the solvent comprises at least one of distilled water, butylene glycol, propylene glycol, glycerin, ethanol.

The method according to claim 1,
Wherein the natural preservative is at least one natural extract of natural liposomes selected from the group consisting of an extract of Acerola japonica, an extract of Acerola sp.

The method according to claim 1,
2.5 to 17% by weight of a natural emulsifier,
1 to 5% by weight of a natural preservative,
1% to 50% by weight of a mixture of a goat extract and a white extract
Solvent residue
&Lt; / RTI >

The method according to claim 1,
Natural liposome for promoting skin absorption.

The method according to claim 1,
Natural liposome for prevention and relief of atopic dermatitis.

10. A cosmetic composition comprising the natural liposome of any one of claims 1 to 9.

A first step of mixing at least one emulsifier with distilled water and ultrasonic treatment at 50 to 80 캜,
A second step of adding a ginseng extract or a white ginseng extract or a mixture thereof to the product of the first step,
Adding a distilled water warmed at 40 to 60 ° C to the product of the second step and a natural preservative and sonicating to form a liposome;
10. The method according to any one of claims 1 to 9,

12. The method of claim 11,
The fourth step of performing ultrasonic processing while gradually lowering the temperature after the third step
&Lt; / RTI >

A skin absorption promoter comprising the natural liposome of claim 1.

A therapeutic agent for atopic dermatitis comprising the natural liposome of claim 1.