KR102485384B1 - Cosmetic composition with multi lamellar liposome encapsulated with the extract of Adenophora triphylla var. japonica HARA. and the extract of Siegesbeckiae Herba for improvement of acne - Google Patents

Abstract

The present invention relates to a cosmetic composition for improving acne containing multi-lamellar liposomes collected by water extracts obtained by re-extracting water extracts of zandae extract, supercritical fluid extract of Heechum and carbon dioxide supercritical fluid extract of Heechum. The present invention relates to a cosmetic composition for improving acne. By properly managing Propionibacterium Acnes , known as the causative bacteria, an improvement effect on acne lesions can be expected.

Description

Cosmetic composition with multi lamellar liposome encapsulated with the extract of Adenophora triphylla var. japonica HARA. and the extract of Siegesbeckiae Herba for improvement of acne}

The present invention relates to a cosmetic composition for improving acne containing multilamellar liposomes in which extracts of Zanjadae and Heechum extracts are collected, and more particularly, generated during extraction of Zandae extract, Heechum supercritical fluid extract and Heechum carbon dioxide supercritical fluid. It relates to a cosmetic composition for improving acne containing multilamellar liposomes in which a water extract obtained by re-extracting the extract gourd with water is collected.

Recently, interest in skin beauty has increased, and the occurrence of acne can lead to various mental diseases such as psychological burden, loss of confidence, and depression beyond skin diseases. Acne mainly occurs on the face, such as the forehead, but also occurs on the chest, back, and shoulders. Acne is a chronic inflammatory disease of the hair follicle and sebaceous gland characterized by the formation of comedones, papules, pustules, etc., and in the past occurred mainly in adolescent men and women. However, due to air pollution, drug abuse, etc., it has recently occurred regardless of age.

The cause of acne has not yet been clearly identified, but hormone action, excessive sebum secretion, and bacterial infection are believed to be the causes. After all, acne can be defined as a complex inflammatory skin disease that occurs in hair follicles, hair follicles and surrounding tissues due to the above causes, and a rash called comedones is a characteristic symptom. Comedon refers to a screw-shaped lump of hardened sebum that appears mainly on the face, forehead, and bridge of the nose. Inside the comedones, anaerobic Propionibacterium acnes , known as the causative agent of acne, grows, and on the surface of the comedones, aerobic bacteria such as Staphylococcus epidermidis , Pityrosporum ovale and Star Staphylococcus aureus and the like grow, and various components produced by these bacteria settle on the hair follicle and surrounding skin to cause inflammation.

Conventionally, oral or external medications have been used to treat acne. The main targets were exfoliation of pores using salicylic acid or retinoic acid, a derivative of vitamin A, and enhancement of bactericidal power against acne bacteria using antibiotics such as triclosan. However, salicylic acid is classified as a preservative in Korea and cannot contain more than 0.5% in the formulation, and triclosan has a side effect of causing skin irritation when in contact with the skin, so their use is inevitably limited.

Therefore, there is an urgent need to develop a technology that is effective in treating acne while having fewer side effects.

Korean Patent Publication No. 1020130131508 (published on 2013.12.04) describes a composition for preventing and treating acne containing brazilein, an active ingredient of a water extract residue of Caesalpinia sappan L. Korean Patent Publication No. 1020190130340 (published on November 22, 2019) describes a cosmetic composition for improving acne containing an extract of Lycopus lucidus or a fraction thereof.

In the present invention, by properly using antibacterial and anti-inflammatory materials derived from natural products, and properly managing Propionibacterium Acnes , known as acne-causing bacteria, an improvement effect on acne lesions can be expected to develop cosmetics. In addition, in the initial stage of acne lesions and in the stage of comedone formation, cosmetics were intended to be composed of materials that could maximize the peeling effect on dead skin cells and foreign substances on the skin.

The present invention includes a multilamellar liposome formed by mixing material A and material C and collecting them in an aqueous phase, and including material B in an oily phase, wherein the material A is an ethanol extract of the leaves of a chinensis, and the material B is Heechum's It is a carbon dioxide supercritical fluid extract, and the material C is a water extract obtained by re-extracting the extract foil generated during extraction of Heechum's carbon dioxide supercritical fluid with water, thereby providing a cosmetic composition for improving acne.

In the cosmetic composition for improving acne of the present invention, the material A and material C are preferably mixed with each other to form material D, and it is preferable to collect this in the aqueous phase.

In the cosmetic composition for improving acne of the present invention, the A or material C is preferably dissolved in each extract in butylene glycol or propanediol and collected in the water phase. good.

In the cosmetic composition for improving acne of the present invention, the material C is preferably extracted while purging nitrogen when re-extracting.

The present invention can expect an improvement effect on acne lesions by properly managing Propionibacterium Acnes known as an acne-producing bacteria. In addition, in the initial stage of acne lesions and comedone formation stage, it is possible to maximize the peeling effect on dead skin cells and foreign substances of the skin.

Figure 1 shows the process of preparing the extract used in the present invention and entrapped in liposomes.
Figure 2 shows the leaf extraction process and the huicheom extraction process in more detail.
Figure 3 shows the liposome preparation process according to the banghambeop used in the present invention.
Figure 4 is the result of measuring the polyphenol content of the chinensis leaf extract for each ethanol concentration.
5 is a result of measuring the flavonoid content of the banana leaf extract for each ethanol concentration.
Figure 6 shows the DPPH scavenging ability of Zandae extract and Heechum extract.
7 is a TEM photograph of a general liposome formulation (left) and a multilamellar liposome formulation prepared in the present invention (right).
Figure 8 is the result of measuring the cytotoxicity of the extract and huicheom extract capture liposomes.
Figure 9 is the result of measuring the collagen content (%) secreted by HS68 cells when the liposome formulations were collected from the extracts of chinensis and Heechum extract.
FIG. 10 is a result of measuring the content (%) of MMP-1 secreted by HS68 cells when the liposome formulations containing the extracts of Zanja and Heechum were treated.
Figure 11 is the anti- Staphylococcus aureus test results of the liposome formulations capturing the extracts of Zandae and Heechum extracts.
Figure 12 is the anti-Propionibacterium acnes ( Propionibacterium Acnes ) test results of the liposome formulations capturing Zandae extract and Heechum extract.
13 is a panel test result of a finished product containing liposomes encapsulated with Zandae extract and Heechum extract.

The present inventor analyzed the antibacterial efficacy of 365 kinds of herbal medicines to discover materials for improving acne.

As a result, the combination of Heechum and small bamboo leaves was finally selected, and it was confirmed that the antibacterial action against P. acnes known as the cause of acne specifically increased when used in combination rather than when they were used as individual materials. there was.

However, the antibacterial component of Heechum is evenly distributed in fat-soluble and water-soluble components, so the active ingredient cannot be completely used with a general extraction method, and it was also confirmed that a new extract manufacturing method was needed, and it was possible to develop and complete it in the present invention. there was.

Based on this, in the present invention, it was possible to develop a cosmetic intermediate material that maximizes skin permeability by trapping water-soluble extracts of fenugreek leaves and water-soluble and fat-soluble extracts of Heechum on the inside and outside of the liposome formulation.

Zandae is a perennial herbaceous plant belonging to the Bellflower family, and is also called Takju, and is called Sasam (沙蔘) in Chinese characters. Its scientific name is Adenophora triphilla var. japonica HARA. to be. It is a plant that commonly grows in the wild and is 40 to 120 cm tall, with thick roots and hairs all over. It occurs in all parts of Korea, and is also distributed in Japan, Manchuria, and China. The sprouts are eaten as herbs in the spring, and the roots are collected in early spring or autumn to be eaten roasted or used as medicine. It contains a type of saponin as an active ingredient. In the present invention, the leaves of the palm tree are used.

Siegesbeckiae Herba is an annual herb belonging to Compositae, and is the aerial part of Siegebeckia pubescens Makino, Siegebeckia glabrescens Makino and related plants of the same genus. Hicheom is known to have pharmacological effects such as anti-inflammatory action, blood pressure lowering action, anti-inflammatory action and anti-cancer action, and has already been used for the treatment of joint pain, muscle pain, and high blood pressure in oriental medicine. The major components known so far are known as diterpenes such as darutin, alkaloid, and kyrenol.

The outline of the present invention is briefly modeled and shown in Figure 1, and Figure 2 shows the leaf extraction process and the extraction process in more detail. Figure 1 shows the process of preparing the extract used in the present invention and entrapped in liposomes. Figure 2 shows the leaf extraction process and the huicheom extraction process in more detail. As can be seen in Figures 1 and 2, the palm leaf extract is an extract powder (material A) obtained by extracting the palm leaf with preferably 70% alcohol and freeze-drying it. The 70% alcohol selected as the solvent in the above was selected as the most preferable one through the measurement of the polyphenol and flavonoid contents of the zest extract.

Heechum first obtains the fat-soluble component (material B) by using the CO2 supercritical fluid extraction method to selectively obtain the fat-soluble component. This is because, as a result of comparing the content analysis of the kyrenol component for each extraction process of the Heechum extract, it was confirmed that the kylenol extraction yield of the supercritical extract was the best. In addition, the CO2 supercritical fluid extract foil is extracted using purified water and then lyophilized to obtain an extract powder (material C). At this time, it is preferable to extract while purging nitrogen. Extraction with nitrogen purge means extraction with nitrogen purge (bubble injection) into the extractor.

Material A and material C obtained above are dissolved in butylene glycol or propanediol to obtain material D. Thereafter, by applying the bangham method, material D is collected in the water phase and material B in the oil phase to form a multi-layered liposome formulation. [0033] To describe the preparation of the liposome formulation in detail, the development of the liposome formulation is based on the Bangham method, which can be modified and prepared by the following method (refer to FIG. 3). Figure 3 shows the liposome manufacturing process according to the bangham method.

Step 1: Dissolve material B, phospholipid in propanediol.

Step 2: Dry with a vacuum concentrator to form a lipid film.

Step 3: Hydrate the lipid film with an aqueous solution containing Material D.

Step 4: Use a homogenizer or ultrasonic homogenizer to homogenize the hydrated liposomal solution.

The liposome formulation of the present invention developed through the above process has excellent stability, and it was possible to secure the efficacy and stability of the collected Zanja extract and Heechum extract.

As a result of confirming the cytotoxicity of the liposomes containing the extract of Zanja chinensis and the extract of Heechum, it was confirmed that the effect of toxicity was insignificant up to 1%, confirming the degree of safety.

In addition, the collagen production and MMP-1 inhibitory efficacy of the liposomes of the extract of Zanja extract and Heechum extract were measured. .

In addition, as a result of the antibacterial activity test of the liposome extract that collected the Zanja extract and the Heechum extract, the Zanbanae extract and the Heechum extract were tested against two strains, Staphylococcus aureus and Propionibacterium Acnes . It was confirmed that both the liposome and the toner containing the liposome had antibacterial activity.

In addition, as a result of a panel test of the finished product containing the liposomes containing the extracts of Zanja chinensis and Heechum extract, it was confirmed that the product had a good feeling of use and was effective against acne.

Hereinafter, the contents of the present invention will be described in more detail through the following examples and experimental examples. However, the scope of the present invention is not limited only to the following examples and experimental examples, and includes modifications of equivalent technical ideas.

[Example 1: Establishment of Optimal Extraction Process of Zanja Leaf Extract]

(1) Sample preparation

Solvents prepared by concentrations of 0%, 50%, and 70% of ethanol were added to the raw material of the leaves of the palm tree in 10-fold ratios, extracted at room temperature for 3 hours, and the extract was centrifuged at 3,500 rpm for 10 minutes, and the supernatant was filtered through a syringe filter (0.45 um). ) was filtered.

(2) Measurement of total polyphenol and flavonoid content

① For total phenol content, 10 μL of Folin-Ciocalteau phenol reagent and 100 μL of distilled water were added to 5 μL of the extract according to the Folin-Denis method and mixed at room temperature for 3 minutes. 100 μL of 20% Na2CO3 solution was added to the mixed solution, and then reacted at room temperature for 1 hour, and the absorbance was measured at 725 nm using a spectrophotometer. Gallic acid was prepared in the concentration range of 25 ~ 800 μg / mL and analyzed in the same way as the sample, and the total phenol content of the sample extract was calculated from the standard calibration curve obtained.

② For total flavonoids, 75 μL of ethanol, 140 μL of distilled water, 5 μL of 1M potassium acetate, and 5 μL of 10% aluminum nitrate were sequentially added to 25 μL of the extract according to the method of Moreno et al. measured. The total flavonoid content of the extract was calculated from a standard calibration curve obtained in the concentration range of 1 to 500 μg/mL using Quercetin (Sigma Co., USA) as a standard material.

(3) Experimental results

Experimental results were shown in FIGS. 4 and 5 . 4 is a result of measuring the polyphenol content of the extract for each ethanol concentration, and FIG. 5 is a result of measuring the flavonoid content of the extract for each ethanol concentration.

When the ethanol concentration of the extraction solvent was 70%, it was confirmed that the content of polyphenols and flavonoids increased. Based on this, the extraction solvent was determined to be 70% ethanol, and extraction was performed.

[Example 2: Establishment of Heechum's Optimal Extraction Process]

(1) Experiment outline

The main known ingredients of Heechum include alkaloids such as darutin, esters such as kyrenol, 17-hydroxy-16alpha(-)-kauran-19-carboxylic acid, and diterpenes. there is. Among them, Kirenol is a diterpenoid found mainly in huicheom, and has anti-inflammatory, analgesic, and antibacterial effects, and is actually reported to have arthritis treatment effects and anti-obesity effects. Therefore, analysis was performed by HPLC to measure the content of kyrenol extracted from the kyrenol standard material and the Heechum extract.

(2) sample preparation

① Manufacture of Heechum supercritical extract

For supercritical fluid extraction, 100 g of Heechum was extracted for 2 hours using a CO2 supercritical fluid solvent under 400 bar and 50 °C conditions. The oil-soluble component was extracted by removing CO2 fluid as a solvent from the extract at atmospheric pressure.

② Manufacture of Heechum hot water extract

100 g of Heechum was added with 10-fold distilled water, heated to 90 ° C, and extracted with stirring for 4 hours. After centrifuging the extract at 3500 rpm for 10 min, the supernatant was filtered through a final 0.45 um filter to prepare an extract.

③ Heechum nitrogen purge extract manufacturing

After supercritical extraction of Heechum, 100 g of distilled water was added to 100 g of the remaining extract, nitrogen gas was injected and sealed to block contact with oxygen, and extraction was performed with stirring at room temperature for 4 hours. After centrifuging the extract at 3500 rpm for 10 min, the supernatant was filtered through a final 0.45 um filter to prepare an extract.

(3) Experiment method

The contents of kyrenol in Heechum supercritical extract and nitrogen purge extract were analyzed using high performance liquid chromatography (HPLC). HPLC analysis conditions were separated at a flow rate of 1 mL/min under gradient conditions using mobile phase A: 0.2% phosphoric acid in water, B: 0.2% phosphoric acid in acetonitrile. A Luna C18 (250 x 4.6 mm, 5 μm) column was used, and the wavelength used for kyrenol detection was 215 nm. As a standard material, kyrenol was prepared at a concentration of 20 to 200 μg/mL and used to prepare a calibration curve.

(4) Experimental results

As a result of analyzing the kylenol content of the Heechum hot water extract, supercritical extract and nitrogen purge extract using HPLC, no peak was found at the same retention time in the hot water extract, 96.31μg/mL for the supercritical extract and nitrogen purge extract. A concentration of 44.47 μg/mL was confirmed. Finally, it was confirmed that the content of kyrenol compared to the extract was 1.93% for the supercritical extract, which was extracted with the best yield, and 0.89% for the nitrogen purge extract.

Results of kyrenol analysis by extract extract concentration Peak Area Kirenol (ug/ml) Extraction ratio of Kirenol to extract Heechum hot water extract 5 mg/ml 0 0 0% Heechum Nitrogen Purge Extract 5 mg/ml 497.6923 44.47 0.89% Heechum Supercritical Extract 5 mg/ml 1077.9053 96.31 1.93%

From the above results, it was decided to use Heechum supercritical extract and Heechum nitrogen purge extract.

[Experimental Example 1: Determination of Antioxidant Efficacy of Fenugreek Leaf Extract and Heechum Extract]

(1) Experiment method

2,2-Diphenyl-1-picrylhydrazyl radical (DPPH) is used to confirm the antioxidant efficacy of the sample against free radicals. The electron donating ability was measured by modifying a known method. 0.2 ml of the sample was added to 0.8 ml of a 0.4 mM DPPH solution dissolved in ethanol, shaken with a vortex mixer for 5 seconds, left in the dark for 10 minutes, and then absorbance was measured at 517 nm. The electron donating ability showed the DPPH free radical scavenging ability by the following formula.

[Equation 1]

(2) Experimental results

Antioxidant efficacy of Zanja leaf extract and Heechum extract was measured by DPPH scavenging ability. Zandae extract confirmed DPPH scavenging ability of 75.98%, the highest in 70% ethanol extract, and Heechum extract confirmed DPPH scavenging ability of 82.23% and 72.44% respectively in supercritical extract and nitrogen purge extract (FIG. 6). Figure 6 shows the DPPH scavenging ability of Zandae extract and Heechum extract.

[Example 3: Development of an optimal extraction formulation capable of securing the stability of Zanja leaf extract and Heechum extract]

Each of the obtained extracts was dissolved in butylene glycol or propanediol to establish a final extract formulation with excellent stability.

[Example 4: Development of Liposome Formulation Containing Zanja Leaf Extract and Heechum Extract]

(1) Liposome formulation test according to the type of phospholipid

Among the 6 types of phospholipids (Lecico F-580 IPM, Lecico P900 IPM, Lecico Sun FM-580, Lipamin H100, Emulmetik 950, Emulmetik 930), as a result of visual observation, Emulmetik 950 formed bubbles, but did not precipitate or separate layers. , the formulation was the most stable (Table 2).

Evaluation of liposome formulations according to 6 types of phospholipids Phospholipid color formulation bubble Precipitation floor separation stability F 580 opacity Liquid X X X P900 opacity Liquid O O X Sun FM 580 opacity Liquid X O X H 100 opacity Liquid O O X EM 950 opacity Liquid O X X 1st place EM 930 opacity Liquid O X X 2nd place

(2) Preparation of liposomes

Of the six phospholipids, Emulmetik 950 was finally used as a phospholipid to prepare a liposome formulation by applying the bangham method. An oil phase extract of Heechum was added to the phospholipid phase, and liposomes were prepared by adding water-soluble Zanja leaf extract and water-soluble Heechum extract to the water phase at a concentration of 100 μg/ml, respectively.

Phospholipids used in the preparation of liposomes were lecithin obtained by hydrogenating soybean-derived lipids to remove unsaturated components. After mixing phospholipid and sodium deoxycholate in a ratio of 9:1, equal amounts of ethanol and fat-soluble extract (extract from Heechum's oil phase) were added and completely dissolved in a thermostat at 60 °C to make a transparent sol solution. After solidifying it at room temperature, it was taken to a thermostat at 60° C., and when a transparent sol solution was obtained, distilled water was added thereto and magnetic stirring was performed for 10 min or longer. It was completely dried to form a lipid film inside the flask under reduced pressure conditions at 50° C. with a vacuum concentrator. Liposomes with a final phospholipid content of 2% were completed by adding water-soluble extracts (water-soluble chinensis leaf extract, water-soluble Heechum extract) and distilled water to the lipid film to hydrate and then homogenize at 2000 rpm for 10 min.

Through this, it was possible to establish optimal liposome formulation conditions with high stability since the liposomes of multilamellar structure of aqueous phase (black) and oily phase (white) were composed of several layers (FIG. 7). 7 is a TEM photograph of a general liposome formulation (left) and a multilamellar liposome formulation prepared in the present invention (right).

[Experimental Example 2: Cytotoxicity of liposomes capturing Zandae extract and Heechum extract]

(1) Experiment method

① HS68 cell culture

HS68 cells, which are human-derived skin fibroblasts, were purchased from Korea Cell Line Bank and used. HS68 cells were cultured in a DMEM medium (Welgene) in a 37°C humidified CO2 incubator (5 % CO2/95% air). When the cells were about 80% full of the culture dish, the cell monolayer was washed with phosphate buffer saline (PBS, pH7.4), and trypsin-2.65 mM EDTA was added to detach and subculture the cells, and the medium was exchanged every 2 days. did

② Measurement of cell viability of HS68 cells

HS68 cells were dispensed in a 48-well plate at 1 × 104 cells/well, and the cells were cultured for 24 hours. After culturing the cells for 24 hours, the cells were cultured for 24 hours by replacing them with serum-free cell culture medium containing various concentrations of each test substance. After culturing the cells for 24 hours, MTT assay (Denizot F and Lang R. J Immunological Method 89:271-277, 1986) was performed to measure the number of viable cells. The MTT assay method is based on the principle that mitochondrial dehydrogenase reduces MTT (Amresco) to produce blue-colored formazan. In this test, formazan was dissolved in isopropanol and absorbance was measured at a wavelength of 570 nm.

(2) Experimental results

In order to investigate the cytotoxicity of liposomes in HS68 cells, the liposome formulations were treated with cell culture medium at various concentrations and cultured for 24 hours, followed by MTT assay. Cell viability was not affected when the liposome formulation was treated at a concentration of 0.1 to 1%. When the 2% liposome formulation was treated, it was confirmed that the cell viability significantly decreased to 67.6%. Figure 8 is the result of measuring the cytotoxicity of the liposomes capturing the extract and Heechum extract.

[Experimental Example 3: Collagen production and MMP-1 inhibitory efficacy of liposomes capturing Zanja extract and Heechum extract]

(1) Experiment method

HS68 cells were dispensed in a 24-well plate at 1 × 105 cells/well, and the cells were cultured for 24 hours. After culturing the cells for 24 hours, each acerola extract, Manuka oil, and liposome formulation were exchanged for serum-free cell culture medium containing various concentrations, and the cells were cultured for 24 hours. After culturing the cells for 24 hours, the cell culture medium was collected and the collagen type I C-Peptide (PIP) EIA Kit (Takara) and Human MMP-1 ELISA Kit (Sigma-Aldrich) were used, respectively, according to the method suggested by the manufacturer. and MMP-1 content were measured.

(2) Experimental results

① Collagen is a major protein distributed in all connective tissues and is abundant in skin, bone, and ligaments. In order to investigate the effect of the liposome formulation on the collagen production and secretion of HS68 cells, which are skin fibroblasts, the cells were cultured by treating with various concentrations of the test substance, and then the amount of collagen secreted by the cells was measured. It was confirmed that the collagen content of the liposome formulation of the present invention increased in a concentration-dependent manner. Figure 9 is the result of measuring the collagen content (%) secreted by HS68 cells during the treatment of the liposome formulations capturing the extracts of chinensis and Heechum extracts.

② MMP-1 is a collagen degrading enzyme. In order to investigate the effect of liposome formulations on the production and secretion of MMP-1 by HS68 cells, which are skin fibroblasts, the cells were cultured by treating them with various concentrations, and then the cells secreted MMP. -1 The content of protein was measured. When the liposome formulations of the present invention were treated at concentrations of 100, 500, and 1000 μg/mL, the content of MMP-1 produced and secreted from HS68 cells tended to decrease in a concentration-dependent manner. FIG. 10 is a result of measuring the content (%) of MMP-1 secreted by HS68 cells when the liposome formulations were treated with the extract of Zanjata and Heechum extract.

[Experimental Example 4: Antibacterial activity (anti-acne bacteria inhibitory activity) test of liposome extracts containing Zandae extract and Heechum extract]

(1) Experiment outline

An experiment was conducted to determine whether the liposomal formulation of Zandae and Heechum extracts had antibacterial activity, and to determine whether antibacterial activity was maintained even when applied to toner. An antibacterial test was conducted using two aerobic bacteria, Staphylococcus aureus and acne-causing strain, Propionibacterium Acnes . A DISC experiment was conducted as a method for measuring the antibacterial activity of the extract.

(2) Experiment method

① After steaking P. acnes on RC agar medium and S. aureus on TSB agar medium, they were incubated at 37℃ for 24 h. Since P. acnes is an anaerobic bacterium, it was cultured under anaerobic conditions.

② SPL Round-Bottom polystyrene 5ml tubes were filled with 2ml of sterilized 0.85% NaCl solution in one tube and 1ml in the other three tubes.

③ After inoculating about 10 colonies of each grown bacteria in NaCl solution, vortexing and measuring the absorbance at 630 nm to adjust the concentration of the bacterial solution.

④ Streak evenly 8 times with Muller-Hinton agar (dispensed in 17 ml per 90 mm plate, height 4 mm) rotating 90° each. At this time, streaking was performed after removing as much moisture as possible from the cotton swab.

⑤ 4 sterilized DISCs were placed at regular intervals and 20 μl of each sample was inoculated into the DISCs (the samples were prepared by dissolving them in 50% ethanol solvent in a freeze-dried state at a concentration of 100 μg/ml).

⑥ After confirming that the disc was well placed, it was incubated at 37℃ for 24 hours.

⑦ The diameter of the area inhibiting bacterial growth was measured for each sample.

sample number sample Etc One Remnant, Heechum hot water extract (100㎍/1ml) control group 2 Zandae extract, Heechum extract collection liposome (100㎍/1ml) Experiment group 1 3 toner control group 4 Toner (including liposomes containing Zandae extract and Heechum extract) experimental group 2

(3) Sample preparation

① Zandae, Heechum hot water extract

Zandae and Heechum were weighed with 10-fold purified water, extracted at 90 ° C for 4 hours, and then filtered using a 0.45 μm filter. The extract was obtained as a powder after lyophilization. The obtained powder was prepared by dissolving it in a 50% ethanol solvent at a concentration of 100 ug/ml.

② Zandae extract and Heechum extract collection liposome

It was prepared by dissolving the lyophilized extract and liposomes of Heechum extract prepared in Example 4 in a 50% ethanol solvent at a concentration of 100 μg/ml in a lyophilized powder state.

③ Toner (including liposomes that capture Zandae extract and Heechum extract)

It was prepared by dissolving the Zandae extract and Heechum extract trapping liposomes prepared in Example 4 in a toner developed by the company at a concentration of 100 ug/ml.

(4) Experimental results

Staphylococcus aureus ( Staphylococcus aureus ) Experimental results are shown in FIG. 11 . Propionibacterium acnes ( Propionibacterium Acnes ) Experimental results were shown in FIG. 12 . As a result of the experiment, it was confirmed that the liposomes in which the extracts of Zandae and Heechum were collected had antibacterial activity, and it was confirmed that the antibacterial activity was maintained without losing even when applied to toner.

[Example 5: Preparation of a finished product containing liposomes encapsulated with Zandae extract and Heechum extract ]

The Dr. The March Manuka Spot Gel, to which liposomes containing Zandae extract and Heechum extract were added, was manufactured as a prototype through the following process. Weigh phase A in a beaker, heat it at 70 ° C for about 30 minutes to completely dissolve it, and when cooled to 50 ° C, add phase B and mix with an azimixer. When the gel formulation is cooled to room temperature, phase C is added and mixed with an azimixer for 10 minutes. As a result of the usability test, sample 1 had too strong viscosity and sample 2 had too weak viscosity, so sample 3 had moderate hardness and was applied smoothly.

raw material name content(%) A
Purified water 80.87 81.07 80.97 carbomer 0.25 0.15 0.20 sodium carbomer 0.25 0.15 0.20 allantoin 0.10 0.10 0.10 betaine 0.50 0.50 0.50 Niacinamide 2.00 2.00 2.00 glycerin 3.00 2.50 2.00 Butylene Glycol 3.00 3.50 4.00 NS-3000 1.00 1.00 1.00 D-panthenol 0.50 0.50 0.50 hyaluronic acid 0.10 0.10 0.10 Carvasol 0.50 0.50 0.50 EDTA 0.05 0.05 0.05 B HCO-40 0.50 0.50 0.50 1,2-Hexanediol 1.00 1.00 1.00 Ethylhexylglycerin 0.03 0.03 0.03 tea tree oil 0.30 0.30 0.30 time oil 0.05 0.05 0.05 manuka oil 1.00 1.00 1.00 ethanol 4.00 4.00 4.00 C Liposome raw material in which the Zandae extract and Heechum extract of the present invention are captured 1.00 1.00 1.00 Sum 100.00 100.00 100.00 sample 1 sample 2 sample 3

[Experimental Example 5: Panel Test of Finished Product Containing Liposomes Encapsulating Zandae Extract and Heechum Extract]

The Dr. The March Manuka Spot Gel prepared in Example 5 was recruited by a group of 20-year-olds suffering from acne, spread evenly on the troubled area for about 2 weeks and absorbed, collecting opinions on before and after photos and overall reviews. The results were as shown in FIG. 13 .

Claims (4)

Including multilamellar liposomes formed by mixing material A and material C and collecting them in an aqueous phase, and including material B in an oil phase,
The material A is an ethanol extract of the leaves of the palm tree,
The material B is Heechum's carbon dioxide supercritical fluid extract,
The material C is a cosmetic composition for improving acne, characterized in that it is a water extract obtained by re-extracting the extract foil generated during extraction of Heechum's carbon dioxide supercritical fluid with water.
According to claim 1,
The material A and material C,
A cosmetic composition for improving acne, which is characterized in that it is mixed with each other to produce material D and collected in the water phase.
According to claim 1,
The above A or material C,
A cosmetic composition for improving acne, characterized in that each extract is dissolved in butylene glycol or propanediol and collected in an aqueous layer.
According to claim 1,
The material C,
Upon re-extraction, a cosmetic composition for improving acne, characterized in that extracted while purging nitrogen.

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