KR101678152B1 - Antimicrobial preservatives for cosmetic containing lipid vesicle-encapsulated fatty acid tripeptide salt and method for manufacturing the same - Google Patents

Abstract

The present invention relates to an antimicrobial preservative for cosmetics comprising a fatty acid tripeptide salt encapsulated in a lipid vesicle and a method for preparing the antimicrobial preservative, and more particularly, to an antimicrobial preservative for a cosmetic comprising encapsulating a fatty acid tripeptide salt having excellent antibacterial activity in a lipid vesicle, The present invention relates to an antimicrobial preservative for cosmetics and a method for producing the same.
According to the present invention, the antimicrobial activity is excellent, but when added to cosmetics, the fatty acid tripeptide salt such as palmitoyl tripeptide, which is difficult to use as a cosmetic preservative due to the formation of a precipitate by reaction with the thickening agent, Thereby solving the problem of forming a precipitate and recovering the reduced antibacterial activity. Accordingly, a fatty acid tripeptide salt such as palmitoyl tripeptide or the like can be used as an antimicrobial preservative for cosmetics.

Description

FIELD OF THE INVENTION [0001] The present invention relates to an antimicrobial preservative for cosmetics comprising a fatty acid tripeptide salt encapsulated in a lipid bilac, and an antimicrobial preservative for a cosmetic comprising the fatty acid tripeptide salt encapsulated in a lipid vesicle,

The present invention relates to an antimicrobial preservative for cosmetics comprising a fatty acid tripeptide salt encapsulated in a lipid vesicle and a method for preparing the antimicrobial preservative, and more particularly, to an antimicrobial preservative for a cosmetic comprising encapsulating a fatty acid tripeptide salt having excellent antibacterial activity in a lipid vesicle, The present invention relates to an antimicrobial preservative for cosmetics and a method for producing the same.

Cosmetics contain various organic nutrients that are nutrients of microorganisms in addition to water. Since they come into contact with air and human hands while closing the lid every time they are used for a long period of time, microorganisms such as bacteria and fungi multiply, . When cosmetics are contaminated with microorganisms, it is necessary to add a preservative to protect the cosmetics from deterioration, deodorization and the like caused by microorganisms, since the quality abnormality such as odor, color, texture, viscosity change, and fungus occur. Preservatives are used for the purpose of preventing the degradation of products by inhibiting the growth of microorganisms introduced from the outside, but they can cause skin irritation, and they are basically cytotoxic. Therefore, they are harmful to the human body It is necessary to mix the product in a minimum effective amount in consideration of the safety and stability of the product.

Paraoxybenzoate, a typical preservative used in cosmetics, is effective against various bacteria and fungi, has no odor, and does not cause discoloration, hardening, turbidity, and is relatively low in toxicity. It is also widely used in cosmetics because of its advantages such as low cost, chemical stability at a wide pH range and high temperature, and effective use of methylparaben, ethylparaben, butylparaben, etc. come. However, the safety of parabens has been continuing because the parabens can cause allergic reactions or hormonal changes as well as accumulate in the body and cause cancer in the long term. Recent studies have shown that parabens have a similar structure to that of estrogen, a female hormone, which may disturb the endocrine system, and men may have abnormalities in reproductive function, such as decreased sperm count, It has also been reported that it can cause side effects such as causing contact allergy to patients with It has also been reported that parabens are absorbed into the human body, which may be a potential cause of breast cancer in women, and that parabens are commonly detected in tumor cells of actual breast cancer patients.

Isopropyl alcohols used as a substitute for parabens preservatives may also irritate mucous membranes and cause skin allergies. In addition, various side effects have been reported on existing preservatives such as imidazolidinyl urea, diazolidinyl urea, and phenoxyethanol. Despite these side effects, there is a growing interest in the antimicrobial activity of peptides as an alternative preservative that is relatively safe and less irritating, since no preservatives can be incorporated due to safety concerns associated with long-term use of cosmetics. Antimicrobial peptides have been found to have a wide range of applications, are environmentally friendly, have no residual toxicity, and have no resistance to pathogens. The poly cationic peptide compound showed antimicrobial activity against prokaryotic and eukaryotic microorganisms, and the peptide derived from the growth factor had antimicrobial effect on not only fungi but also gram negative and positive bacteria. Although melittin isolated from bee venom has excellent antimicrobial activity, it is toxic to various cells including human erythrocytes. Therefore, there are many limitations in application of this antimicrobial peptide as a preservative.

[Document 1] O. Gortzi, S. Lalas, I. Chinou, and J. Tsaknis, Evaluation of the antimicrobial and antioxidant activities of Origanum dictamnus extracts before and after encapsulation in liposomes, Molecules, 12, 932-945 (2007). [2] P. S. Malheiros, V. Sant'Anna, M. Barbosa, A. Brandeli, and B. D. G. Melo Franco, Effect of liposome-encapsulated nisin and bacteriocin-like substance P34 on Listeria monocytogenes growth in Minas frescal cheese, Int. J. Food Microbiol., 156, 272-277 (2012)

The present invention provides an antimicrobial preservative for cosmetics comprising an encapsulated fatty acid tripeptide salt, wherein the fatty acid tripeptide salt having excellent antibacterial activity is encapsulated in a lipid vesicle to remarkably improve the antibacterial activity in cosmetic formulations, and a method for producing the antimicrobial preservative.

The present invention provides an antimicrobial preservative for cosmetics comprising a fatty acid tripeptide salt encapsulated in a lipid vesicle.

The fatty acid tripeptide salt may be palmitoyl tripeptide (trade name M330).

The antimicrobial preservative for cosmetics comprises a fatty acid tripeptide salt in a concentration of 1 to 4% (w / w), and the fatty acid tripeptide salt is encapsulated in a lipid bicycle. When the concentration of the fatty acid tripeptide salt is encapsulated in the above-described range, no precipitation occurs upon mixing with the cosmetic, and the antibacterial activity is increased.

The lipid bilayer membrane is composed mainly of a phospholipid dissolved in a small amount of ethanol and contains cholesterol together with the phospholipid. The cholesterol may be included in a ratio of 5 to 20% (w / w) based on the total weight of lipids. More preferably about 10% (w / w) or more. A stable lipid vesicle may be formed if the above range is maintained.

The lipid vesicle comprising the fatty acid tripeptide salt may further comprise a chelating agent.

The chelating agent may include one or more selected from the group consisting of EDTA, sodium polyphosphate, sodium metaphosphate, citric acid and gluconic acid.

The chelating agent may be contained in an amount of 1/10 to 1/2 of the weight of the fatty acid tripeptide salt.

The lipid vesicle may be an etosome.

The present invention relates to a process for producing a solution (step a) in which a mixture of a phospholipid and a cholesterol is dissolved in ethanol; Adding an aqueous solution in which the fatty acid tripeptide salt is dissolved to the solution of step (a) to form a hydrated liquid crystalline phase and dispersing it to form a lipid vesicle particle in which the fatty acid tripeptide salt is captured (step b And a method for producing the antimicrobial preservative for cosmetics.

In step (a), a mixture of phospholipid and cholesterol is dissolved in ethanol at a high temperature (55-70 ° C), and an aqueous solution in which a fatty acid tripeptide salt is dissolved is added to form a hydrated liquid crystalline phase Can be performed.

The step (b) may be carried out by dispersing the liquid crystal phase of step (a) in water to form a lipid vesicle particle in which the fatty acid tripeptide salt is captured.

The phospholipids and cholesterol may be mixed at a weight ratio of 9.5 to 8.0: 0.5 to 2.0 and the mixture may be prepared by dissolving the mixture at a weight ratio of 1: 1 in ethanol.

The fatty acid tripeptide salt may be a palmitoyl tripeptide.

The aqueous solution in which the fatty acid tripeptide salt is dissolved may further contain a chelating agent. When a chelating agent is additionally contained, the step (a) is a step of dissolving a mixture of phospholipid and cholesterol in ethanol at a high temperature (55-70 ° C), adding an aqueous solution in which a fatty acid tripeptide salt and a chelating agent are dissolved, lt; RTI ID = 0.0 > hydrated < / RTI > liquid crystalline phase.

The chelating agent may include one or more selected from the group consisting of EDTA, sodium polyphosphate, sodium metaphosphate, citric acid and gluconic acid.

The antimicrobial preservative for cosmetics may be contained in an amount of 2 to 5% (w / w) based on the total weight of the cosmetic composition. If the amount is less than the above range, the antimicrobial preservative effect may not be sufficiently exhibited, and if it is more than the above range, it may be difficult to maintain the cosmetic formulations.

According to the present invention, the antimicrobial activity is excellent, but when added to cosmetics, the fatty acid tripeptide salt such as palmitoyl tripeptide, which is difficult to use as a cosmetic preservative due to the formation of a precipitate by reaction with the thickening agent, Thereby solving the problem of forming a precipitate and recovering the reduced antibacterial activity. Accordingly, a fatty acid tripeptide salt such as palmitoyl tripeptide or the like can be used as an antimicrobial preservative for cosmetics.

FIG. 1 is a graph comparing antimicrobial effects of a peptide preservative, M330 (palmitoyl tripeptide), methylparaben, and phenoxyethanol.
Fig. 2 is a photograph showing the appearance of (a) before and after (b) the addition of M330 to the cosmetic formulation of the essence formulation.
FIG. 3 is a graph comparing the decrease in viscosity when the M330 solution is added to the makeup of the essence formulation and the decrease in viscosity when the same amount of distilled water is mixed instead of the M330 solution.
FIG. 4 is a graph showing the particle size of lipid vesicles (Examples 1 to 8) according to the concentration of M330 (palmitoyl tripeptide) and the HPC / cholesterol mixing ratio measured by a particle size analyzer (PSS, Nicomp 380).
5 is a photograph showing the appearance of lipid bilayer solution (Example 1) in which M330 was collected at a concentration of 2% and added to cosmetic formulations of the essence formulation.
FIG. 6 is a graph (M330 2%) comparing changes in viscosity of a cosmetic formulation of the essence formulation (M330) mixed with lipid vesicle encapsulation (Example 1) and without encapsulation (Comparative Example 1).
7 is a graph showing the results of comparing the antibacterial activity of M330 (Example 2 and Comparative Example 2) against various microorganisms in a lotion-form cosmetic. (a) E. coli, (b) P. aeruginosa , (c) S. aureus, and (d) C. albicans .
8 is a graph showing the results of measuring particle sizes of lipid vesicles (Examples 9 to 14) according to the ratio of M330 / EDTA and HPC / Cholesterol.
Figure 9 compares the antimicrobial activity of non-encapsulated M330 / EDTA (Comparative Example 3) and encapsulated M330 / EDTA (Example 9) ( C. albicans ).
10 is a graph showing the antibacterial effect of M330 / EDTA (Examples 9 to 11 and Comparative Examples 3 to 5) according to the addition ratio of EDTA. (a) 6/1, (b) 3/1, (c) 2/1.

Hereinafter, the present invention will be described in more detail with reference to Examples. The objects, features and advantages of the present invention will be readily understood through the following examples. The present invention is not limited to the embodiments described herein, but may be embodied in other forms. The embodiments described herein are provided to enable those skilled in the art to fully understand the spirit of the present invention. Therefore, the present invention should not be limited by the following examples.

Examples and Comparative Examples

Sample Preparation

A method for synthesizing the palmitoyl tripeptide (M330) antimicrobial peptide used in the present invention as a product synthesized and developed by Miwon Company is briefly described as follows. For the solid-phase reaction, the first amino acid (lysine) protected at the N-terminus with 9-fluorenylmethyloxycarbonyl (Fmoc) and the side chain with butyloxycarbonyl (Boc) is added to a rink amide resin with N , N'-diisopropylcarbodiimide (DIC) and 1-hydroxybenzotriazole (HOBt) as an additive. The protecting group at the N-terminus was removed using piperidine In the same way, the second and third lysine were coupled. Subsequently, the palmitic acid was coupled in the same manner and then separated from the solid resin using a cleavage solution containing trifluoroacetic acid (TFA), followed by precipitation and separation using diethyl ether M330 was obtained (Lee, Yoon-Seop and Choi, Hyeongjung, Fatty acid tripeptide salt and antimicrobial composition containing the same, Patent, 10-1151878, 2012).

Phospholipid was Emulmetik 950 (Lucas Meyer) with 95% or more of PC (Phosphatidyl Choline) as a lecithin, which eliminated the unsaturated component by hydrolyzing lipid extracted from soybean. For convenience, the name was changed to HPC (Hydrogenated Phosphatidyl Choline). Cholesterol (Chol.) Was used in Sigma, and ethanol was used in Daejeong Chemical. In addition, oily raw materials, moisturizers, surfactants, and thickeners necessary for cosmetic production were used for cosmetics.

The fungus is C. albicans (yeast, ATCC 10231), a Gram-negative bacteria P. aeruginosa (Pseudomonas aeruginosa, ATCC 27853) and E. coli (E. coli, ATCC 8739) to the fungi used in the challenge test and M1C (Challenge Test), Gram positive bacteria were allotted to S. aureus (ATCC 6538p), and the microorganism resource center (KCTC). Plate count agar (PCA) and plate dextrose agar (PDA) were used for bacterial culture, and MHB (Mueller Hinton broth) was used for liquid medium. All of them were Becton, Dickinson and Compan y products.

Example: Preparation of lipid vesicle harvested M330

Various lipid vesicles were prepared as follows by varying the concentration of M330, the mixing ratio of HPC and cholesterol, as follows. The HPC / cholesterol mixture and ethanol are placed in a round bottom flask at the same weight ratio and dissolved in a constant temperature chamber at 60 ° C to form a clear solution. Add an amount of M330 (or a mixture of M330 and EDTA) equal to the amount of ethanol, and then agitate the solution for 5 minutes. Then, the solution becomes a gel state and a white hydrated liquid crystalline phase is formed . Next, when distilled water is slowly added by using a constant amount pumping pump while the agitator is operated in the dispersion step of hydrated liquid crystal phase, the hydrated liquid crystal phase is dispersed in water as vesicle-like particles, which becomes a lipid vesicle in which M330 is captured . The finished lipid vesicles are slowly cooled at room temperature while stirring continuously (see Table 1).

Comparative Example: A non- encapsulated M330, a mixture of M330 and EDTA, methylparaben was used as a comparative example (see Table 1). In the case of the comparative example, an aqueous solution was prepared without encapsulation.

Lipid
Vesicle
Encapsulation M330 (%, w / w) ¹⁾ EDTA ²⁾ HPC / Chol. rario ³⁾ Other Example 1 ㅇ 2 - 9: 1 Example 2 ㅇ 3 - 9: 1 Example 3 ㅇ 4 - 9: 1 Example 4 ㅇ 5 - 9: 1 Example 5 ㅇ 2 - 8: 2 Example 6 ㅇ 3 - 8: 2 Example 7 ㅇ 4 - 8: 2 Example 8 ㅇ 5 - 8: 2 Example 9 ㅇ 3 1/6 9: 1 Example 10 ㅇ 3 1/3 9: 1 Example 11 ㅇ 3 1/2 9: 1 Example 12 ㅇ 3 1/6 8: 2 Example 13 ㅇ 3 1/3 8: 2 Example 14 ㅇ 3 1/2 8: 2 Comparative Example 1 x 2 - - Comparative Example 2 x 3 - - Comparative Example 3 x 3 1/6 - Comparative Example 4 x 3 1/3 - Comparative Example 5 x 3 1/2 - Comparative Example 6 x - - - Methyl paraben 0.2%

1) The concentration of M330.

2) The mixing ratio of EDTA to the weight of M330.

3) The mixing ratio (weight ratio) of HPC and cholesterol.

Test Example: MIC and Challenge Test

Determination of the minimum inhibitory concentration (MIC) was carried out by a two-fold serial dilution method. Each bacterium was inoculated into a liquid medium and incubated at a suitable temperature and for a certain period of time, and diluted to a concentration of 2 × 10 ⁶ CFU / mL. Add 50 μL of the liquid medium to all the columns of the 96-well plate. Add 50 μL of the solution containing the antimicrobial agent at 8% concentration to the first column of the 96-well plate and mix well. After mixing, 50 μL was taken from the first column, followed by mixing in the next two columns. The same procedure was repeated repeatedly, and the diluted solution was diluted 1/2 times with respect to all the columns. Then, 50 μL of the culture solution in the liquid medium Add to heat. The 96-well plate was incubated in a 37 ° C incubator for one day and the absorbance at 540 nm was measured using an ELISA reader (Synergy HT, Bio-Tek) to determine the MIC of each strain.

The challenge test was conducted with reference to KFDA's 'Microbiological Limit Standards and Test Method Guidelines'. The concentration of bacteria to be tested in cosmetic lotion formulation containing antimicrobial agent is approximately 10 ⁶ - 10 ⁷ CFU / mL To be A certain amount of the bacterial solution was added, and the mixture was well mixed and stored in an incubator at 35 ° C. 100 μL of the stored lotion was dispersed in E-tube and dispersed with 10 times of distilled water. 100 μL of the lotion was applied to a sterilized solid medium and incubated in an incubator at 35 ° C. for one day. The number of grown colonies was expressed in CFU / mL in terms of dilution factor. The buoyancy was confirmed by comparing the degree of death of the inoculum with the passage of time, and it was judged that the preservative effect was obtained when 99.9% of the first bacteria were killed within one week.

Cosmetics Manufacturing

Cosmetics were made into two types of essence and lotion. The composition of each is shown in Tables 2 and 3. The preparation of the essence was carried out as follows. The beaker was filled with glycerine, propylene glycol, sodium hyaluronate (1%), carbopol 940 (carboxy vinyl polymer, increasing 2%) and distilled water. Add silicone oil, Tween 80 (polyoxyethylene sorbitan monooleate), which is an oily substance, to the other beaker with ethanol and dissolve all at a temperature of 60 ° C. While both phases were transparent, the oil phase was slowly added to the beaker of the water-based material and stirred uniformly with an agitator to effect solubilization. Then, triethanolamine (TEA) was added thereto and stirred for 10 minutes and then slowly cooled at room temperature . Lotion formulations were prepared as follows. Add the water-based materials such as glycerin, butylene glycol, hyaluronic acid (1%), carbopol 940 (carbopol 940, 2%) and distilled water in a beaker and mix well at a temperature of 70 ° C. Oily materials such as stearic acid, olive oil, cetostearyl alcohol, cetyl ethylhexanoate (CEH), Arlacel 60 (sorbitan monostearate) and Tween 80 are also put into beakers All at a temperature of 70 ° C. When both phases are in a transparent state, the aqueous phase is mixed well with a homomixer, the emulsion is slowly added to the emulsion, and the emulsion is neutralized by adding TEA.

Ingredient contents (g) glycerine 8.5 propylene glycol 7.5 sodium hyaluronate (1%) 8.0 트리 에탄anol 0.12 carbopol 940 (2%) 6.0 ethanol 8.0 silicone oil 0.1 Tween 80 0.4 D.W. up to 100

Ingredient contents (g) glycerine 3.0 butylene glycol 2.0 sodium hyaluronate (1%) 1.0 트리 에탄anol 0.1 carbopol 940 (2%) 5.0 stearic acid 1.0 cetostearyl alcohol 1.9 cetyl ethylhexanoate 7.0 olive oil 2.0 Arlacel 60 0.7 Tween 80 2.8 D.W. up to 100

Cosmetic application and encapsulation of M330

Figure 1 compares the antimicrobial effect of the peptide preservative M330 with the antimicrobial effects of methylparaben and phenoxyethanol, which were previously used in cosmetics, and the minimum inhibitory concentrations ( MICs ) of three strains of C. albicans, P. aeruginosa , and S. aureus MIC). M330 has strong antimicrobial properties that can inhibit all three bacteria even at low concentrations compared to existing cosmetic preservatives. In particular, M330 has a minimum inhibitory concentration of several ppm required to inhibit bacteria such as P. aeruginosa and S. aureus, and is capable of suppressing bacterial proliferation even at a concentration as low as about 1000 times lower than that of methylparaben or phenoxyethanol Showed antibacterial effect. As such, M330 has excellent antimicrobial properties and was expected to be used as a cosmetic preservative. However, when applied to cosmetic formulations, various problems occurred. One of them, as shown in Fig. 2, when M330 was added to the transparent cosmetic essence formulation (a), formation of a white precipitate as apparent in (b) was confirmed. Another phenomenon that appears in formulations when M330 is added is a decrease in viscosity. As shown in Fig. 3, when the distilled water was mixed with the distilled water at a ratio of 3% (w / w), there was almost no change in viscosity, but an aqueous solution of M330 (concentration of 2% (w / w)) was mixed at a ratio of 3% Shear thinning properties of the newtonian fluid showed a slight viscosity only in a very small shear rate region and a nearly constant viscosity value. This is because the carbopol which increased the viscosity in the essence formulation lost its function as an increasing agent by the addition of M330 and precipitated.

Because carbopol is a polyacrylic acid component, the hydrogen ion is separated from the carboxylic acid group in the neutral state water and is dissolved in water with a negative charge and hydrodynamic volume is increased. The viscosity of the formulation increases. M330 is positively charged in water due to the presence of the amino group of lysine on the peptide structure. Therefore, when M330 is added to the essence, sedimentation occurs due to the electrostatic bonding between the negative charge of the in-essence carbopol and the positive charge of M330, thereby reducing the hydrodynamic volume of the carbopol, thereby greatly reducing the viscosity of the essence .

In order to prevent precipitation in cosmetics due to the addition of M330, encapsulation of a lipid liquid crystal type beacle, lipid Vicyclo M330, was attempted (see the above example). In order to make the optimal lipid vesicle, the lipid vesicle particle size changes were examined by varying the ratio of constituents. The size of the vesicle particles formed when the lipid vesicles were prepared by varying the lipid and ethanol contents of the Vesicle membrane and varying the M330 concentration and the lipid cholesterol ratio were measured by a particle size analyzer (PSS, Nicomp 380) As shown in Fig. Although not shown in the figure, when the vesicles were made solely with phospholipid (HPC) without cholesterol, the vesicles were made in an unstable state with a gel state or a particle size of several μm or more. A stable vesicle of 200 nm or less was produced regardless of the cholesterol mixing ratio of 10% (w / w) and 20% (w / w) when M330 was prepared at a concentration of 2% (w / w). However, when the cholesterol is mixed at 10% (w / w) at a concentration of 3% or more of M330, a relatively stable sized vesicle of several hundred nano size is produced, but when 20% (w / It grows rapidly. A cholesterol mixture of about 10% (w / w) helps to form a stable vesicle membrane while adding some hydrophobicity to the HPC, but when the mixing ratio is above a certain level, it forms an entangled structure with the HPC, The cholesterol mixing ratio is suitably about 10% (w / w) because it becomes an obstruction. FIG. 5 is a photograph of the lipid vesicle solution (M330 2%) in which M330 was captured and mixed with the essence. Unlike the case where M330 was directly added, no precipitation was observed when the lipid vesicle was encapsulated and added to the essence . However, if the concentration of M330 is 5% (w / w) or more, the miscibility between the constituents may be lowered and the vesicle may not be properly formed.

The viscosities were compared (M330 2%) when lipid vesicle encapsulated M330 (Example 1) and non-encapsulated M330 (Comparative Example 1) were each mixed in an essence formulation. As shown in FIG. 6, when the capsules were encapsulated and mixed with lipids, the viscosity was relatively higher than that when they were mixed without encapsulation. Encapsulating M330 limits the direct contact between the M330 and the carbopole to inhibit the precipitation formation, so that the hydrodynamic volume of the carbopol can be maintained and the viscosity values are relatively high.

Analysis of Antimicrobial Effect of M330 (Palmitoyl Tripeptide) in Cosmetic Formulation

A challenge test was conducted to compare the antibacterial activity of M330 against various microorganisms in cosmetic formulations. The cosmetics are made into lotion formulations, which are made up of 3% (w / w) M330 in lipid vesicle encapsulated form and 3% (w / w) . In this case, M330 was added at a ratio of 0.09% (w / w) based on the total weight of the cosmetics, and the antimicrobial effect against the four kinds of bacteria was examined by changing the number of viable cells through the challenge test (FIG.

Gram-negative bacteria such as E. coli In the inoculated lotion (a), 99.9% of the bacteria inoculated within 3 days were killed, and when the M330 was encapsulated in lipid vesicles, the number of bacteria decreased slightly during the reduction of the bacteria. In a lotion (b) inoculated with P. aeruginosa , 99.9% of the bacteria inoculated in one day were killed. In particular, when the M330 was encapsulated, the bacterial number was significantly reduced compared to the non-encapsulated aqueous solution And 99.9% of the bacteria inoculated in 6 hours were killed. The results of the measurement of the number of viable cells in the lotion inoculated with gram-positive S. aureus showed no effect of encapsulating M330 in (c), but most of the inoculated bacteria, regardless of encapsulation, mostly died within 4 days to 5 days. In the case of the lotion (d) inoculated with the fungus C. albicans , no bacterial decline was observed after 7 days of inoculation and no significant difference was observed in the aqueous solution of M330 or in the case of encapsulation with vesicle. For comparison, we tested the 0.2% concentration of methylparaben, which is commonly prescribed in cosmetics, and all the bacteria were killed after 6 days. These results suggest that M330 can be used in cosmetic formulations such as E. coli, P. aeruginosa, S. aureus It showed strong antimicrobial effect against bacteria but no antimicrobial effect against fungus C. albicans .

The reason why the antibacterial effect is increased by the encapsulation of M330 can be explained as follows. In cosmetic formulations, M330 loses its antibacterial activity due to its strong mutual attraction with the thickening agent. However, when M330 is encapsulated, contact with external substances is blocked, so that the antibacterial ability is prevented to some extent, , It is predicted that M330 will contribute to the increase of antibacterial activity by allowing access to bacteria and penetration into bacterial membrane. However, yeast-like fungi, C. albicans, are composed of thick, firm fibrous cell walls and, in this case, the M330 itself lacks antimicrobial activity whether encapsulated or not.

In order to supplement the deficiency of M330 against C. albicans , chelating agents were mixed with M330. In this example, EDTA was used as a chelating agent. EDTA helps to maintain the stable phase by blocking the deterioration and discoloration of cosmetics by blocking the ionic activity of metal components in cosmetic formulations, By reducing the concentration of metal salts such as Zn, it affects the growth and proliferation of bacteria. EDTA was added to M330 at a concentration of 3% (w / w) in 0.5%, 1% and 1.5%, respectively. 3, 1/2 ratio. FIG. 8 shows that when the amount of cholesterol added was 10%, the amount of vesicle particles tended to decrease as the ratio of EDTA was increased. When the amount of cholesterol added was 20% , Unstable particles such as more than 2 μm were formed irrespective of the EDTA addition ratio.

(W / w) ratio to each of the lotion formulations, which were not encapsulated but dissolved in an aqueous solution (Comparative Examples 3 to 5), were added to the lotion formulations, respectively, by viclocc encapsulating the M330 / EDTA mixture (Examples 9 to 11) The challenge test was conducted only for C. albicans . FIG. 9 is a photograph showing a comparison of the results of a challenge test conducted with a lotion inoculated with C. albicans . When M330 and EDTA were added without encapsulation, the bacterium still remained after 7 days. However, when the mixture was added with encapsulated lipid vesicle, the bacteria were completely killed and not observed. FIG. 10 shows the results of a change in the number of viable cells according to the EDTA addition ratio in the C. albicans challenge test. In the case of aqueous solution without M330 / EDTA (6: 1) encapsulation, there was a large amount of bacteria on the 7th day, but in the same concentration of Vesicle solution, 99.9% of the bacteria inoculated on the 7th day were killed and the antibacterial effect was better (A). When EDTA is added at a higher concentration ratio, most of the C. albicans bacteria die within 7 days even in the non-encapsulated aqueous solution. In the case of encapsulation, most of the bacteria are killed in 0.25% (B, c). In addition, the antimicrobial effect was more rapid than that of the control. EDTA is not recognized as a cosmetic preservative because it does not have strong antimicrobial activity itself. However, when EDTA is mixed with other antimicrobial agents, EDTA captures inorganic salts such as Ca ⁺² and Mg ⁺² , The structure is weakened and the penetration of the antimicrobial agent into the bacterial cell membrane is facilitated, so that the synergistic effect of the antibacterial activity can be demonstrated.

Claims (12)

A fatty acid tripeptide salt encapsulated in a lipid bilac,
Wherein the membrane of the lipid vesicle comprises lipid and ethanol,
Wherein the lipid comprises 5 to 20% (w / w) of cholesterol,
Wherein the lipid vesicle comprising the fatty acid tripeptide salt further comprises a chelating agent.
The method according to claim 1,
Wherein the fatty acid tripeptide salt is a palmitoyl tripeptide.
The method according to claim 1,
Wherein the lipid vesicle comprises a fatty acid tripeptide salt at a concentration of 1 to 4% (w / w).
delete delete The method according to claim 1,
Wherein the chelating agent comprises at least one member selected from the group consisting of EDTA, sodium polyphosphate, sodium metaphosphate, citric acid and gluconic acid.
The method according to claim 1,
Wherein the chelating agent is contained at 1/10 to 1/2 of the weight of the fatty acid tripeptide salt.
Preparing a solution in which a mixture of phospholipids and cholesterol is dissolved in ethanol (step a);
A step (b) of forming a hydrated liquid crystal phase by adding an aqueous solution in which the fatty acid tripeptide salt is dissolved to the solution of step (a) and dispersing the liquid crystal liquid phase to form lipid vesicle particles trapped with the fatty acid tripeptide salt,
Wherein the aqueous solution in which the fatty acid tripeptide salt is dissolved further comprises a chelating agent.
The method of claim 8,
Wherein the phospholipid and cholesterol are mixed at a weight ratio of 9.5 to 8.0: 0.5 to 2.
The method of claim 8,
Wherein the fatty acid tripeptide salt is a palmitoyl tripeptide.
delete The method of claim 8,
Wherein the chelating agent comprises at least one member selected from the group consisting of EDTA, sodium polyphosphate, sodium metaphosphate, citric acid and gluconic acid.

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