KR20240069119A - peptide for antimicrobial, anti-inflammatory or anti-acne and cosmetic composition for comprising the same - Google Patents

Abstract

The present invention relates to an antibacterial, anti-inflammatory or anti-acne peptide and a cosmetic composition containing the same. The peptide is an acyl group of a specific compound bound to the amine terminal of tryptophan, polytryptophan or valine-tryptophan dipeptide, and is an antibacterial, antibacterial, and anti-acne peptide. Since it has an inflammatory or anti-acne effect, it can be used as a cosmetic composition containing it.

Description

Antibacterial, anti-inflammatory or anti-acne peptide and cosmetic composition comprising the same {peptide for antimicrobial, anti-inflammatory or anti-acne and cosmetic composition for comprising the same}

The present invention relates to antibacterial, anti-inflammatory or anti-acne peptides and cosmetic compositions containing the same.

Acne is a Greek word meaning acne, which means acne vulgaris and comedo. It refers to a chronic sebaceous gland inflammatory disease in which a purulent reaction occurs in the pores and the structure of the skin is destroyed and damaged. The excretion action does not keep up with the production of sebum, and sebum remains in the pores, resulting in acne. It occurs only in the dormant part of the hair follicle. In most adolescent teenagers, when the secretion of androgen, a male hormone, increases most significantly, about 80% of people are infected and the sebaceous glands are infected. It commonly appears on the face, chest, and back, where it is widely distributed. Approximately 20% of adults show signs of acne, with more women than men, and Westerners who eat meat have a higher incidence of acne than Asians.

Among the various triggers that cause acne, a representative example is internal factors and excessive sebum secretion. Due to an imbalance in sex hormones, androgens promote sebum secretion and act as keratin thickening, and progesterone worsens acne after menstruation and adrenocortical hormones. In addition to secretion, secretion is also promoted by increased blood sugar levels. Acne is also promoted by lack of nutrients such as vitamins. Acne can also occur due to external stimuli, including stress, which is the most dangerous enemy of acne, as well as sunlight, which increases cell division and promotes cell division not only in the skin but also in the follicles, as well as season, climate, chemicals, diet, medicine, menstrual factors, and pregnancy. , birth control pills, cosmetics, pressure, friction, and picking with hands can be seen as factors that worsen acne symptoms.

From a medical perspective, the treatment of acne is primarily directed at suppressing the occurrence and progression of inflammation and the formation of microorganisms by administering antibiotics or corticosteroids, but this has side effects such as resistance to antibiotics, digestive problems, secondary infections, weight gain, and high blood pressure. There is a problem that may appear.

Meanwhile, from a cosmetic perspective, acne is managed through the use of cosmetics containing ingredients with anti-inflammatory, antibacterial, cleansing, exfoliating, and sebum controlling effects. However, the effect of existing cosmetics on improving acne is not significant, and there is a need for more research and development on cosmetics that can effectively improve acne-prone skin and prevent acne without irritating the skin.

Korean Patent No. 2115668.

The technical problem to be achieved by the present invention is to provide peptides for antibacterial, anti-inflammatory or anti-acne purposes.

In addition, the technical problem to be achieved by the present invention is to provide an antibacterial, anti-inflammatory or anti-acne cosmetic composition containing the above peptide.

The technical problem to be achieved by the present invention is not limited to the technical problem mentioned above, and other technical problems not mentioned can be clearly understood by those skilled in the art from the description below. There will be.

In order to achieve the above technical problem, an embodiment of the present invention includes a peptide selected from tryptophan, polytryptophan, and valine-tryptophan dipeptide, and the amine terminal of the peptide has an acyl functional group of the following formula (1): Combined, the polytryptophan provides an antibacterial, anti-inflammatory or anti-acne peptide in which 2 to 6 tryptophans are linked.

[Formula 1]

Here, R ¹ is C1~C14 alkyl, C2~C14 alkenyl, and C2~C14 alkynyl.

In an embodiment of the present invention, an (isopropyl methylphenoxy) acetyl group may be bonded to the amine terminal of the tryptophan.

In an embodiment of the present invention, an (isopropyl methylphenoxy) acetyl group may be bonded to the amine terminal of the polytryptophan.

In an embodiment of the present invention, the dipeptide may be one in which an (isopropyl methylphenoxy) acetyl group is bonded to the amine terminal of valine.

In an embodiment of the present invention, the bacteria may be Staphylococcus epidermidis or propionibacterium acnes.

In an embodiment of the present invention, the anti-inflammation may be due to inhibition of NO production or inhibition of iNOS expression.

In order to achieve the above technical problem, another embodiment of the present invention provides an antibacterial, anti-inflammatory or anti-acne cosmetic composition containing the Tide.

The present invention relates to an antibacterial, anti-inflammatory or anti-acne peptide and a cosmetic composition containing the same. The peptide is an acyl group of a specific compound bound to the amine terminal of tryptophan, polytryptophan or valine-tryptophan dipeptide, and is an antibacterial, antibacterial, and anti-acne peptide. Since it has an inflammatory or anti-acne effect, it can be used as a cosmetic composition containing it.

The effects of the present invention are not limited to the effects described above, and should be understood to include all effects that can be inferred from the configuration of the invention described in the description or claims of the present invention.

Figures 1 to 6 evaluate the antibacterial activity of IPMP-Ac-VW, a new peptide of the present invention, against acne bacteria.
Figures 7 and 8 evaluate the antibacterial activity of IPMP-Ac-VW, a new peptide of the present invention, against S. epidermidis.
Figures 9 and 10 evaluate the antibacterial activity of 2IP5MP-Ac-peptide and IPMP-AA, which are new peptides of the present invention, against acne bacteria.
Figures 11 and 12 confirm the cytotoxicity of the new peptides of the present invention, 2IP5MP-Ac-VW and 2IP5MP-Ac-WW, against HaCat cells (skin cells) and Raw 264.7 cells (immune cells), respectively.
Figures 13 to 16 confirm the anti-inflammatory effects of 2IP5MP-Ac-VW and 2IP5MP-Ac-WW, the new peptides of the present invention, respectively.

Hereinafter, the present invention will be described in detail.

The present invention relates to peptides for antibacterial, anti-inflammatory or anti-acne use.

The peptide of the present invention includes any one of tryptophan, polytryptophan, and valine-tryptophan dipeptide.

In the present invention, amino acids present in any peptide may be L-amino acids or D-amino acids. Preferably it may be L-amino acid.

The peptide has an acyl functional group (IPMP (Isopropyl methylphenoxy)-R ¹ CO-) of the following formula 1 attached to the amine terminal.

[Formula 1]

R ¹ of the acyl functional group may be C1~C14 alkyl, C2~C14 alkenyl, or C2~C14 alkynyl. R ¹ of the acyl functional group may vary depending on the type of compound used to introduce the acyl functional group by reacting with isopropyl methylphenol.

For example, if isopropyl methylphenol reacts with ethyl bromoacetate, the acyl functional group may be a 2-(isopropyl methylphenoxy)acetyl group.

The peptide is not limited thereto, but may be, for example, one in which an (isopropyl methylphenoxy) acetyl group is bonded to the amine terminal of the tryptophan. Additionally, an (isopropyl methylphenoxy) acetyl group may be bonded to the amine terminal of the polytryptophan. Additionally, the dipeptide may be one in which an (isopropyl methylphenoxy) acetyl group is bonded to the amine terminal of valine.

The polytryptophan is one in which 2 to 6 tryptophans are connected. Preferably, it may be 2 to 4 connected, and more preferably, it may be 2 to 3 connected.

The peptide according to one embodiment of the present invention can be synthesized using known means and under conditions known in the art. Methods for synthesizing the peptide include largely chemical synthesis and biological synthesis. For example, the peptide may be produced according to a chemical synthesis method, for example, a solid phase synthesis method. Biological synthesis methods include, for example, introducing a base sequence encoding a desired peptide into a protein expression vector, inducing expression in bacteria, and then isolating the protein using genetic engineering techniques.

In the present invention, 'antibacterial' may mean the effect of inhibiting the growth of microorganisms, such as bacteria related to skin diseases.

The bacteria may be, for example, Staphylococcus epidermidis or propionibacterium acnes, but are not limited thereto, and have the potential to cause skin contamination (disease). May contain microorganisms.

In the present invention, ‘anti-inflammatory’ may be used interchangeably with “inhibition or improvement of inflammation,” and anti-inflammation by the peptide of the present invention may be due, for example, to inhibition of NO production or inhibition of iNOS expression.

The present invention relates to an antibacterial, anti-inflammatory or anti-acne cosmetic composition containing the above peptide.

Specific descriptions related to the peptide and its efficacy are as described above.

For example, the composition may contain 0.1 μM to 1000 μM of the peptide, preferably 50 μM to 300 μM. If the composition contains too little of the peptide, it may not have sufficient skin antibacterial, anti-inflammatory, and anti-acne effects due to the above-mentioned peptide, and if it contains too much, it may have cytotoxicity. It is desirable to adjust appropriately.

The composition can be formulated into skin, lotion, toner, beauty soap, body wash, serum, cleansing lotion, essence, nutritional cream, pack, massage cream, etc., and other cosmetic products such as softening lotion, astringent lotion, nourishing lotion, and eye lotion. It can be formulated as cream, eye essence, cleansing foam, cleansing water, powder, body lotion, body cream, body oil, body essence, makeup base, foundation, shampoo or rinse, but is not limited to this.

Additionally, when used as a cosmetic composition, additional substances can be added to suit the formulation of the skin external agent or cosmetic. For example, but not limited to this, if the formulation is a paste, cream or gel, the carrier ingredients may include animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or Zinc oxide, etc. can be used, and when the formulation is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder can be used as a carrier ingredient. Especially, in the case of a spray, additional It may contain propellants such as chlorofluorohydrocarbons, propane/butane or dimethyl ether. Additionally, when the formulation is a solution or emulsion, a solvent, solubilizing agent, or emulsifying agent is used as a carrier component, preferably water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, or sorbitan fatty acid ester, but is not limited thereto. If the formulation is a suspension, the carrier ingredients include water, liquid diluents such as ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, Aluminum metahydroxide, bentonite, agar or tracant can be used, and when the formulation is a cleansing agent containing a surfactant, the carrier ingredients include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, Isethionate, imidazolinium derivatives, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative or ethoxylated Glycerol fatty acid ester, etc. may be used, but are not limited thereto.

Hereinafter, the present invention will be described in detail with reference to examples.

Manufacturing example

1. Synthesis of 2-(2-isopropyl-5-methylphenoxy)acetic acid

15.0 g (0.1 mol) of 2-Isopropyl-5-methylphenol and 27.6 g (0.2 mol) of potassium carbonate were added to 330 mL of acetone, followed by 18.4 g (0.11 mol) of ethyl bromoacetate and stirred at room temperature for 12 hours. After the reaction was filtered, 50 mL of distilled water was added to the obtained filtrate, and then 16.0 g (0.2 mol) of 50% sodium hydroxide solution was added and stirred at room temperature for 2 hours. After the reaction was completed, it was neutralized by adding 3M HCl and concentrated under reduced pressure. The resulting concentrate was dissolved in EA, washed with 0.1N HCl aqueous solution, and washed once with water. The organic layer was dried with magnesium sulfate and concentrated under reduced pressure to obtain 17.7 g (85%) of a solid compound.

2. Synthesis of H-(Trp(Boc))n-CTC resin

(a) H-Trp(Boc)-CTC resin

40 g (1.25 mmo/g) of 2-Chloro trityl chloride resin was placed in a reaction vessel, 300 mL of DCM was added, and the mixture was stirred for 30 minutes. The solution was removed, 300 mL of DMF was added, stirred for 30 minutes, and the solvent was removed again. Add 26.3 g (0.05 mol) of Fmoc-Trp(Boc)-OH, 300 mL of DMF, and 17.4 mL (0.1 mol) of DIPEA to the reaction vessel, react for 4 hours, filter, wash with 300 mL of DCM, and add MC:MeOH:DIPEA (17). :2:1) solution was added and reacted (capping) twice for 15 minutes each. Afterwards, the reaction solution was filtered and deprotection reaction was performed twice for 15 minutes each using 300 mL of 20% Piperidine/DMF solution. The reaction solution was filtered and washed three times with 300 mL of DMF and once with 300 mL of DCM.

(b) H- Trp(Boc)Trp(Boc)-CTC resin

To the H-Trp(Boc)-CTC resin synthesized in (a) above, 39.5 g (0.075 mol) of Fmoc-Trp(Boc)-OH, 11.1 g (0.825 mol) of HOBt, 300 mL of DMF, and 11.7 mL (0.075 mol) of DIC was added and reacted. After 4 hours of reaction, completion of the reaction was confirmed by Kaiser test, the reaction solution was removed by filtration, and then deprotection reaction was performed twice for 15 minutes each using 300 mL of 20% Piperidine/DMF solution. The reaction solution was filtered and washed three times with 300 mL of DMF and once with 300 mL of DCM to synthesize H-Trp(Boc)-Trp(Boc)-CTC resin.

(c) H- Trp(Boc)Trp(Boc)Trp(Boc)Trp(Boc)-CTC resin

To the H-Trp(Boc)Trp(Boc)-CTC resin synthesized in (b) above, 39.5 g (0.075 mol) of Fmoc-Trp(Boc)-OH, 11.1 g (0.825 mol) of HOBt, 300 mL of DMF, 11.7 mL of DIC (0.075 mol) was added and reacted. After 4 hours of reaction, completion of the reaction was confirmed by Kaiser test, the reaction solution was removed by filtration, and then deprotection reaction was performed twice for 15 minutes each using 300 mL of 20% Piperidine/DMF solution. The reaction solution was filtered and washed three times with 300 mL of DMF and once with 300 mL of DCM to synthesize H-Trp(Boc)-Trp(Boc)-CTC resin. After repeating the reaction of Fmoc-Trp(Boc)-OH with the synthesized H-Trp(Boc)Trp(Boc)Trp(Boc)-CTC resin once, dehydration was performed twice for 15 minutes each using 300 mL of 20% Piperidine/DMF solution. Through the protection reaction, H- Trp(Boc)Trp(Boc)Trp(Boc)Trp(Boc)-CTC resin was synthesized.

3. Synthesis of 2IP5MP-Ac-(Trp(Boc))n-CTC resin

To the H-(Trp(Boc))n-CTC resin synthesized in Preparation Example 2, 13.5 g (0.065 mol) of 2-(2-isopropyl-5-methylphenoxy)acetic acid synthesized in Preparation Example 1 and 11.4 g of HOBt (0.083 mol), 300 mL of DMF, and 11.7 mL (0.075 mol) of DIC were added and reacted under 300 mL of DMF. After 4 hours of reaction, completion of the reaction was confirmed by Kaiser test, and the reaction solution was filtered and washed three times with 300 mL of DMF and three times with 300 mL of DCM.

4. Synthesis of 2IP5MP-Ac-(Trp)n-OH

350 mL of 50% TFA/DCM was added to the 2IP5MP-Ac-(Trp(Boc))n-CTC resin synthesized in Preparation Example 3, and release and deprotection reactions were performed from the resin for 2 hours. The filtrate reacted twice was collected, concentrated, and dispersed in ether to recover crude 2IP5MP-Ac-(Trp)n-OH. The crude peptide was dissolved in distilled water, purified using prep-LC (column ID 8cm), and then freeze-dried to obtain a white powder.

2IP5MP-Ac-Trp-OH has the following formula (2), and the molecular weight of 11.5 g (purity: 97.5% (HPLC), yield: 58.5%) was determined by LC-MS to be 395.20 (M+1), (theoretical value: M = 394.19).

[Formula 2]

2IP5MP-Ac-TrpTrp-OH has the following formula (3), and the molecular weight of 14.3g (purity: 96.5% (HPLC), yield 49.3%) was determined by LC-MS to be 581.24 (M+1), (theoretical value: M = 580.27).

[Formula 3]

2IP5MP-Ac-TrpTrpTrpTrt-OH has the following formula 4, and the molecular weight of 24.5 g (purity: 97.4% (HPLC), yield: 51.4%) was determined by LC-MS to be 953.45 (M+1), (theoretical value: M = 952.43).

[Formula 4]

5. Synthesis of 2IP5MP-Ac-(Val)-(Trp)-OH

(a) H-Val-Trp(Boc)-CTC resin

25.5 g (0.075 mol) of Fmoc-Val-OH, 11.1 g (0.825 mol) of HOBt, 300 mL of DMF, and 11.7 mL (0.075 mol) of DIC to the H-Trp(Boc)-CTC resin synthesized in Preparation Example 2(a). was added and reacted. After 4 hours of reaction, completion of the reaction was confirmed by Kaiser test, the reaction solution was removed by filtration, and then deprotection reaction was performed twice for 15 minutes each using 300 mL of 20% Piperidine/DMF solution. The reaction solution was filtered and washed three times with 300 mL of DMF and once with 300 mL of DCM to synthesize H-Val-Trp(Boc)-CTC resin.

(b) 2IP5MP-Ac-Val-Trp(Boc)-CTC resin

11.4 g (0.083 mol) of HOBt and 300 mL of DMF were added to the H-Val-Trp(Boc)-CTC resin synthesized in (a) above, and then 2-(2-isopropyl-5-methylphenoxy)acetic synthesized in Preparation Example 1. 13.5g (0.065mol) of acid and 11.7mL (0.075mol) of DIC were added and reacted. After 4 hours of reaction, completion of the reaction was confirmed by Kasiser test, and the reaction solution was filtered out and washed three times with 300 mL of DMF and three times with 300 mL of DCM.

(c) Synthesis of 2IP5MP-Ac-Val-Trp-OH

350 mL of 50% TFA/DCM was added to the 2IP5MP-Ac-Val-Trp(Boc)-CTC resin synthesized in (b) above, and the release and deprotection reactions were performed from the resin for 30 minutes. The filtrate from two reactions was collected, concentrated, and dispersed in ether to recover crude 2IP5MP-Ac-Val-Trp-OH. The crude peptide was dissolved in distilled water, purified using prep-LC (column ID 8cm), and then freeze-dried to obtain 15.1 g of white powder (purity: % (HPLC), yield: 61.2%), which was obtained by LC-MS. The molecular weight was measured as (M+1), (theoretical value: M = 493.26).

[Formula 5]

Experiment example

1. Evaluation of antibacterial efficacy of new peptides

Antibacterial efficacy against P. acnes and S. epidermidis was evaluated using the test substances shown in Table 1 below. Specifically, P. acnes was cultured in Reinforced Clostridial Medium (RCM) at 37°C, 1.5x10 ⁶ CFU/mL, 96 well plate for 72 hours (simultaneous material treatment), anaerobic condition, and S. epidermidis was cultured in Nutrient Medium. Culture was carried out at 37°C, 1.5x10 ⁶ CFU/mL, 96 well plate for 24 hours (simultaneous material treatment), aerobic conditions. The test method was described by Ryu, AR., Han, CS., and Oh, HK. et al. Chlorin e6-mediated photodynamic inactivation with halogen light against microbes and fungus. Toxicol. Environ. Health Sci. 7, 231-238 (2015). Refer to https://doi.org/10.1007/s13530-015-0243-z.

test substance note IPMP-Ac-VW 2IP5MP-Ac-VW
(MW 493.60) 4IP3MP-Ac-VW
(MW 493.60) 5IP2MP-Ac-VW (MW 493.60) IPMP-Ac-KVK 2IP5MP-Ac-KVK
(MW 563.74) 4IP3MP-Ac-KVK
(MW 563.74) 5IP2MP-Ac-KVK (MW 563.74) IPMP-Ac-GHK 2IP5MP-Ac-GHK
(MW 530.63) 4IP3MP-Ac-GHK
(MW 530.63) 5IP2MP-Ac-GHK (MW 530.63) 2IP5MP (MW 150.22) comparison group 4IP3MP (MW 150.22) comparison group 5IP2MP (MW 150.22) comparison group VW (MW 303.36) comparison group KVK (MW 373.50) comparison group GHK (MW 340.38) comparison group Erythromycin (MW 733.93) Positive control group

Here, V, W, G, H, and K are abbreviations for amino acids, which are V (valine), W (tryptophan), G (glycine), H (histidine), and K (lysine). IPMP is an (Isopropyl methylphenoxy)acetyl group. For example, 2IP5MP-Ac-VW is 2-(2-isopropyl-5-methylphenoxy)acetyl-VW, and 4IP3MP-Ac-VW is 2-(4-isopropyl-3- It is methylphenoxy)acetyl-VW.

As a result, IPMP-Ac-VW showed excellent antibacterial activity against acne bacteria (Figures 1, 5, and 6). It showed antibacterial activity of around 50% at a concentration of 125 μM and 70-80% at a concentration of 250 μM. In addition, the concentration (IC ₅₀ ) that inhibits acne bacteria growth by 50% was 130 μM for 2IP5MP-Ac-VW, 164 μM for 4IP3MP-Ac-VW, and 146 μM for 5IP2MP-Ac-VW (Figure 2). In comparison, IPMP-Ac-KVK and IPMP-Ac-GHK had very little antibacterial activity against acne bacteria (Figures 3 and 4).

Additionally, under these experimental conditions, VW, KVK, and GHK themselves showed little antibacterial activity against acne bacteria. In this experimental condition, 2IP5MP, 4IP3MP, and 5IP2MP themselves had minimal antibacterial activity against acne bacteria. 125μM of 2IP5MP, 4IP3MP, and 5IP2MP showed an antibacterial activity against acne bacteria of about 10%, and 250μM of 2IP5MP, 4IP3MP, and 5IP2MP showed an antibacterial effect against acne bacteria of about 20%.

IPMP-Ac-VW also showed antibacterial activity against S. epidermidis, with an antibacterial activity of approximately 30% at a concentration of 250 μM (Figures 7 and 8).

2. Evaluation of antibacterial activity against acne bacteria of 2IP5MP-Ac-W, 2IP5MP-Ac-WW, and 2IP5MP-Ac-WWWW

Based on 2IP5MP-Ac-VW, the antibacterial activity against acne bacteria of 2IP5MP-Ac-W, 2IP5MP-Ac-WW, and 2IP5MP-Ac-WWWW was compared at three concentrations (62.5 μM, 125 μM, 250 μM). The antibacterial efficacy evaluation test method was the same as Experiment 1 above.

The test substances are listed in Table 2 below.

test substance note 2IP5MP-Ac-Peptide 2IP5MP-Ac-VW 2IP5MP-Ac-W 2IP5MP-Ac-WW 2IP5MP-Ac-WWWW Acetic Acid Intermediates 4IP3MP-AA 5IP2MP-AA 2IP5MP-AA GA-VW

As a result, among the 2IP5MP-based peptides, the antibacterial activity of 2IP5MP-Ac-VW and 2IP5MP-Ac-W against P. acnes was similar at three concentrations. The antibacterial activity of 2IP5MP-Ac-WW against P. acnes showed better results than 2IP5MP-Ac-VW (or 2IP5MP-Ac-W) at three concentrations. 2IP5MP-Ac-WWWW did not show a concentration-dependent change in antibacterial activity, but it was found to have antibacterial activity (Figure 9).

In contrast, the three AA intermediates and GA-VW did not show significant changes in antibacterial activity at the three concentrations (Figure 10).

3. Evaluation of cytotoxic and anti-inflammatory efficacy

(1) Cytotoxicity evaluation

MTT assay was performed using 2IP5MP-Ac-VW and 2IP5MP-Ac-WW serially diluted 2-fold at 500 μM. RAW 264.7 (macrophage-derived) cells and HaCaT cells (skin keratinocytes) were treated with test substances to confirm cell survival.

As a result, 2IP5MP-Ac-WW had stronger cytotoxicity than 2IP5MP-Ac-VW in two types of cells, skin cells and immune cells. 2IP5MP-Ac-WW shows greater potential for skin irritation than 2IP5MP-Ac-VW. This is believed to be directly related to the result that 2IP5MP-Ac-WW has superior antibacterial activity against acne bacteria than 2IP5MP-Ac-VW (Figures 11 and 12)

(2) Evaluation of anti-inflammatory efficacy

RAW 264.7 (macrophage-derived) cells were treated with test substances after inducing inflammation with LPS.

NO measurement was performed using the Griess reagent using test substances serially diluted 2-fold at 500 μM of 2IP5MP-Ac-VW and 2IP5MP-Ac-WW.

Inhibition of inflammatory factor expression was measured by Western transfer for 2IP5MP-Ac-VW and 2IP5MP-Ac-WW at non-cytotoxic concentrations, using iNOS, COX-2, and IL-1β antibodies as primary antibodies. , the ability to suppress IL-1β expression was evaluated.

As a result, 2IP5MP-Ac-VW and 2IP5MP-Ac-WW inhibited LPS-induced NO production and iNOS expression at similar levels in raw cells, but did not inhibit COX-2 and IL-1β. These two peptides are believed to exhibit anti-inflammatory effects through inhibition of NO production and iNOS expression among various inflammatory mechanisms (Figures 13 to 16).

4. Formulation evaluation

(1) Peptide solution manufacturing

The new peptides of the present invention show poor solubility in pure water, so it is difficult to add them directly into the solubilized formulation. For convenience of weighing and manufacturing, the process was carried out by first dissolving them in polyol at a high concentration and then adding them into the formulation. Table 3 below shows the solution of three types of peptides among the peptides prepared above at 1,000 ppm each.

Peptide A
Solution Peptide B
Solution Peptide C
Solution Water 78.350 78.350 78.350 1,2-Hexanediol 1.500 1.500 1.500 Ethylhexylglycerin 0.050 0.050 0.050 Dipropylene Glycol 20.000 20.000 20.000 IPMP-Ac-VW (2IP5MP-Ac-VW) 0.100 2IP5MP 0.100 VW 0.100 100.000 100.000 100.000

(2) Ampoule formulation

1) Manufacturing method

An ampoule formulation was prepared by a commonly known method, and the new peptide of the present invention was introduced into the final manufacturing process as a 1,000 ppm solution.

As shown in Table 4, Examples 1 and 2 contain 35 ppm (= 70 uM) and 65 ppm (= 131 uM) of 2IP5MP-Ac-VW in the final formulation, respectively, and Comparative Example 1 is a formulation without peptide added. Example 2 contains 65 ppm (= 131 um) of 2IP5MP, and Comparative Example 3 contains 65 ppm (= 131 uM) of VW.

INCI Name Example-1 Example-2 Comparative Example-1 Comparative example-2 Comparative example-3 Water 78.200 75.200 81.700 75.200 75.200 Butylene Glycol 7.000 7.000 7.000 7.000 7.000 Glycerin 4.000 4.000 4.000 4.000 4.000 Propanediol 4.000 4.000 4.000 4.000 4.000 1,2-Hexanediol 1.500 1.500 1.500 1.500 1.500 Panthenol 0.500 0.500 0.500 0.500 0.500 Hydrolyzed Collagen 0.300 0.300 0.300 0.300 0.300 Polyglyvcerin-4 Caprate 0.300 0.300 0.300 0.300 0.300 Carbomer 0.150 0.150 0.150 0.150 0.150 Tromethamine 0.150 0.150 0.150 0.150 0.150 Allantoin 0.100 0.100 0.100 0.100 0.100 Sodium Polyacryloydimethyl Taurate 0.100 0.100 0.100 0.100 0.100 Ethylhexylglycerin 0.050 0.050 0.050 0.050 0.050 Adenosine 0.040 0.040 0.040 0.040 0.040 Disodium EDTA 0.030 0.030 0.030 0.030 0.030 Sodium Hyaluronate 0.030 0.030 0.030 0.030 0.030 Fragrance 0.050 0.050 0.050 0.050 0.050 Peptide A Solution (1,000 ppm) 3.500 6.500 Peptide B Solution (1,000 ppm) 6.500 Peptide C Solution (1,000 ppm) 6.500 100.000 100.000 100.000 100.000 100.000

2) Formulation stability evaluation

The room temperature and high temperature stability of the ampoule formulation containing the above peptide was confirmed.

Cosmetic formulations manufactured according to cosmetic standards such as appearance, pH, and viscosity were evaluated over time, and it was confirmed that there was no significant difference overall (Table 5). This confirms that the contained peptides are stable without any interaction with the main components (thickener, polyol, etc.) that make up the ampoule formulation, so it is expected that there will be no major difficulties in using it in the ampoule formulation.

INCI Name Example-1 Example-2 Comparative example-1 Comparative example-2 Comparative example-3 25 degrees 4 weeks O/O/O O/O/O O/O/O O/O/O O/O/O 25 degrees 8 weeks O/O/O O/O/O O/O/O O/O/O O/O/O 25 degrees 12 weeks O/O/O O/O/O O/O/O O/O/O O/O/O 45 degrees 4 weeks O/O/O O/O/O O/O/O O/O/O O/O/O 45 degrees 8 weeks O/O/O O/O/O O/O/O O/O/O O/O/O 45 degrees 12 weeks O/O/O O/O/O O/O/O O/O/O O/O/O

* Evaluation: Appearance / pH / Viscosity

(3) Cream formulation

1) Manufacturing method

A cream formulation was prepared by a commonly known method, and the new peptide of the present invention was introduced into the final manufacturing process as a 1,000 ppm solution.

As shown in Table 6, Examples 3 and 4 contain 35 ppm (= 70 uM) and 65 ppm (= 131 uM) of 2IP5MP-Ac-VW, respectively, in the final formulation, and Comparative Example 4 is a formulation without peptide added, Comparative Example. 5 contains 65 ppm (= 131 um) of 2IP5MP, and Comparative Example 6 contains 65 ppm (= 131 uM) of VW.

INCI Name Example-3 Example-4 Comparative example-4 Comparative example-5 Comparative example-6 Water 70.180 67.180 73.680 67.180 67.180 Glycerin 5.000 5.000 5.000 5.000 5.000 Butylene Glycol Dicaprylate/Dicaprate 5.000 5.000 5.000 5.000 5.000 Triethylhexanoin 3.000 3.000 3.000 3.000 3.000 Butylene Glycol 3.000 3.000 3.000 3.000 3.000 Squalane 2.000 2.000 2.000 2.000 2.000 Cetearyl Olivate 2.100 2.100 2.100 2.100 2.100 1,2-Hexanediol 1.500 1.500 1.500 1.500 1.500 Sorbitan Olivate 1.400 1.400 1.400 1.400 1.400 Behenyl Alcohol 0.600 0.600 0.600 0.600 0.600 Panthenol 0.500 0.500 0.500 0.500 0.500 Behenyl Alcohol 0.500 0.500 0.500 0.500 0.500 Arachidyl Alcohol 0.300 0.300 0.300 0.300 0.300 Carbomer 0.300 0.300 0.300 0.300 0.300 Tromethamine 0.300 0.300 0.300 0.300 0.300 Hydrolyzed Collagen 0.200 0.200 0.200 0.200 0.200 Arachidyl Glucoside 0.100 0.100 0.100 0.100 0.100 Allantoin 0.100 0.100 0.100 0.100 0.100 Sodium Polyacryloyldimethyl Taurate 0.050 0.050 0.050 0.050 0.050 Ethylhexylglycerin 0.050 0.050 0.050 0.050 0.050 Sodium Hyaluronate 0.050 0.050 0.050 0.050 0.050 Adenosine 0.040 0.040 0.040 0.040 0.040 Disodium EDTA 0.030 0.030 0.030 0.030 0.030 Fragrance 0.200 0.200 0.200 0.200 0.200 Peptide A Solution (1,000 ppm) 3.500 6.500 Peptide B Solution (1,000 ppm) 6.500 Peptide C Solution (1,000 ppm) 6.500 100.000 100.000 100.000 100.000 100.000

2) Formulation stability evaluation

The room temperature and high temperature stability of the cream formulation containing the above peptide was confirmed.

Cosmetic formulations manufactured according to cosmetic standards such as appearance, pH, and viscosity were evaluated over time, and it was confirmed that there was no significant difference overall (Table 7). This confirms that the contained peptides are stable without any interaction with the main ingredients (thickener, polyol, etc.) that make up the cream formulation, so it is expected that there will be no major difficulties in using it in cream formulations.

INCI Name Example-3 Example-4 Comparative example-4 Comparative example-5 Comparative example-6 25 degrees 4 weeks O/O/O O/O/O O/O/O O/O/O O/O/O 25 degrees 8 weeks O/O/O O/O/O O/O/O O/O/O O/O/O 25 degrees 12 weeks O/O/O O/O/O O/O/O O/O/O O/O/O 45 degrees 4 weeks O/O/O O/O/O O/O/O O/O/O O/O/O 45 degrees 8 weeks O/O/O O/O/O O/O/O O/O/O O/O/O 45 degrees 12 weeks O/O/O O/O/O O/O/O O/O/O O/O/O

* Evaluation: Appearance / pH / Viscosity

Claims (7)

Contains any one of tryptophan, polytryptophan, and valine-tryptophan dipeptide,
An acyl functional group of the following formula (1) is bound to the amine terminal of the peptide,
The polytryptophan is an antibacterial, anti-inflammatory or anti-acne peptide in which 2 to 6 tryptophans are linked.
[Formula 1]

Here, R ¹ is C1~C14 alkyl, C2~C14 alkenyl, and C2~C14 alkynyl. The peptide according to claim 1, wherein an (isopropyl methylphenoxy) acetyl group is bonded to the amine terminal of the tryptophan. The peptide according to claim 1, wherein an (isopropyl methylphenoxy) acetyl group is bonded to the amine terminal of the polytryptophan. The method according to claim 1, wherein the dipeptide is an antibacterial, anti-inflammatory or anti-acne peptide in which an (isopropyl methylphenoxy) acetyl group is bonded to the amine terminal of valine. The peptide according to claim 1, wherein the bacteria is Staphylococcus epidermidis or propionibacterium acnes. The antibacterial, anti-inflammatory or anti-acne peptide according to claim 1, wherein the anti-inflammatory is achieved by inhibiting NO production or iNOS expression. A cosmetic composition for antibacterial, anti-inflammatory or anti-acne use comprising the peptide of any one of claims 1 to 6.