KR102540535B1 - Antimicrobial peptide and uses thereof - Google Patents

Abstract

The present invention relates to an antibacterial peptide derivative, and a cosmetic composition for improving acne containing the peptide derivative as an active ingredient can exhibit excellent antibacterial effects.

Description

Antimicrobial peptide and uses thereof {Antimicrobial peptide and uses thereof}

The present invention relates to an antibacterial peptide derivative and its use, and specifically to a cosmetic composition for improving acne containing the peptide derivative as an active ingredient.

Acne is a chronic inflammatory disease of the hairy sebaceous gland unit, and various skin changes such as comedones (sebum accumulated in hair follicles and hardened), papules (raised skin lesions less than 1 cm in size), pus blisters, nodules, and pseudocysts appear. As a result of this, concave scars or enlarged scars may be left. It often occurs on the face, neck, and chest where sebaceous glands are concentrated.

The exact cause of acne is not known, but in general, acne is caused by excessive sebum secretion, excessive skin keratinization, and free fatty acid and various small molecules produced by the lipase of Cutibacterium acnes, an anaerobic skin flora. An inflammatory reaction caused by stimulation of substances is known to be the cause. To treat acne, oral or external medications have been used. For example, antiandrogens that inhibit sebum production, non-steroidal anti-inflammatory drugs that act as anti-inflammatory agents, resochinol, sulfur, salicylic acid, benzoyl peroxide, Antibacterial agents such as isotretinoin, keratin exfoliants, and antibiotics such as tetracycline, erythromycin, and mecrocycline inhibit the activity of Cutibacterium acnes.

However, although the acne treatment shows a certain degree of effect, the need for the development of products that are safe for the human body and have excellent effects on acne skin due to side effects, ease of use, and needs for more improved effects is steadily emerging.

Recently, a polymer of amino acids, which is structurally simpler than proteins, but has antibacterial effects, has been developed for cosmetic materials classified into various peptides, such as vegetable peptides, synthetic peptides, and microorganism-derived peptides.

These antimicrobial peptides are amino acid polymers composed of 2 to 50 amino acids, and are mainly positively charged basic amino acid polymers. These antimicrobial peptides are biological defense factors possessed by almost all living organisms, including bacteria, plants, vertebrates and invertebrates, and antimicrobial peptides are the only external microorganisms, especially for invertebrates such as insects and crustaceans that do not have an immune system. It is a defense system against

These antimicrobial peptides have a mechanism of action that physically destroys cell membranes of microorganisms, and thus can provide sufficient defense against strains resistant to antibiotics due to misuse and abuse of antibiotics.

In addition, antibacterial peptides are fast, effective, and have a wide antibacterial spectrum, so that they can exhibit excellent antibacterial effects, and can be easily mass-produced and thus manufactured at a relatively low production cost.

Therefore, it is necessary to study low molecular weight antibacterial peptides that are safe and harmless to the human body, have improved antibacterial effects, and are easy to mass-produce.

Republic of Korea Patent Publication 10-2022-0090857 A Republic of Korea Patent Publication 10-2020-0006492 A

An object of the present invention is to provide an antimicrobial peptide derivative having excellent antibacterial effect and safety.

Another object of the present invention is to provide a cosmetic composition for improving acne containing the antibacterial peptide derivative as an active ingredient.

The present invention provides an antimicrobial peptide derivative represented by Formula 1 below.

[Formula 1]

A ₁ -KRKR

[In Formula 1,

K is Lys;

R is Arg;

A ₁ is a C12-C24 fatty acid.]

A ₁ of Formula 1 may be specifically palmitoyl.

In addition, the present invention provides an antimicrobial peptide derivative represented by Formula 2 below.

[Formula 2]

A ₂ -KWKLFKK

[In Formula 2,

K is Lys;

W is Trp;

L is Leu;

F is Phe;

A ₂ is a C12-C24 fatty acid.]

A ₂ of Formula 2 may be specifically palmitoyl.

The present invention provides a cosmetic composition for improving acne containing an antibacterial peptide derivative according to an embodiment of the present invention as an active ingredient.

In addition, the cosmetic composition for improving acne may be in the form of lotion, toner, lotion, nutrient lotion, nutrient cream, massage cream, essence, pack, or microneedle patch.

The antibacterial peptide derivative of the present invention can exhibit excellent antibacterial effect and high human safety. In addition, since it contains a smaller number of amino acid sequences than natural antibacterial peptides, it can be easily prepared by a simple process, so it has high economic efficiency and can be easily mass-produced.

In addition, the cosmetic composition for improving acne containing the antibacterial peptide derivative of the present invention as an active ingredient can significantly improve acne improvement due to its excellent antibacterial effect, and can solve the problems of cytotoxicity and skin allergy.

Hereinafter, the antibacterial peptide derivative of the present invention and the cosmetic composition for improving acne containing the same as an active ingredient will be described in detail.

The singular form used in the present invention may be intended to include the plural form as well, unless the context specifically dictates otherwise.

Further, as used herein, numerical ranges include lower and upper limits and all values within that range, increments logically derived from the shape and breadth of the defined range, all values defined therebetween, and the upper limit of the numerical range defined in a different form. and all possible combinations of lower bounds. Unless otherwise specifically defined in the specification of the present invention, values outside the numerical range that may occur due to experimental errors or rounding of values are also included in the defined numerical range.

As described in the present invention, "comprising" is an open-ended description having the same meaning as expressions such as "comprises", "includes", "has" or "characterized by", elements not additionally listed, No materials or processes are excluded.

Hereinafter, the present invention will be described in detail. At this time, if there is no other definition in the technical terms and scientific terms used, they have meanings commonly understood by those of ordinary skill in the art to which this invention belongs, and will unnecessarily obscure the gist of the present invention in the following description. Descriptions of possible known functions and configurations are omitted.

The term “antibacterial peptide” as used herein refers to a polymer composed of amino acids linked by amide bonds or peptide bonds, and has growth inhibitory activity against microorganisms.

Amino acid sequences described in the present invention are abbreviated according to the IUPAC-IUB nomenclature. Specifically, alanine is A (Ala), arginine is R (Arg), asparagine is N (Asn), aspartic acid is D (Asp), cysteine is C (Cys), glutamic acid is E (Glu), and glutamine is Q (Gln). ), Glycine is G (Gly), Histidine is H (His), Isoleucine is I (Ile), Leucine is L (Leu), Lysine is K (Lys), Methionine is M (Met), Phenylalanine is F (Phe ), proline was described as P (Pro), serine as S (Ser), threonine as T (Thr), tryptophan as W (Trp), tyrosine as Y (Tyr), and valine as V (Val). Amino acids of the antimicrobial peptide may have either L or D conformation.

The peptide derivative according to the present invention can be used without limitation as long as it is a conventional peptide synthesis method known in the art. chemical synthesis methods such as, for example, solution phase peptide synthesis, solid-phase peptide synthesis, fragment condensation and F-moc or T-boc chemistries; And a biological synthesis method prepared by expressing a polynucleotide encoding an antimicrobial peptide in genetically engineered E. coli; it may be synthesized by a solid phase peptide synthesis method, but is not limited thereto.

The solid-phase peptide synthesis method includes the steps of a) loading an amino acid protected with a protecting group into a resin; b) removing a protecting group from an amino acid; c) performing a coupling reaction with an amino acid; d) confirming the completion of the reaction; e) removing the resin and protecting groups; and f) solidifying the peptide.

In the solid phase peptide synthesis method, steps b), c) and d) may be repeated according to the number of desired amino acids, and the resin in step a) is Wang resin, chlorotrityl resin ), polystyrene resine, etc., and can be used by swelling the resin by adding an appropriate solvent to the resin before use. Amino acids may be bound to the resin by reacting the swollen resin with an amino acid protected by a protecting group using diisopropylcarbodiimide (DIC), N,N-diisopropylethylamine (DIPEA), 4-dimethylaminopyridine (DMAP), or a mixture thereof. .

Step b) is a step of removing a protecting group from an amino acid, and is not particularly limited as long as it is a method generally known in the art. can be removed. The protecting group of the amino acid may include Fmoc (9-Fluorenylmethoxycarbonyl), but is not limited thereto.

The coupling agent used in step c) is not limited as long as it is generally used in the art, and for example, HOBt-DCC (N-Hydroxybenzotriazole-Dicyclohexylcarbodiimide) or HOBt-DIC (N-Hydroxybenzotriazole-Diisopropylcarbodiimide) can be used. In addition, Dicyclohexylcarbodiimide (DCC), Diisopropylcarbodiimide (DIC), Benzotriazol-1-yl-oxy-tripyrrolidinophosphonium hexafluorophosphate (PyBOP), Benzotriazole-1-yl-oxy-tris-(dimethylamino)-phosphonium hexafluorophosphate (BOP), 1-[ From Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), N-Hydroxybenzotriazole (HOBt) and 1-Hydroxy-7-azabenzotriazole (HOAt) It may be one or two or more selected, and an organic acid or organic base such as tirfluoroacetic acid (TFA), N,N-Diisopropylethylamine, and N-methymorpholine may be added.

Step d) is a step of performing a Kaiser test to confirm completion of the reaction, and is a process of confirming whether blue color appears depending on the presence of primary amine using ninhydrin as a small amount of resin. When it appears blue, it is determined that the unreacted portion remains, and the re-coupling reaction can proceed.

Step e) is a step of removing the resin and the protecting group, and is not limited as long as it is a method generally used in the art, specifically selected from Tirfluoroacetic acid (TFA), Triisopropylsilane (TIS), Thioanisole, Ethanedithiol (EDT) and water. It can be carried out by adding a cocktail consisting of one or two or more.

Step f) is a step of solidifying the peptide, and a precipitated solid may be produced by adding an excessive amount of diethyl ether solvent, but is not limited thereto.

In the method for producing a peptide derivative according to the present invention, after step d) is completed, a reaction of introducing each of the peptides using a fatty acid may be additionally performed. The antimicrobial peptide derivative of the present invention can be obtained by proceeding with steps e) and f) with the reactant into which the prepared fatty acid is introduced.

The antimicrobial peptide derivative according to the present invention is a peptide having a sequence of 4 or 7 amino acids coupled to a fatty acid, and can exhibit an antibacterial effect equal to or higher than that of a natural antimicrobial peptide, and is therefore easy to manufacture and suitable for mass production.

The present invention provides an antimicrobial peptide derivative represented by Formula 1 below.

[Formula 1]

A ₁ -KRKR

[In Formula 1,

K is Lys;

R is Arg;

A ₁ is a C12-C24 fatty acid.]

A ₁ of Formula 1 may be a C14-C18 fatty acid, and specifically may be palmitoyl.

In detail, the antimicrobial peptide derivative represented by Formula 1 may be represented by Formula 3 below.

[Formula 3]

[In Formula 3,

A ₁ is a C12-C24 fatty acid.]

A ₁ of Formula 3 may be a C14-C18 fatty acid, and specifically may be palmitoyl.

In addition, the present invention provides an antimicrobial peptide derivative represented by Formula 2 below.

[Formula 2]

A ₂ -KWKLFKK

[In Formula 2,

K is Lys;

W is Trp;

L is Leu;

F is Phe;

A ₂ is a C12-C24 fatty acid.]

A ₂ of Formula 2 may be a C14-C18 fatty acid, and specifically may be palmitoyl.

In detail, the antimicrobial peptide derivative represented by Formula 2 may be represented by Formula 4 below.

[Formula 4]

[In Formula 4,

A ₂ is a C12-C24 fatty acid.]

A ₂ of Formula 4 may be a C14-C18 fatty acid, and specifically may be palmitoyl.

As A ₁ and A ₂ of Formulas 1 to 4 contain an acyl group derived from a C14-C18 fatty acid, more specifically a palmitoyl group, they may have characteristics of a lipopeptide, and the antibacterial activity of the antimicrobial peptide is significantly improved. can This is opposite to the tendency of antibacterial activity to rather decrease as general antibacterial peptides contain a long-chain acyl group, and is interpreted as a result of the combination of a KRKR sequence or a KWKLFKK sequence and a long-chain acyl group. Accordingly, antibacterial peptides containing an acyl group derived from C14-C18 fatty acids exhibit remarkable antibacterial activity and low cytotoxicity, which is a significant advantage for minimal inhibitory concentration (MIC) in applications requiring antibacterial and safety, such as cosmetic compositions, pharmaceutical compositions, and food compositions can provide.

The present invention provides a cosmetic composition for improving acne containing an antibacterial peptide derivative according to an embodiment of the present invention as an active ingredient.

The cosmetic composition for improving acne may have antibacterial activity against gram-positive bacteria or gram-negative bacteria.

The gram-positive bacteria may be one or two or more selected from Cutibacterium acnes, Staphylococcus epidermidis, and Staphylococcus aureus, and the gram-negative bacteria may be Escherichia coli (Escherichia coli) and Pseudomonas aeruginosa (Pseudomonas aeruginosa) may be selected from one or two or more.

In addition, the Gram-positive bacteria are Staphylococcus aureus, Streptococcus agalactiae, Streptococcus pneumoniae, Corynebacterium diphtheriae, Bacillus subtilis (Bacillus subtilis) and Enterococcus faecium (Enterococcus faecium) may further include one or more selected from, the Gram-negative bacteria Escherichia coli (Escherichia coli), Salmonella typhimurium (Salmonella typhimurium), Shigella dicente It may further include one or two or more selected from Shigella dysenteriae and Vibrio cholerae.

The cosmetic composition for improving acne exhibits excellent antibacterial activity by including the antibacterial peptide derivative according to an embodiment of the present invention as an active ingredient, and in particular, can exhibit excellent antibacterial activity against the acne bacterium Cutibacterium acnes. It can show improved acne improvement effect. In addition, since the antimicrobial peptide derivative according to one embodiment has low cytotoxicity to human-derived cells, it can be used as a raw material for cosmetics for improving acne with improved safety and lower harm to the human body than conventional antibacterial peptides of natural origin.

The active ingredient included in the cosmetic composition for improving acne may be added at a concentration of 10 to 300 μg/mL.

The cosmetic composition for improving acne according to an embodiment may be a lotion, toner, lotion, nutrient lotion, nutrient cream, massage cream, essence, pack or microneedle patch formulation, and in detail, softening lotion, astringent lotion, nutrient lotion, Nourishing cream, massage cream, eye cream, eye essence, essence, cleansing cream, cleansing lotion, cleansing foam, cleansing water, mask pack, powder, body lotion, body cream, body essence, body cleanser, sunscreen cream, hair dye, shampoo , It may have one or two or more formulations selected from rinses, toothpaste, mouth fresheners, hairdressing agents, hair growth agents, lotions, ointments, gels, creams, patches, and sprays.

When the formulation of the present invention is a paste, cream or gel, carrier components selected from animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc and zinc oxide One or more may be used.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be further used as a carrier component, especially in the case of a spray, additionally chlorofluorohydrocarbon, propellants such as propane/butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent may be further used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene Glycol, 1,3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycol or fatty acid esters of sorbitan may be included therein.

When the formulation of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol as a carrier component; A suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester or polyoxyethylene sorbitan ester may be further used, and crystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanthate may also be added. can be used as

When the formulation of the present invention is surfactant-containing cleansing, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters may be used.

The cosmetic composition for improving acne according to an embodiment of the present invention may further include additional additives as desired, which include, for example, a stabilizer, an emulsifier, a thickener, a moisturizer, a liquid crystal film strengthening agent, a pH adjuster, one or more aqueous additives selected from pH adjusters, water-soluble polymers, coating agents, metal ion sequestering agents, organic amines, polymer emulsions, skin nutrients, antioxidants, antioxidant aids, preservatives, fragrances, and the like; and one or more oily additives selected from oils, waxes, hydrocarbon oils, higher fatty acid oils, higher alcohols, synthetic ester oils, silicone oils, and the like.

The cosmetic composition for improving acne of the present invention may include two or more different types of antibacterial peptide derivatives selected from peptide derivatives according to an embodiment, and a synergistic effect is evoked than when one type is included alone, resulting in a more improved antibacterial effect can represent

The microneedle patch includes a microneedle unit in which a plurality of microneedles containing an antimicrobial peptide derivative according to an embodiment are positioned; and a base portion coupled to the microneedle portion.

Hereinafter, the antibacterial peptide derivative according to the present invention and the cosmetic composition for improving acne containing the same as an active ingredient will be described in more detail through specific examples.

However, the following examples are only one reference for explaining the present invention in detail, but the present invention is not limited thereto, and may be implemented in various forms. Also, the terms used in the description in the present invention are only for effectively describing specific embodiments, and are not intended to limit the present invention.

[Example 1] Preparation of Compound 1

Example 1 (Compound 1) was synthesized according to solid-phase peptide synthesis. Specifically, amine resin was used, and for each peptide, Arg, Lys, Arg, Lys, which are amino acids using Fmoc (9-fluorenylmethoxycarbonyl) as an N-terminal protecting group, are used for the coupling reaction in order, followed by palmitic acid for the coupling reaction was used for For each of the above coupling reactions, a HOBt-DIC coupling agent was used. 20% piperidine in DMF was used for the Fmoc protecting group removal reaction in each step, and TFA, TIS, Thioanisole, EDT and water were mixed at a ratio of 90:2.5:2.5:2.5:2.5 in the final step of removing the resin and all protecting groups. A cocktail prepared by the above was used.

After the reaction in which each amino acid was introduced, a Kaiser test was conducted, and if there was no color change, the coupling reaction was judged to be complete and the next step reaction proceeded. The ring reaction proceeded again.

The peptide prepared in the above process was purified using purification high-performance liquid chromatography (Prep-HPLC, column C18, 10 um, 250 mm × 22 mm), and lyophilized to obtain compound 1 (LC mass molecular weight: 1097.15, yield : 85%).

[Example 2] Preparation of Compound 2

Example 2 (Compound 2) was synthesized according to solid-phase peptide synthesis. Specifically, an amine resin was used, and for each peptide, Lys, Lys, Phe, Leu, Lys, Trp, and Lys, which are amino acids using Fmoc (9-fluorenylmethoxycarbonyl) as an N-terminal protecting group, are used in the coupling reaction in order, and then Palmitic acid was used for the coupling reaction. For each of the above coupling reactions, a HOBt-DIC coupling agent was used. 20% piperidine in DMF was used for the Fmoc protecting group removal reaction in each step, and TFA, TIS, Thioanisole, EDT and water were mixed at a ratio of 90:2.5:2.5:2.5:2.5 in the final step of removing the resin and all protecting groups. A cocktail prepared by the above was used.

After the reaction in which each amino acid was introduced, a Kaiser test was conducted, and if there was no color change, the coupling reaction was judged to be complete and the next step reaction proceeded. The ring reaction proceeded again.

The peptide prepared in the above process was purified using purification high-performance liquid chromatography (Prep-HPLC, column C18, 10 um, 250 mm × 22 mm), and lyophilized to obtain compound 1 (LC mass molecular weight: 858.75, yield : 92%).

[Comparative Example 1] Preparation of Compound 3

Synthesis of Comparative Example 1 (Compound 3) was performed according to solid-phase peptide synthesis. Specifically, an amine resin was used, and for each peptide, amino acids Arg, Lys, Arg, and Lys using Fmoc (9-fluorenylmethoxycarbonyl) as an N-terminal protecting group were used in the coupling reaction in order, and each amino acid coupling reaction HOBt-DIC coupling agent was used. 20% piperidine in DMF was used for the Fmoc protecting group removal reaction in each step, and TFA, TIS, Thioanisole, EDT and water were mixed at a ratio of 90:2.5:2.5:2.5:2.5 in the final step of removing the resin and all protecting groups. A cocktail prepared by the above was used.

After the reaction in which each amino acid was introduced, a Kaiser test was conducted, and if there was no color change, the coupling reaction was judged to be complete and the next step reaction proceeded. The ring reaction proceeded again.

The peptide prepared in the above process was purified using purification high-performance liquid chromatography (Prep-HPLC, column C18, 10 um, 250 mm × 22 mm), and lyophilized to obtain compound 1 (LC mass molecular weight: 858.75, yield : 92%).

[Comparative Example 2] Preparation of compound 4

Synthesis of Comparative Example 2 (Compound 4) was performed according to solid-phase peptide synthesis. Specifically, an amine resin was used, and for each peptide, Lys, Lys, Phe, Leu, Lys, Trp, and Lys, which are amino acids using Fmoc (9-fluorenylmethoxycarbonyl) as an N-terminal protecting group, were used in the coupling reaction in order, For each amino acid coupling reaction, a HOBt-DIC coupling agent was used. 20% piperidine in DMF was used for the Fmoc protecting group removal reaction in each step, and TFA, TIS, Thioanisole, EDT and water were mixed at a ratio of 90:2.5:2.5:2.5:2.5 in the final step of removing the resin and all protecting groups. A cocktail prepared by the above was used.

After the reaction in which each amino acid was introduced, a Kaiser test was conducted, and if there was no color change, the coupling reaction was judged to be complete and the next step reaction proceeded. The ring reaction proceeded again.

The peptide prepared in the above process was purified using purification high-performance liquid chromatography (Prep-HPLC, column C18, 10 um, 250 mm × 22 mm), and lyophilized to obtain compound 4 (LC mass molecular weight: 977.25, yield : 84%).

[Experimental Example 1] Cytotoxicity (MTT assay) evaluation

HaCaT cells were seeded in a 96 well plate at a concentration of 2 x 10 ⁵ /mL, and after 24 hours, the cells were replaced with serum-free DMEM, and then treated with compounds 1 to 4 of Examples 1 to 4. After incubating the plate at 37 °C for 24 hours, the media was removed and 20 μL of 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT, 5 mg/mL) was added. was added and incubated for 2 hours at 37 °C in a CO ₂ incubator. After dissolving the crystals with 100 μL of DMSO, absorbance was measured at 570 nm. Table 1 below shows the results of cytotoxicity evaluation. Survival rate was expressed as % compared to control.

control group experimental group Concentration (ug/ml) 0 One 10 100 1,000 Example 1 100 99.3 102.4 98.5 94.4 Example 2 100 100.5 98.2 95.8 91.2 Melittin 100 92.2 52.5 23.4 14.2

As shown in Table 1, it can be seen that melittin has very high cytotoxicity with a cell viability of 52.5% even at 10ug/ml. On the other hand, in the example of the present invention, when 100 ppm, Example 1 (Compound 1) showed a high survival rate of 98.5% and Example 2 (Compound 2) 95.8%, indicating low cytotoxicity.

[Experimental Example 2] Clear zone antibacterial efficacy evaluation

For C. acnes, Blood-Agar medium (tryptic soy broth 3%, yeast extract 0.5%, 0.025% resazurin 2 mL, agar 1.5%, cysteine HCl-H2O 0.05%, hemin solution 0.5 mg/mL, vitamin K1 2 mg/mL , sheep blood 5.0%) (BRL) using an anaerobic bacterial incubator (Bactron 300: Shell Lab, IL, USA) under anaerobic conditions (anaerobic gas mixture: 85% N ₂ , 10% H ₂ , 5% CO ₂ ) was cultured in

The antibacterial activity of the peptide was performed by the paper disc method. After smearing the number of activated bacteria per plate medium to about 10 ⁷ cells, put a sterilized filter paper disc (Tokyo, 8 mm, Japan) on the solid plate medium, and then absorb the sample by concentration so that it becomes 0.05 mL / disc After culturing at 37 ° C for 24 hours, the diameter (mm) of the clear zone around the disc was measured, and the results are shown in Table 2 below.

strains Concentration (μg/mL) Clear zone(mm) Example 1 Example 2 Comparative Example 1 Comparative Example 2 Melittin C. acnes 0 - - - - - 10 15.2 13.4 - - 14.5 30 19.3 17.8 - - 18.6 100 24.6 21.5 - - 22.8 300 29.4 26.8 9.4 9.1 27.1

As shown in Table 2, the clear zone for each concentration of the peptide strain was measured, and it can be seen that the embodiment of the present invention inhibits the strain in a concentration-dependent manner.

In addition, it can be seen that the examples of the present invention exhibit significantly improved antibacterial effect against acne bacillus compared to comparative examples having very low antibacterial activity, and are equivalent to melittin, a natural antibacterial substance known to exhibit a strong antibacterial effect. By showing the level of antibacterial effect, it can be seen that the embodiment of the present invention can be used as an excellent antibacterial effective material.

[Experimental Example 3] Measurement of Minimal Inhibitory Concentration (MIC)

Antibacterial activity was measured for the strains listed in Table 3 in order to compare the antibacterial activity of Compounds 1 to 4 and Melittin (Sigma-Aldrich), which are antimicrobial peptides of Examples 1 to 4 of the present invention.

For the antibacterial activity, the minimum growth inhibitory concentration (MIC) of the peptide that does not divide the cells was measured in MH (mueller Hinton) medium with sufficient nutrients, and the strains listed in Table 1 below were obtained, so that the number of bacteria was 2X10 ⁶ CFUs per mL. After diluting with MH medium and dispensing 100 μL each to a 96 well plate, 100 μL of the sample solution diluted with MH medium was added to each well. After culturing the plate at 37 ° C. for 16 hours, the absorbance was measured at 620 nm to determine the MIC, which is shown in Table 3.

strain MIC (μg/mL) Example 1 Example 2 Comparative Example 1 Comparative Example 2 Melittin C. acnes 8 8 >128 >128 8 S. epidermidis 4 8 >128 >128 8 S. aureus 8 8 >128 >128 8 E. coli 4 8 >128 >128 8 P. aeruginosa 16 16 >128 >128 16

As shown in Table 3, it can be seen that Examples 1 and 2 of the present invention exhibit very good antibacterial activity, whereas Comparative Example has very low antibacterial activity. In addition, the examples show an equivalent level of effect compared to melittin, which is an excellent natural antibacterial material, so it can be seen that the examples of the present invention exhibit a strong antibacterial effect.

As described above, the present invention has been described by specific details, limited examples and comparative examples, but these are only provided to help a more general understanding of the present invention, and the present invention is not limited to the above examples, and the present invention Those skilled in the art can make various modifications and variations from these descriptions.

Therefore, the spirit of the present invention should not be limited to the described embodiments, and it will be said that not only the claims to be described later, but also all modifications equivalent or equivalent to these claims belong to the scope of the present invention. .

Claims (6)

A cosmetic composition for improving acne containing an antibacterial peptide derivative represented by Formula 3 as an active ingredient.
[Formula 3]

[In Formula 3,
A ₁ is a C12-C24 fatty acid.] According to claim 1,
In Formula 3, A ₁ is palmitoyl, a cosmetic composition for improving acne. delete delete delete According to claim 1,
The cosmetic composition for improving acne is a cosmetic composition for improving acne, in the form of lotion, toner, lotion, nutrient lotion, nutrient cream, massage cream, essence, pack or microneedle patch.