KR102725513B1 - Insect-derived Proteatiamycine-9 peptide with anti-inflammation and a use of the peptide - Google Patents

Abstract

The present invention relates to a novel anti-inflammatory peptide derived from white-spotted flower chafer and its use, and more particularly, to protetiamycin-9 derived from white-spotted flower chafer and an anti-inflammatory composition comprising the same. Protetiamycin-9 according to the present invention has an excellent anti-inflammatory effect and is non-cytotoxic, and thus can be utilized in various fields such as foods, pharmaceuticals, and cosmetics.

Description

{Insect-derived Proteatiamycine-9 peptide with anti-inflammation and a use of the peptide}

The present invention relates to a novel anti-inflammatory peptide derived from white-spotted flower chafer and its use, and more particularly, to protetiamycin-9 derived from white-spotted flower chafer and an anti-inflammatory composition comprising the same.

Inflammation is a defense response of living tissue to a certain stimulus, and refers to a complex lesion that causes three things: tissue degeneration, circulatory disorder, exudation, and tissue proliferation. The causes include physical factors such as mechanical injury, temperature, and radiation, chemical factors such as toxins, and parasites such as bacterial infection, and among these, bacteria are the most common. In addition to these main causes, its occurrence is complicated by various secondary factors and individual predisposition or immunity.

In addition, the inflammatory response can be said to be one of the most important responses among the pathological mechanisms according to the host's defense mechanism against wounds, microbial infections, etc. Macrophages are the most representative immune cells that regulate this inflammatory response, and activated macrophages secrete various inflammatory mediators such as TNF-α (tumor necrosis factor-α), IL-6 (interleukin-6), PGE2 (prostaglandin E2), NO (nitric oxide), and ROS (reactive oxygen species) (Laskin, D. L. et al., 2011). Meanwhile, excessive expression of these inflammatory mediators induces rheumatoid arthritis, osteoporosis, sepsis, vascular disease, cancer, etc.

Meanwhile, insects have evolved to acquire their own special physiological regulatory mechanisms and substances in order to adapt to and survive in their living environments. Recently, research is being actively conducted to develop new drug candidates using insect physiologically active substances [ Insect Biochem. Mol. Biol. 41, 747-769].

The present inventors, while continuing their research on insect-derived anti-inflammatory peptides, completed the present invention by confirming that protethiamycin-9, a novel peptide derived from white-spotted chafer, has an excellent anti-inflammatory effect.

Accordingly, it is an object of the present invention to provide novel anti-inflammatory peptides and uses thereof.

To achieve the above purpose, the present invention provides an anti-inflammatory peptide represented by the amino acid sequence of sequence number 1.

In addition, the present invention provides a pharmaceutical composition for preventing or treating inflammatory diseases, comprising the peptide as an active ingredient.

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In addition, the present invention provides an anti-inflammatory cosmetic composition containing the peptide as an effective ingredient.

In addition, the present invention provides an anti-inflammatory feed additive containing the peptide as an effective ingredient.

In addition, the present invention provides an anti-inflammatory food composition containing the peptide as an effective ingredient.

In addition, the present invention provides an anti-inflammatory health functional food composition containing the peptide as an effective ingredient.

Protetiamycin-9, a novel peptide isolated from white-spotted chafer according to the present invention, has excellent anti-inflammatory effects and no cytotoxicity, and can be utilized in various fields such as food, pharmaceuticals, and cosmetics.

Figure 1 is a diagram showing the results of confirming the cytotoxicity of proteolytic enzyme-9 according to the present invention.
Figure 2 is a diagram showing the results of confirming the anti-inflammatory activity of protetiamicin-9 according to the present invention through measurement of nitric oxide secretion.
Figure 3 is a diagram showing the results of confirming the cytokine inhibitory activity of proteolytic amine-9 according to the present invention through qRT-PCR.
Figure 4 is a diagram showing the results of confirming the inflammatory mediator inhibitory activity of proteolytic amine-9 according to the present invention through qRT-PCR.
Figure 5 is a diagram showing the results of confirming the cytokine inhibitory activity of protetiamicin-9 according to the present invention at the protein level through ELISA.

Hereinafter, the present invention will be described in detail.

The present invention provides an anti-inflammatory peptide represented by the amino acid sequence of SEQ ID NO: 1. The SEQ ID NO: 1 is represented as follows.

CVLKKAYFLTNLKLRG

In the present invention, the term "peptide" means a linear molecule formed by amino acid residues being linked to each other by peptide bonds. The peptide can be prepared by a chemical synthesis method known in the art, and preferably can be prepared by a solid phase synthesis technique, but is not limited thereto. The anti-inflammatory peptide according to the present invention can be isolated from white-spotted flower chafer.

In the present invention, the "white-spotted flower chafer ( Protaetia brevitarsis seulensis )" is an insect of the flower chafer family of the order Coleoptera, with a body length of 17 to 22 mm, and a glossy green-brown, copper-colored, or reddish color, but there are also variations such as green, blue, dark blue, and black purple. The hairs on the inside of the posterior tibial segment are gray-brown. The hairs on the inside of the posterior tibial segment are gray-brown, the tip of the mesothorax is shaped like a ginkgo, and the tip of the elytra is pointed and protrudes. The abdominal shield of the male is notched along its length. The burrow is thin and straight, and the medial plate is narrow and long with a rounded protruding part at the end.

It is preferable that the peptide according to the present invention inhibits the production of nitric oxide.

It is preferable that the peptide of the present invention inhibits the expression of cytokines and inflammatory mediators, more preferably, it inhibits the expression of cytokines IL-6 or IL-1β; and inflammatory mediators iNos or Cox-2;

The peptide according to the present invention comprises a functional equivalent of an amino acid in the amino acid sequence consisting of SEQ ID NO: 1, as long as the anti-inflammatory activity is not reduced.

The above "functional equivalent" refers to a peptide having at least 80%, preferably 90%, and more preferably 95% sequence homology (i.e., identity) with the peptide of SEQ ID NO: 1 as a result of addition, substitution, or deletion of amino acids, for example, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and 100% sequence homology, and exhibits substantially the same physiological activity as the peptide of SEQ ID NO: 1. In this specification, sequence homology and identity are defined as the percentage of amino acid residues of the candidate sequence relative to the amino acid sequence of SEQ ID NO: 1 after aligning the candidate sequence with the amino acid sequence of SEQ ID NO: 1 and introducing gaps. Where necessary, conservative substitutions are not considered as part of the sequence identity in order to obtain the maximum percentage sequence identity. N-terminal, C-terminal or internal extensions, deletions or insertions of the amino acid sequence of SEQ ID NO: 1 are not construed as sequences affecting sequence identity or homology.

The sequence identity can also be determined by standard methods commonly used to compare similar portions of the amino acid sequences of two polypeptides. Computer programs such as BLAST or FASTA align two polypeptides so that each amino acid matches optimally (along the full length of one or both sequences or along a predicted portion of one or both sequences). The programs provide a default opening penalty and a default gap penalty, and provide a scoring matrix such as PAM250 (a standard scoring matrix) that can be used in conjunction with the computer program. For example, the percent identity can be calculated as follows: the total number of identical matches multiplied by 100, then divided by the sum of the length of the longer sequence within the matched span and the number of gaps introduced into the longer sequence to align the two sequences.

As mentioned above, "substantially identical physiological activity" refers to anti-inflammation. The scope of "functional equivalents" of the present invention includes derivatives in which some of the chemical structures of the peptide are modified while maintaining the basic framework and anti-inflammatory activity of the peptide of SEQ ID NO: 1. For example, structural modifications to change the stability, storability, volatility, or solubility of the peptide are included.

In one embodiment of the present invention, a novel peptide was isolated from the white-spotted flower chafer, named protetiamycin-9, and it was confirmed that the peptide had excellent anti-inflammatory activity.

In one aspect, the present invention provides an anti-inflammatory composition comprising the peptide as an effective ingredient. The anti-inflammatory composition may be a pharmaceutical composition for preventing or treating an inflammatory disease, and may be an anti-inflammatory cosmetic composition, a pharmaceutical additive, a feed additive, a food composition, or a health functional food composition.

In addition, the anti-inflammatory composition of the present invention may contain one or more known effective ingredients having an anti-inflammatory effect together with the peptide.

The pharmaceutical composition for preventing or treating an inflammatory disease of the present invention can be manufactured by including, in addition to the effective ingredients described above, at least one pharmaceutically acceptable carrier for administration. The pharmaceutically acceptable carrier may be saline solution, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, and a mixture of at least one of these ingredients, and may further include other conventional additives such as antioxidants, buffers, and bacteriostatic agents, as needed. In addition, a diluent, a dispersant, a surfactant, a binder, and a lubricant may be additionally added to formulate the composition into an injectable formulation such as an aqueous solution, a suspension, an emulsion, a pill, a capsule, a granule, or a tablet. Furthermore, the composition may be preferably formulated according to each disease or ingredient by using an appropriate method of sugar distribution or a method disclosed in Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA.

The pharmaceutical composition for preventing or treating an inflammatory disease of the present invention can be administered orally or parenterally depending on the intended method, and the dosage range varies depending on the subject's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of the disease. The daily dosage of the peptide is 2 to 10 mg/kg, but may be increased or decreased depending on clinical results, and it is preferable to administer once or several times a day in divided doses.

The above inflammatory disease is preferably selected from the group consisting of inflammatory bowel disease, inflammatory collagen vascular disease, glomerulonephritis, inflammatory skin disease, sarcoidosis, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, pharyngitis, bronchitis, pneumonia, pancreatitis, sepsis, and nephritis, but the scope of the present invention is not limited thereto.

In another aspect, the present invention provides an anti-inflammatory cosmetic composition comprising the peptide as an effective ingredient.

The above cosmetic composition can be prepared in the form of a general emulsified formulation and a solubilized formulation. Cosmetics in the form of an emulsified formulation include a nourishing toner, a cream, an essence, etc., and cosmetics in the form of a solubilized formulation include an elastic toner. In addition, the cosmetic composition of the present invention can be prepared in the form of an auxiliary agent that can be applied locally or systemically, which is commonly used in the field of dermatology, by containing a dermatologically acceptable medium or base in addition to the above peptide. In addition, suitable cosmetic formulations can be provided in the form of a solution, gel, solid or paste anhydrous product to which the above peptide is added, an emulsion obtained by dispersing an oil phase in an aqueous phase, a suspension, a microemulsion, a microcapsule, microgranules, or an ionic (liposome) or nonionic vesicle dispersion, a cream, a skin, a lotion, a powder, an ointment, a spray, or a concealer. In addition, it can be prepared in the form of a foam or an aerosol composition further containing a compressed propellant. In addition, the cosmetic composition may further contain arbutin, kojic acid, rucinol, vitamin C, and vitamin C derivatives as excipients, and for the purpose of improving wrinkles, retinol and its derivatives, indol acetic acid and its derivatives, adenosine, tocopherol and its derivatives, kainitin, etc. In addition, the excipients may include components commonly used in cosmetic compositions, and include, for example, common auxiliary agents and carriers such as stabilizers, solubilizers, vitamins, pigments, and fragrances.

It is preferable that the anti-inflammatory cosmetic composition of the present invention contain 0.01 to 1 wt% of the peptide based on the total weight of the composition, but is not limited thereto.

In another aspect, the present invention provides an anti-inflammatory feed additive comprising the peptide as an effective ingredient.

The feed additive of the present invention is provided as a feed by being appropriately mixed with feed raw materials. As the feed raw materials, grain oil, starch, vegetable oil cake, animal feed raw materials, other feed raw materials, refined products, etc. are used, but are not limited thereto. In addition to the peptide of the present invention, the feed may additionally contain conventional additives that can increase the preservability of the feed.

The feed additive of the present invention may further include other non-pathogenic microorganisms. The microorganisms that may be added may be selected from the group consisting of Bacillus subtilis , which can produce protein-decomposing enzymes, lipolytic enzymes, and sugar-converting enzymes, Lactobacillus sp. , which has physiological activity and organic matter-decomposing ability under anaerobic conditions such as the stomach of a cow, Aspergillus oryzae , which shows effects of increasing the weight of livestock, increasing milk production, and increasing the digestibility and absorption rate of feed, and yeasts such as Saccharomyces cerevisiae .

The feed additive of the present invention may include a binder, an emulsifier, a preservative, etc. added to prevent quality deterioration, and may include an amino acid agent, a vitamin agent, an enzyme agent, a probiotic agent, a flavoring agent, a non-protein nitrogen compound, a silicate agent, a buffer agent, a coloring agent, an extractant, an oligosaccharide, etc. added to the feed to increase utility, and may additionally include a feed mixer, etc.

In another aspect, the present invention provides an anti-inflammatory food composition or health functional food composition containing the peptide as an effective ingredient.

When the composition of the present invention is used as a food composition, the food composition of the present invention means a food having a preventive or ameliorating effect on a disease caused by inflammation, and must be harmless to the human body when taken over a long period of time.

There is no particular limitation on the type of the composition. Examples of foods to which the substance can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and include all health foods in the conventional sense.

In an embodiment of the present invention, the food composition of the present invention may be a food additive. The food additive may be used by adding the peptide represented by the amino acid sequence of the above sequence number 1 as it is or together with other foods or food ingredients, and may be used appropriately according to a conventional method. The mixing amount of the effective ingredients may be appropriately determined according to the purpose of use (prevention, health, or therapeutic treatment).

In an embodiment of the present invention, the food composition of the present invention may be a health beverage composition. The health beverage composition may contain various flavoring agents or natural carbohydrates as additional ingredients, in addition to the peptide represented by the amino acid sequence of SEQ ID NO: 1, like a typical beverage. The natural carbohydrates described above may include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, or synthetic sweeteners such as saccharin and aspartame. The proportion of the natural carbohydrate is generally about 0.01 to 10 g, preferably about 0.01 to 0.1 g, per 100 ml of the composition of the present invention.

In addition to the above, the composition of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the composition of the present invention may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These components may be used independently or in combination. The proportion of these additives is not particularly important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, the contents of the present invention will be described more specifically through examples and experimental examples. The following examples and experimental examples are only intended to illustrate the present invention, and the contents of the present invention are not limited by the following examples and experimental examples.

Example 1. Selection of anti-inflammatory peptides derived from white-spotted chafer

To select peptides exhibiting anti-inflammatory activity from the white-spotted flower chafer, the following experiments were performed.

First, the intestines of the white-spotted flower chafer larvae were extracted, and the tissues except the intestines were rapidly frozen in liquid nitrogen to isolate the entire gDNA, and the genetic information on the white-spotted flower chafer genome was confirmed using an in-silico analysis method. Using the previously known anti-inflammatory peptide properties, the entire sequence of the translated amino acids from each gene was filtered, and peptides predicted to be new anti-inflammatory peptides were selected.

The selected amino acid sequence was CVLKKAYFLTNLKLRG, consisting of 16 amino acids, and was named Protaetiamycine-9 (Table 1).

Peptide Amino acid sequence Protetiamicin-9
(sequence number 1) CVLKKAYFLTNLKLRG-NH ₂

Example 2. Synthesis and purification of protetiamicin-9

Protetiamycin-9 peptide derived from the white-spotted flower chafer selected in Example 1 above was synthesized, isolated, and purified.

For this purpose, the peptide was prepared using Merrifield's liquid-solid method (Merrifield, RB., J. Am. Chem. Soc., 85, 2149, 1963) using an Fmoc amino group protecting vessel. The peptide was synthesized by a solid phase method using Fmoc (9-fluorenylmethoxycarbonyl) as a protecting group for the Nα-amino group of the amino acid.

Specifically, peptides having a carboxyl terminal in the -NH2 form were used as a starting material from Rink Amide MBHA-Resin, and peptides having a carboxyl terminal in the -OH form were used as a starting material from Fmoc-amino acid-Wang Resin. The elongation of the peptide chain by coupling of Fmoc-amino acid was performed by the DCC (N-hydroxybenzo-triazole (HOBt)-dicyclo-hexylcarboxyl-bodiimide) method. After coupling the Fmoc-amino acid at the amino terminus of each peptide, the Fmoc group was removed with a 20% piperidine/N-methylpyrrolidone (NMP) solution, washed several times with NMP and dichloromethane (DCM), and then dried with nitrogen gas. Here, a solution of trifluoroacetic acid (TFA)-phenol-thioanisole-H2O-triisop- ropylsilane (85: 5: 5: 2.5: 2.5, vol./vol.) was added and reacted for 3 h to remove the protecting group and separate the peptide from the resin, which was then precipitated with diethyl ether. The crude peptide thus obtained was purified using a reverse phase-HPLC column (Delta Pak, C18 300Å, 15μ, 19.0 mm×30, Waters) with an acetonitrile gradient containing 0.1% TFA. The synthetic peptide was hydrolyzed with 6 N-HCl at 110°C for 24 hours, the resulting residue was concentrated under reduced pressure, dissolved in 0.02 N-HCl, and the amino acid composition was measured using an amino acid analyzer (Hitachi 8500 A).

Additionally, the molecular weight was calculated based on the sequence of the synthesized peptide, and the exact molecular weight was measured using matrix-assisted laser desorption ionization mass spectrometer (MALDI).

As a result, it was confirmed that protetiamicin-9 having the correct amino acid sequence was synthesized because the measured molecular weight and the calculated molecular weight matched. This was used in subsequent experiments.

Experimental Example 1. Confirmation of cytotoxicity of protetiamicin 9

1-1. Cell culture

To verify the anti-inflammatory effect, experiments were conducted using Raw264.7 cells, which are mouse macrophages. They were cultured in DMEM medium containing antibiotics (100 units/ml penicillin, 100 μg/ml streptomycin) and 10% fetal bovine serum (FBS) in a 5% CO2 incubator at ₃₇ °C and used in the experiment.

1-2. MTS assay

2 × 10 ⁴ cells/well were seeded in 200 μl in a 96-well culture dish and treated with protetiamycin 9 at various concentrations for 24 hours. After that, 20 μl of MTS reagent CellTiter 96 ^ⓡ AQueous One Solution Reagent (Promega, USA) was added and the reaction was performed for 3 to 4 hours to check the color change, which was confirmed by measuring the absorbance at a wavelength of 450 nm using a multi detector (Beckman DTX8800). The results of checking the cell viability are shown in Fig. 1.

As shown in Figure 1, the proteolytic acid-9 peptide derived from the white-spotted chafer was confirmed to have almost no cytotoxicity up to a concentration of 200 ug/ml.

Experimental Example 2. Anti-inflammatory activity assay of protetiamicin 9

2-1. Nitric oxide (NO) assay

To measure the anti-inflammatory activity of proteolytic enzyme 9, the amount of NO secretion in Raw264.7 cells was confirmed. After preparing the cells in 6-well at a cell number of 1 × 10 ⁶ cells/ml, the peptides were treated at various concentrations for 1 hour. Then, 100 ng/ml of lipopolysaccharide (LPS) was treated to induce inflammation and cultured for 24 hours. 100 μl of the supernatant of the cultured cells was used in the experiment, and the amount of NO secreted by LPS was confirmed to be reduced by the peptide using a NO detection kit (intron) by measuring the absorbance at a wavelength of 550 nm using a multi detector (Beckman DTX8800). The results of confirming the amount of NO secretion are shown in Fig. 2.

As shown in Figure 2, it was confirmed that the protetiamicin-9 peptide derived from the white-spotted flower chafer decreased the amount of NO secretion in a concentration-dependent manner.

2-2. Quantitative RT-PCR (qRT-PCR)

The expression of cytokines (IL-6, IL-1β); and inflammatory mediators (iNos, Cox-2) was confirmed through qRT-PCR. After preparing cells in 6-well at a cell number of 1 × 10 ⁶ cells/ml, proteolytic 9 peptide was treated at various concentrations for 1 hour. After that, 100 ng/ml of LPS was treated to induce inflammation and incubated for 18 hours. After that, the supernatant was removed, washed with PBS, and the cells were harvested. 1 ml of TRIZOL solution was mixed with the harvested cells and reacted at room temperature for 5 minutes. After adding 200 μl chloroform and mixing, centrifugation was performed at 12,000×g for 15 minutes at 4℃. The supernatant was collected, 500 μl isopropanol was added, reaction was performed at room temperature for 10 minutes, and centrifugation was performed at 12,000×g for 10 minutes at 4℃. The precipitated RNA was washed with 1 ml 75% ethanol and centrifuged at 7,500×g for 5 minutes at 4℃. After centrifugation, ethanol was removed, dried, and then dissolved in sterilized water to prepare RNA. After that, cDNA was prepared using a cDNA synthesis kit using 1 μg of total RNA, and qRT-PCR was performed. The primers used for analysis of cytokine and inflammatory mediator expression are as shown in Table 2, and the experimental results are shown in Figures 3 and 4, respectively.

Name Sequence IL-6 Forward 5’-GAGGATACCACTCCCAACAGACC-3’ Reverse 5’-AAGTGCATCATCGTTGTTCATACA-3’ IL-1β Forward 5’-CCTTCCAGGATGAGGACATGA-3’ Reverse 5’-TGAGTCACAGAGGATGGGCTC-3’ i-NOS Forward 5’-CAGCACAGGAAATGTTTTCAGC-3’ Reverse 5’-TAGCCAGCGTACGGATGA-3’ COX-2 Forward 5’-CAGACAACATAAACTGCGCCTT-3’ Reverse 5’-GATACACCTCTCCACCAATGACC-3’ GAPDH Forward 5’-AAGGTCATCCCAGAGCTGAA-3’ Reverse 5’-CTGCTTCACCACCTTCTTGA-3’

As shown in Figures 3 and 4, it was confirmed that the proteolytic peptide derived from white-spotted flower chafer inhibited the expression of cytokines (IL-6, IL-1β); and inflammatory mediators (iNos, Cox-2) in a concentration-dependent manner.

2-3. Enzyme-linked immunosorbent assay (ELISA)

Raw264.7 cells were prepared in 6-well at a cell number of 1 × 10 ⁶ cells/ml each, and then treated with protetiamicin 9 peptide at various concentrations for 1 hour. Then, 100 ng/ml of LPS was treated to induce inflammation and cultured for 24 hours. The supernatant of the cultured cells was collected, and the secreted amounts of TNF-α, IL-6, and IL-1β were quantified by measuring the absorbance at a wavelength of 450 nm with a multi detector (Beckman DTX8800) using an ELISA kit. The quantitative results are shown in Fig. 5.

As shown in Figure 5, it was confirmed that the protetiamicin-9 peptide derived from white-spotted flower chafer inhibited the expression of cytokines IL-6 and IL-1β in a concentration-dependent manner at the protein level.

In summary, protetiamycin-9, a novel peptide isolated by the present inventors from white-spotted chafer, has excellent anti-inflammatory effects and no cytotoxicity, and can be utilized in various fields such as food, pharmaceuticals, and cosmetics.

<110> REPUBLIC OF KOREA(MANAGEMENT: RURAL DEVELOPMENT ADMINISTRATION) <120> Insect-derived Proteatiamycine-9 peptide with anti-inflammation and a use of the peptide <130> 1-197 <160> 1 <170> KoPatentIn 3.0 <210 > 1 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> Protaetia brevitarsis <400> 1 Cys Val Leu Lys Lys Ala Tyr Phe Leu Thr Asn Leu Lys Leu Arg Gly 1 5 10 15

Claims (10)

An anti-inflammatory peptide represented by the amino acid sequence of sequence number 1. In the first paragraph, the peptide is a peptide isolated from white-spotted flower chafer. In claim 1, the peptide is a peptide that inhibits the production of nitric oxide. In claim 1, the peptide is a peptide that inhibits the expression of IL-6, IL-1β, iNos or Cox-2. delete delete An anti-inflammatory cosmetic composition comprising a peptide according to any one of claims 1 to 4 as an active ingredient. An anti-inflammatory feed additive comprising a peptide according to any one of claims 1 to 4 as an active ingredient. An anti-inflammatory food composition comprising a peptide according to any one of claims 1 to 4 as an effective ingredient. An anti-inflammatory health functional food composition comprising a peptide according to any one of claims 1 to 4 as an effective ingredient.