Novel anti-inflammatory peptides and their uses

Abstract

The present invention relates to a novel immunoregulatory peptide having an inflammatory response control function and its use. The peptide of the present invention can be usefully used in the treatment of inflammation, various immune diseases, and cancer by regulating the activity of immune cells.

Description

[Technical field]

[0001] The present invention relates to a novel peptide, a pharmaceutical composition for preventing or treating immune diseases or cancer, and an anti-inflammatory cosmetic composition containing the same. [Background technology]

[0002] Resolution of inflammation is a biological mechanism that ends the innate immune response and restores tissue homeostasis and induces an appropriate adaptive immune response. In the past, the resolution of inflammation was understood to be a passive phenomenon that occurs after an inflammatory response, but recently, as mediators of inflammation resolution have been identified, it has been revealed that it is an active biological mechanism that progresses stepwise through an inflammatory response with the involvement of multiple factors. When an injury, contact with an external substance, or infection occurs, a defense action occurs through the acute inflammatory response of the innate immune system, and then the resolution of inflammation is activated to control the inflammatory response to an appropriate level and induce healing and recovery of the damaged tissue. The resolution of inflammation restricts further infiltration of neutrophils into the inflamed tissue, induces cell death, and mediates the reduction of lipid inflammatory substances, cytokines, and chemokines that cause an inflammatory response. It then promotes the removal of damaged tissue or dead cells by inducing efferocytosis in macrophages, thereby reducing inflammatory macrophages and increasing macrophages with anti-inflammatory properties. Appropriate inflammation resolution mediates normal adaptive immune responses and strengthens the ability to respond to future repeated inflammatory situations, but insufficient inflammation resolution can lead to transient / persistent inflammatory responses, inducing various chronic inflammatory diseases, and abnormal adaptive immune responses can cause autoimmune diseases.

[0003] The inflammation-resolving response is induced through cell-secreted pro-resolving factors, also known as specialized pro-resolving mediators (SPMs). When the inflammation-resolving response begins, various forms of inflammation-resolving factors, such as lipids, peptides, and proteins, control the inflammatory response, including the activation of immune cells, through specific receptors such as GPCRs.

[0004] Currently, anti-inflammatory drugs suppress the occurrence of inflammatory reactions by suppressing the activity of specific enzymes or antagonizing specific receptors or their ligands. Existing anti-inflammatory strategies can effectively and strongly suppress acute inflammatory reactions, but they also suppress normal immune responses, resulting in various side effects. In particular, there are safety concerns such as a weakened immune system and the risk of secondary infection in patients with chronic inflammation / autoimmune diseases who require long-term drug administration. Therefore, there is an emerging need to develop a safe therapeutic agent with efficacy and safety in controlling inflammatory reactions through a new approach. The inflammation resolution induction strategy is an innovative immune treatment strategy that uses agonists that enhance the mechanism of inflammation resolution to control transient / sustained inflammatory reactions. In particular, it is attracting attention as an alternative to broad-spectrum immunosuppressants, which have a high risk of side effects, in the treatment of chronic inflammation / autoimmune diseases that require long-term drug administration due to their limited effect on normal immune responses. The targets of the therapeutic strategy of inducing inflammation resolution include inflammatory diseases caused by immune abnormalities in various tissues such as the skin, respiratory system, digestive system, eyeball, musculoskeletal system, and circulatory system, as well as diseases in which the inflammatory response caused by the onset of the disease has a significant impact on the progression of the disease. In particular, Parkinson's disease and Alzheimer's disease, which are neurological diseases well known to have a significant impact on the worsening of symptoms, are representative examples. In addition, the importance of the influence of inflammatory responses in the tumor microenvironment on the progression of cancer, especially in solid cancers, has been confirmed, and anti-cancer treatment through immune regulation through the induction of inflammation resolution is also being evaluated as a new strategy.

[0005] FPR (Formyl-Peptide Receptor) series receptors are G protein-coupled receptors (GPCRs) that exist mainly in immune cells and mediate various cellular functions through downstream signaling systems. FPR1 mainly binds to bacterial peptides to induce acute inflammatory responses and mediate defense, while FPR2 has been revealed to control inflammatory responses by binding to endogenous ligands of various forms (peptides, proteins, lipids), and is therefore attracting attention as a target for inflammatory disease treatments. FPR2 is a receptor for representative inflammation resolution factors (LXA4, AnxA1, Ac2-26, RvD1, etc.) that are expressed in endogenous immune cells and mediate the induction of inflammation resolution responses. Based on the characteristics of FPR2 with multiple ligands, attempts are being made to develop various forms of substances such as peptides, lipids, proteins, and low molecular weight compounds as FPR2-targeted inflammation resolution-inducing treatments. The Company's main focus is on developing therapeutic agents based on research results into the effectiveness of various inflammatory diseases, such as atopic dermatitis, psoriasis, asthma, lung injury, lung inflammation, conjunctivitis, keratitis, dry eye syndrome, rheumatoid arthritis, ankylosing spondylitis, ulcerative colitis, Crohn's disease, and systemic sclerosis, which are inflammatory diseases that appear in various tissues, including the skin, respiratory system, digestive system, eyeballs, musculoskeletal system, and circulatory system, due to abnormalities in local or systemic immune responses.In addition, the importance of neuroinflammation generated from microglial cells and nerve cells, which perform immune functions in the brain, in degenerative brain diseases such as Parkinson's disease and Alzheimer's disease, and the importance of the interaction between cancer cells and the inflammatory response generated from immune cells and basal cells in the tumor microenvironment in the progression of cancer have been confirmed, and many research and development results have been reported regarding the effectiveness of neurological diseases and cancer treatment strategies through inflammation relief, which is considered to be a new approach. In particular, clinical trials of FPR2-targeted peptides and small molecule compounds are underway for atopic dermatitis, heart failure, myocardial infarction, cardiovascular thrombosis, Crohn's disease, etc.

[0006] In previous research, the inventor has developed a peptide with antibacterial activity and secured a patent for it (Patent Document 1), and has also developed a peptide with excellent antibacterial and immune function regulating capabilities and secured a patent for it (Patent Document 2). [Prior art documents] [Patent documents]

[0007] [Patent Document 1] Republic of Korea Patent No. 10-1855170 [Patent Document 2] Republic of Korea Patent No. 10-2170236 Summary of the Invention [Problem to be solved by the invention]

[0008] An object of the present invention is to provide a novel peptide that has an immunoregulatory function and a therapeutic effect on immune diseases by inducing an anti-inflammatory response.

[0009] Another object of the present invention is to provide various uses of the peptide, such as pharmaceutical compositions for preventing or treating immune diseases or cancer, and anti-inflammatory cosmetic compositions. [Means for solving the problem]

[0010] In order to achieve the above object, the present invention provides a peptide having an amino acid sequence represented by X1-X2-X3-X4-X5-X6-m,

[0011] The peptide further includes a peptide in which X1 is absent, arginine (R), lysine (K), histidine (H), or proline (P); X2 is methionine (M) or tryptophan (W); X3 is a basic amino acid that is arginine (R) or lysine (K); X4 is tyrosine (Y); X5 is a basic amino acid that is arginine (R) or lysine (K), or is an aromatic amino acid that is tryptophan (W) or tyrosine (Y); X6 is proline (P), tyrosine (Y), or valine (V); and m is D-methionine.

[0012] The present invention also relates to a peptide having an amino acid sequence represented by X2-X3-X4-X5-X6-m,

[0013] The peptide further comprises X2 being isoleucine (I), proline (P) or histidine (H); X3 being lysine (K); X4 being tyrosine (Y) or phenylalanine (F); X5 being lysine (K) or tryptophan (W); X6 being valine (V), proline (P), tyrosine (Y) or methionine (M); and m being D-methionine.

[0014] The present invention relates to a peptide having an amino acid sequence represented by X1-X2-X3-X4-X5-X6-m,

[0015] The peptide provides a peptide in which X1 is a basic amino acid, arginine (R) or lysine (K); X2 is an aromatic amino acid, tryptophan (W); X3 is a basic amino acid, arginine (R) or lysine (K); X4 is tyrosine (Y); X5 is arginine (R), lysine (K), leucine (L), asparagine (N), phenylalanine (F) or threonine (T); X6 is isoleucine (I), threonine (T), tyrosine (Y) or valine (V); and m is D-methionine.

[0016] In one embodiment of the present invention, there is provided a pharmaceutical composition for preventing or treating immune diseases, comprising the peptide as an active ingredient.

[0017] In yet another embodiment of the present invention, there is provided a pharmaceutical composition for preventing or treating cancer, comprising the peptide as an active ingredient.

[0018] In yet another embodiment of the present invention, there is provided an anti-inflammatory cosmetic composition comprising the peptide as an active ingredient. Effect of the Invention

[0019] The novel peptide according to the present invention has a function of regulating the activity of immune cells and has a function of treating immune diseases or cancer by inducing an inflammation-resolving response, and therefore can be used as a pharmaceutical composition for treating immune diseases or cancer or an anti-inflammatory cosmetic composition containing the same. [Brief description of the drawings]

[0020] [Figure 1] FIG. 1 shows an analysis of the ability of peptides to suppress the production of inflammatory cytokines at the individual level. [Diagram 2] FIG. 2 is a graph showing the analysis of the efficacy of peptides in improving severity scores for collagen-induced arthritis. [Diagram 3] FIG. 3 shows an analysis of corneal damage (A) and corneal opacity (B) caused by peptide administration in a BAC-induced xerophthalmia model. [Figure 4]FIG. 4 shows an analysis of body weight (A) and colon length (B) after peptide administration in a DSS-induced colitis model. [Diagram 5] FIG. 5 shows the analysis of the ability of peptides to suppress cytokine expression in the SK-N-MC neuronal cell line (A) and rat DRG neurons (B). [Figure 6] FIG. 6 shows an analysis of the improvement of nerve terminal density (A) and neuronal number (B) of dopamine neurons in an MPTP-induced Parkinson's disease model by peptides. [Figure 7] FIG. 7 is a graph analyzing the tumor growth suppression efficacy of peptides. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021] The present invention will be described in detail below.

[0022] The present invention provides novel peptides that bind to the FPR2 receptor and function to regulate inflammatory responses.

[0023] In one aspect of the present invention, there is provided a peptide having an amino acid sequence represented by X1-X2-X3-X4-X5-X6-m,

[0024] The peptide further includes a peptide in which X1 is absent, arginine (R), lysine (K), histidine (H), or proline (P); X2 is methionine (M) or tryptophan (W); X3 is a basic amino acid that is arginine (R) or lysine (K); X4 is tyrosine (Y); X5 is a basic amino acid that is arginine (R) or lysine (K), or is an aromatic amino acid that is tryptophan (W) or tyrosine (Y); X6 is proline (P), tyrosine (Y), or valine (V); and m is D-methionine.

[0025] As used herein, the term "peptide" refers to a polymer composed of one or more amino acids linked by amide bonds (or peptide bonds), and includes not only peptides but also pharma- ceutically acceptable salts of peptides. Typically, the amino terminus (or N-terminus) of a peptide is shown at the left end of a written amino acid sequence of the peptide, and the carboxyl terminus (or C-terminus) is shown at the right end of the written amino acid sequence. The amino acid sequence of a peptide may be written using single-letter symbols to indicate the amino acids covalently linked by peptide bonds. If the D-configuration is not specifically indicated, the L-configuration may be assumed, and the amino acid residues constituting the peptide may be natural or non-natural amino acid residues.

[0026] The peptide of the present invention can be obtained by various methods widely known in the art, for example, by extracting proteins from living organisms and treating them with protease to reduce their molecular weight, or by biological production using gene recombination and a protein expression system, or by in vitro synthesis through chemical synthesis such as peptide synthesis, or by cell-free protein synthesis.

[0027] In one embodiment of the present invention, X1 is absent, arginine (R), lysine (K), histidine (H) or proline (P); X2 is methionine (M) or tryptophan (W); X3 is a basic amino acid that is arginine (R) or lysine (K); X4 is tyrosine (Y); X5 is a basic amino acid that is arginine (R) or lysine (K); X6 is proline (P), tyrosine (Y) or valine (V); and m may be D-methionine.

[0028] In one embodiment of the present invention, X1 is absent, arginine (R) or lysine (K); X2 is methionine (M) or tryptophan (W); X3 is a basic amino acid that is arginine (R) or lysine (K); X4 is tyrosine (Y); X5 is an aromatic amino acid that is tryptophan (W); X6 is proline (P), tyrosine (Y) or valine (V); and m may be D-methionine.

[0029] In one embodiment of the present invention, X1 is absent, arginine (R) or lysine (K); X2 is methionine (M) or tryptophan (W); X3 is arginine (R) as a basic amino acid; X4 is tyrosine (Y); X5 is tyrosine (Y) as an aromatic amino acid; X6 is proline (P), tyrosine (Y) or valine (V); and m may be D-methionine.

[0030] In one embodiment of the present invention, X1 is absent, arginine (R), lysine (K), histidine (H) or proline (P); X2 is methionine (M) or tryptophan (W); X3 is a basic amino acid that is arginine (R) or lysine (K); X4 is tyrosine (Y); X5 is a basic amino acid that is lysine (K); X6 is proline (P), tyrosine (Y) or valine (V); and m may be D-methionine.

[0031] In one embodiment of the present invention, X1 is absent or arginine (R); X2 is methionine (M) or tryptophan (W); X3 is a basic amino acid that is arginine (R) or lysine (K); X4 is tyrosine (Y); X5 is an aromatic amino acid that is tryptophan (W); X6 is proline (P), tyrosine (Y) or valine (V); and m may be D-methionine.

[0032] In one embodiment of the present invention, X1 is absent or arginine (R); X2 is methionine (M) or tryptophan (W); X3 is arginine (R) as a basic amino acid; X4 is tyrosine (Y); X5 is tyrosine (Y) as an aromatic amino acid; X6 is proline (P), tyrosine (Y) or valine (V); and m may be D-methionine.

[0033] In the present invention, one or more amino acids of the peptide may be chemically modified to obtain better chemical stability, enhanced pharmacological properties (half-life, water absorption, potency, efficacy, etc.), altered specificity (e.g., broader biological activity spectrum), and reduced antigenicity. The term "stability" is used to mean not only in vivo stability that protects the peptide of the present invention from attack by protease in the body, but also storage stability (e.g., room temperature storage stability). For example, a protecting group may be added to the N-terminus or C-terminus of the peptide of the present invention. Preferably, the protecting group may be an acetyl group, a butanoyl group, a hexanoyl group, an octanoyl group, a fluorenylmethoxycarbonyl group, a palmitoyl group, a myristoyl group, a stearyl group, a butoxycarbonyl group, an allyloxycarbonyl group, or a polyethylene glycol (PEG), but any component that can improve the modification of the peptide, particularly the stability of the peptide, may be included without limitation.

[0034] In the present invention, the N-terminal amino acid of a peptide may be alkylated or acylated at the N-terminal amino group (HN-). For example, the N-terminal amino group of a peptide may be acylated to include an aliphatic acyl group (i.e., including acetyl, myristoyl, butanoyl, etc.), a carboxylic acid, a benzoic acid, a trifluoroacetic acid, or a sulfonic acid. As another example, the N-terminal amino group of a peptide may be alkylated with an aliphatic alkyl, a halide, or an aliphatic alkynesulfonate.

[0035] In the present invention, the C-terminal carboxyl group (-COOH) of a peptide may be amidated or esterified. For example, the C-terminal carboxyl group may be chemically modified by forming an amide with an amine. As another example, the C-terminal carboxyl group may be chemically modified by forming an ester with an alcohol.

[0036] In addition, the side chains of the amino acids of the peptides of the present invention may be chemically modified. For example, the phenyl group of tyrosine may be substituted with, but is not limited to, an aliphatic alkyl group, an aliphatic carboxyl group, or an alkoxy group. The epsilon amino group of lysine may be chemically modified by, for example, amide formation with an aliphatic carboxylic acid group, a benzoic acid group, or an amino acid group. The epsilon amino group of lysine may be chemically modified by, but is not limited to, alkylation with one or two C1-C4 aliphatic alkyl groups.

[0037] In one embodiment of the present invention, the peptide may have any one selected from the group consisting of an acetyl group, a butanoyl group, a hexanoyl group, and an octanoyl group added to its N-terminus.

[0038] In one embodiment of the present invention, the C-terminus of the peptide may be amidated.

[0039] In one embodiment of the present invention, the peptide may be composed of any one of the amino acid sequences of SEQ ID NOs: 1 to 127.

[0040] In one aspect of the present invention, there is provided a peptide having an amino acid sequence represented by X2-X3-X4-X5-X6-m,

[0041] The peptide further comprises X2 being isoleucine (I), proline (P) or histidine (H); X3 being lysine (K); X4 being tyrosine (Y) or phenylalanine (F); X5 being lysine (K) or tryptophan (W); X6 being valine (V), proline (P), tyrosine (Y) or methionine (M); and m being D-methionine.

[0042] In one embodiment of the present invention, X2 is isoleucine (I), proline (P) or histidine (H); X3 is lysine (K); X4 is tyrosine (Y) or phenylalanine (F); X5 is lysine (K) or tryptophan (W); X6 is valine (V) or proline (P); and m may be D-methionine.

[0043] In one embodiment of the present invention, X2 is isoleucine (I) or histidine (H); X3 is lysine (K); X4 is tyrosine (Y) or phenylalanine (F); X5 is lysine (K) or tryptophan (W); X6 is tyrosine (Y) or methionine (M); and m may be D-methionine.

[0044] In one embodiment of the present invention, X2 is isoleucine (I) or proline (P); X3 is lysine (K); X4 is tyrosine (Y) or phenylalanine (F); X5 is lysine (K) or tryptophan (W); X6 is valine (V); and m may be D-methionine.

[0045] In one embodiment of the present invention, X2 is isoleucine (I) or histidine (H); X3 is lysine (K); X4 is tyrosine (Y) or phenylalanine (F); X5 is lysine (K) or tryptophan (W); X6 is proline (P); and m may be D-methionine.

[0046] In one embodiment of the present invention, X2 is isoleucine (I); X3 is lysine (K); X4 is phenylalanine (F); X5 is lysine (K) or tryptophan (W); X6 is tyrosine (Y) or methionine (M); and m may be D-methionine.

[0047] In one embodiment of the present invention, X2 may be isoleucine (I) or proline (P); X3 may be lysine (K); X4 may be tyrosine (Y) or phenylalanine (F); X5 may be lysine (K); X6 may be valine (V); and m may be D-methionine.

[0048] In one embodiment of the present invention, X2 is isoleucine (I) or histidine (H); X3 is lysine (K); X4 is tyrosine (Y) or phenylalanine (F); X5 is lysine (K) or tryptophan (W); X6 is proline (P); and m may be D-methionine.

[0049] In one embodiment of the present invention, X2 is isoleucine (I); X3 is lysine (K); X4 is phenylalanine (F); X5 is lysine (K) or tryptophan (W); X6 is tyrosine (Y) or methionine (M); and m may be D-methionine.

[0050] In one embodiment of the present invention, the peptide may have any one selected from the group consisting of an acetyl group, a butanoyl group, a hexanoyl group, and an octanoyl group added to its N-terminus.

[0051] In one embodiment of the present invention, the C-terminus of the peptide may be amidated.

[0052] In one embodiment of the present invention, the peptide may be composed of any one of the amino acid sequences of SEQ ID NOs: 128 to 151.

[0053] In one aspect of the present invention, there is provided a peptide having an amino acid sequence represented by X1-X2-X3-X4-X5-X6-m,

[0054] The peptide provides a peptide in which X1 is a basic amino acid, arginine (R) or lysine (K); X2 is an aromatic amino acid, tryptophan (W); X3 is a basic amino acid, arginine (R) or lysine (K); X4 is tyrosine (Y); X5 is arginine (R), lysine (K), leucine (L), asparagine (N), phenylalanine (F) or threonine (T); X6 is isoleucine (I), threonine (T), tyrosine (Y) or valine (V); and m is D-methionine.

[0055] In one embodiment of the present invention, X1 is a basic amino acid, arginine (R) or lysine (K); X2 is an aromatic amino acid, tryptophan (W); X3 is a basic amino acid, arginine (R) or lysine (K); X4 is tyrosine (Y); X5 is arginine (R) or lysine (K); X6 is isoleucine (I) or threonine (T); and m may be D-methionine.

[0056] In one embodiment of the present invention, X1 is a basic amino acid, arginine (R) or lysine (K); X2 is an aromatic amino acid, tryptophan (W); X3 is a basic amino acid, arginine (R) or lysine (K); X4 is tyrosine (Y); X5 is leucine (L), asparagine (N), phenylalanine (F) or threonine (T); X6 is tyrosine (Y) or valine (V); and m may be D-methionine.

[0057] In one embodiment of the present invention, X1 is a basic amino acid, arginine (R); X2 is an aromatic amino acid, tryptophan (W); X3 is a basic amino acid, arginine (R); X4 is tyrosine (Y); X5 is arginine (R) or lysine (K); X6 is isoleucine (I) or threonine (T); and m may be D-methionine.

[0058] In one embodiment of the present invention, X1 is a basic amino acid, arginine (R); X2 is an aromatic amino acid, tryptophan (W); X3 is a basic amino acid, arginine (R); X4 is tyrosine (Y); X5 is leucine (L), asparagine (N), phenylalanine (F) or threonine (T); X6 is tyrosine (Y) or valine (V); and m may be D-methionine.

[0059] In one embodiment of the present invention, the peptide may have any one selected from the group consisting of an acetyl group, a butanoyl group, a hexanoyl group, and an octanoyl group added to its N-terminus.

[0060] In one embodiment of the present invention, the C-terminus of the peptide may be amidated.

[0061] In one embodiment of the present invention, the peptide may be composed of any one of the amino acid sequences of SEQ ID NOs: 152 to 160.

[0062] In the present specification, the peptide can act as an immunomodulatory peptide that plays a role in immunomodulation, such as directly or indirectly regulating the inflammatory response of immune cells, migration of immune cells, and regulating the inflammatory response of epidermal or endothelial cells. In the present invention, the immunomodulatory peptide can exhibit an immunomodulatory function by inducing an inflammation resolution response through activation of formyl peptide receptor 2 (FPR2) expressed in immune cells, such as phagocytic cells, such as neutrophils and monocytes, antigen presenting cells, such as dendritic cells, eosinophils, basophils, mast cells, and microglia, as well as nerve cells, epidermal cells, endothelial cells, and the like.

[0063] As confirmed by the following experimental examples, the peptide according to the present invention exhibits high FPR2 activation efficacy even at low concentrations, and is highly selective compared to FPR1. In addition, the present inventors measured the expression changes of intracellular cytokine genes under inflammatory conditions to confirm whether the peptide according to the present invention reduces intracellular inflammatory responses, and confirmed that inflammatory cytokines (TNFα, IL-6, IL-1β) were reduced compared to the control group, thereby confirming the inflammatory response suppression efficacy of the peptide according to the present invention at the individual level in an animal model.

[0064] Therefore, the present invention provides a pharmaceutical composition for preventing or treating immune diseases comprising the peptide as an active ingredient, use of the peptide for preventing or treating immune diseases, and a method for preventing or treating immune diseases comprising administering the peptide to a subject.

[0065] In one embodiment of the present invention, the immune disease may be, but is not limited to, atopic dermatitis, psoriasis, infantile eczema, sepsis, conjunctivitis, keratitis, dry eye syndrome, asthma, lung injury, lung inflammation, rheumatoid arthritis, ankylosing spondylitis, ulcerative colitis, Crohn's disease, peritonitis, systemic sclerosis, neuroinflammation, Parkinson's disease, or Alzheimer's disease.

[0066] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating atopic dermatitis. Further details are disclosed in Baxendell et al. J Immunol May 1, 2018, 200(1 Supplement)105.4;

[0067] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating psoriasis. Further details are disclosed in Liu et al. Sci Rep. 2017 Aug 2; 7(1): 7100. doi: 10.1038 / s41598-017-07485-1.

[0068] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating infantile eczema. Further details are disclosed in Wu et al. Br J Dermatol. 2013 Jan; 168(1): 172-8. doi: 10.1111 / j.1365-2133.2012.11177.x.

[0069] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating sepsis. Further details are disclosed in Walker et al. SHOCK, Vol. 36, No. 4, pp. 410Y416, 2011.

[0070] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating conjunctivitis. Further details are disclosed in Hodges et al. Mucosal Immunol. 2017 Jan;10(1):46-57. doi:10.1038 / mi.2016.33. Epub 2016 Apr 13.

[0071] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating keratitis. Further details are disclosed in Zhu et al. Int Immunopharmacol. 2021 Jul;96:107785.doi:10.1016 / j.intimp.2021.107785.Epub 2021 May 24.

[0072] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating dry eye syndrome. Further details are disclosed in Gao et al. J Immunol. 2015 Oct 1; 195(7): 3086-99. doi: 10.4049 / jimmunol.1500610. Epub 2015 Aug 31.

[0073] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating asthma. Further details are disclosed in Barnig et al. Eur Respir Rev. 2015 Mar;24(135):141-53. doi:10.1183 / 09059180.00012514.

[0074] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating lung injury. Further details are disclosed in Ba et al. J Thorac Dis. 2019 Aug; 11 (8): 3599-3608. doi: 10.21037 / jtd.2019.08.86.

[0075] In the present invention, the immunomodulatory peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating pulmonary inflammation. Further details are disclosed in Sekheri et al. Proc Natl Acad Sci USA 2020 Apr 7; 117 (14): 7971-7980. doi: 10.1073 / pnas.1920193117. Epub 2020 Mar 23.

[0076] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating rheumatoid arthritis. Further details are disclosed in Kao et al. Br J Pharmacol. 2014 Sep; 171(17): 4087-96. doi: 10.1111 / bph.12768.

[0077] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating ankylosing spondylitis. Further details are disclosed in Kao et al. Br J Pharmacol. 2014 Sep; 171(17): 4087-96. doi: 10.1111 / bph.12768.

[0078] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating ulcerative colitis or Crohn's disease. For more detailed information, please refer to Kim et al. Exp Mol Med. 2013 Sep 13; 45(9): e40. doi: 10.1038 / emm.2013.77.

[0079] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating systemic sclerosis. Further details are disclosed in Park et al. Front Immunol. 2019 Sep 3; 10: 2095. doi: 10.3389 / fimmu.2019.02095. eCollection 2019.

[0080] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating neuroinflammation. Further details are disclosed in Ponce et al. Front Aging Neurosci. 2022 Feb 17; 14: 780811. doi: 10.3389 / fnagi.2022.780811. eCollection 2022.

[0081] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating Parkinson's disease. Further details are disclosed in Krashia et al. Nat Commun. 2019 Sep 2; 10(1): 3945. doi: 10.1038 / s41467-019-11928-w.

[0082] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating Alzheimer's disease. Further details are disclosed in Pamplona et al. Transl Psychiatry. 2022 Oct 10; 12(1): 439. doi: 10.1038 / s41398-022-02208-1.

[0083] In the present invention, the peptide can act as an anti-inflammatory factor that binds to the FPR2 receptor.

[0084] Furthermore, the present inventors have confirmed that tumor growth is suppressed when the peptide of the present invention is administered to an animal model, and therefore that the peptide that induces the anti-inflammatory effect can suppress tumor growth through immune regulation.

[0085] Therefore, the present invention provides a pharmaceutical composition for preventing or treating cancer comprising the peptide as an active ingredient, a use of the peptide for preventing or treating cancer, and a method for preventing or treating cancer comprising administering the peptide to a subject.

[0086] In one embodiment of the present invention, the cancer may be a solid cancer or a non-solid cancer, such as gastric cancer, breast cancer, lung cancer, liver cancer, blood cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, cutaneous or intraocular melanoma. melanoma), uterine sarcoma, ovarian cancer, rectal cancer, anal cancer, colon cancer, colorectal cancer, fallopian tube carcinoma, endometrial carcinoma, cervical cancer, small intestine cancer, endocrine cancer, thyroid cancer cancer), parathyroid cancer The cancer may be a cancer of the urethra, adrenal cancer, soft tissue tumor, urethral cancer, prostate cancer, bronchogenic cancer, bone marrow tumor, or lymphoma.

[0087] Preferably, the cancer may be, but is not limited to, breast cancer, colon cancer, colorectal cancer, lung cancer, ovarian cancer, pancreatic cancer, melanoma or lymphoma.

[0088] In the present invention, the peptide acts as a ligand that binds to the FPR2 receptor and can be used as a pharmaceutical composition for preventing or treating cancer. For more detailed information, see Kim et al.PLoS One.2012;7(1):e30522.doi:10.1371 / journal.pone.0030522.Epub 2012 Jan 25;Wang et al.Antioxidants(Basel).2021 Sep 14;10(9):1459.doi:10.3390 / antiox10091459.;Liu et al.Cancer Res.2013 Jan 15;73(2):550-60.doi:10.1158 / 0008-5472.CAN-12-2290.Epub 2012 Nov 8;Gilligan et al.Proc Natl Acad Sci USA2019 Mar 26;116(13):6292-6297.doi:10.1073 / pnas.1804000116.Epub 2019 Mar 12;Du et al.Ann Transl Med.2021 May;9(9):802.doi:10.21037 / atm-21-1873.Erratum in:Ann Transl Med.2022 Apr;10(7):428;Sulciner et al.J Exp Med.2018 Jan 2;215(1):115-140.doi:10.1084 / jem.20170681.Epub 2017 Nov 30;Zhang et al.Front Immunol.2017 Feb 2;8:71.doi:10.3389 / fimmu.2017.00071;Liotti et al.Cancers(Basel).2022 Jul 8;14(14):3333.doi:10.3390 / cancers14143333.

[0089] The term "prevention" as used herein means any action of suppressing or delaying immune diseases or cancer by administering the pharmaceutical composition according to the present invention, and the term "treatment" means any action of improving or making advantageous the symptoms of immune diseases or cancer by administering the pharmaceutical composition according to the present invention. In the present invention, the term "subject" means a subject in need of disease prevention or treatment, and more specifically, includes any mammal in need of immune disease or cancer prevention or treatment, including, without limitation, not only humans and primates, but also livestock such as cows, pigs, sheep, horses, dogs and cats, but is preferably a human.

[0090] The pharmaceutical composition containing the peptide as an active ingredient may be administered to a subject through any common route as long as the drug can reach the target tissue, and may be administered orally or parenterally. For example, it may be administered intraperitoneally, intravenously, intraarterially, intramuscularly, subcutaneously, intradermally, orally, topically (including skin application, infusion, and inhalation), transdermal, intrathecal and epidural, intranasal, intraocular, pulmonary, rectally, or vaginally, but is not limited thereto. In addition, the pharmaceutical composition containing the peptide as an active ingredient may be administered in any convenient pharmaceutical product form, for example, tablets, powders, granules, capsules, oral liquids, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. However, since the peptide is digested when administered orally, it is preferable that the oral composition is formulated so that the active agent is coated or protected from degradation in the stomach.

[0091] The pharmaceutical composition containing the peptide of the present invention may further contain pharma- ceutically and physiologically acceptable carriers, excipients, and diluents. In the present invention, the term "pharma-ceutically acceptable carriers, excipients, and diluents" refers to carriers, excipients, and diluents that do not stimulate the organism and do not inhibit the biological activity and properties of the administered compound. Examples of suitable carriers, excipients, and diluents that may be included in such compositions include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, glycerol, ethanol, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, amorphous cellulose, hypromellose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. When the composition is formulated into a pharmaceutical preparation, it may further contain a conventional dispersant, filler, extender, binder, disintegrant, surfactant, anti-agglomerating agent, lubricant, wetting agent, flavoring, emulsifier, preservative, freeze-dried preparation, etc.

[0092] In the present invention, the pharmaceutical composition may be administered in the form of an injection. The injection may be formulated as an intravenous injection or subcutaneous injection. In the case of a parenteral injection, it may contain components contained in a general injection composition. For example, the injection composition may contain a liquid carrier such as sterile water, water for injection, or physiological saline. In addition, it may further contain amino acids, sugars, lipids, vitamins, electrolytes, pH adjusters, stabilizers, osmotic pressure adjusters, or solubilizers.

[0093] In the present invention, when the pharmaceutical composition is administered locally, it may be formulated in the form of, for example, an ointment, gel, cream, lotion, etc. The form of local administration may be, but is not limited to, for example, application to the skin, instillation into the eyeball, transdermal permeation using a microneedle, intradermal injection, etc. For example, it may be preferable to apply the composition or attach a patch containing the composition to the skin. The composition may include, for example, a base material, an excipient, a lubricant, and a preservative. For example, when the pharmaceutical composition is administered in the form of an eye drop, it may further include a buffering agent, a viscosity agent, an isotonicity agent, a pH adjuster, and a solvent. In addition, when the pharmaceutical composition is administered in the form of an ointment for application to the skin, it may further include a gelling agent, a stabilizing agent, an emulsifying agent, and a suspending agent.

[0094] In addition, the pharmaceutical composition of the present invention may be applied differently depending on the purpose of administration and the disease. The amount of the active ingredient to be actually administered may be appropriately selected by those skilled in the art in consideration of various related factors, i.e., the disease to be treated, the severity of the disease, whether or not it is co-administered with other drugs, the activity of the drug, the sensitivity to the drug, the age, sex, weight, food, administration time, administration route, and administration ratio of the composition. The composition may be administered once or 1-3 times a day, although the dosage and administration route may be adjusted depending on the type and severity of the disease.

[0095] The pharmaceutical composition containing the peptide of the present invention as an active ingredient may be administered at a dose of 0.001 mg / kg to 1 g / kg, more preferably at a dose of 0.1 mg / kg to 100 mg / kg, while the dose may be appropriately adjusted depending on the age, sex and condition of the patient.

[0096] The present invention also provides an anti-inflammatory cosmetic composition containing the peptide as an active ingredient, a use of the peptide for suppressing inflammation, and a method for suppressing inflammation comprising applying the peptide to a subject.

[0097] The anti-inflammatory cosmetic composition containing the peptide of the present invention may be prepared in the form of a general emulsion or solubilization formulation, for example, a solution, an ointment for external use, a toner such as a softening toner or a nutritious toner, an emulsion such as a facial lotion or a body lotion, a cream such as a nutritious cream, a moisturizing cream or an eye cream, an essence, a cosmetic ointment, a spray, a gel, a pack, a sunscreen, a makeup base, a liquid type, a solid type or a spray type foundation, a powder, a makeup remover such as a cleansing cream, a cleansing lotion or a cleansing oil, a cleansing agent such as a cleansing foam, a soap or a body wash, an oil, a patch, etc., but is not limited thereto.

[0098] In the present invention, the cosmetic composition may further include one or more auxiliary agents and carriers commonly used in the field of cosmetology, such as pearling agents, thickeners, viscosity adjusters, pH adjusters, fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizing agents, foaming agents, fragrances, surfactants, water, ionic or non-ionic emulsifiers, fillers, sequestering and chelating agents, preservatives, preservatives, vitamins, blocking agents, moisturizing agents, essential oils, dyes, pigments, hydrophilic active agents, lipophilic active agents, lipid vesicles, or any other ingredients commonly used in cosmetics.

[0099] The advantages and features of the present invention, as well as the methods for achieving them, will become apparent from the following detailed description of the embodiments. However, the present invention is not limited to the embodiments disclosed below, and may be embodied in various different forms. However, the embodiments are provided to fully disclose the present invention and to fully inform those skilled in the art of the present invention of the scope of the invention, and the present invention is defined only by the scope of the claims.

[0100] Example 1: Production of immunomodulatory peptides

[0101] In the present invention, in order to synthesize novel peptides having superior immunomodulatory functions compared to the above peptides, they were synthesized using conventional amino acid synthesis methods (Umbarger, HE, Ann Rev Biochem., 47:533-606, 1978) and used as immunomodulatory candidate peptides. In addition, the present inventors chemically modified the N-terminus and / or C-terminus of the above-synthesized peptides. The sequences of the synthesized peptides are shown in Table 1.

[0102] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6]

[0103] <Experimental Example 1: Measurement of changes in calcium activity caused by peptides>

[0104] The present inventors observed changes in calcium ion permeability to confirm whether the immunomodulatory peptide candidates prepared according to one embodiment of the present invention activate the immune function of the living body. Specifically, the intracellular calcium ion concentration was measured to confirm whether the peptides activate FPR2.

[0105] 1) Cell and peptide preparation

[0106] The inventors used RBL cells that do not express FPR2 (RBL-WT) and RBL cells that overexpress FPR2 (RBL-FPR2), and utilized Fura-2 / AM, a dye with strong calcium binding properties, as a method for sensitively measuring the release of intracellular calcium ions.

[0107] Peptides dissolved in water or 50% DMSO at a concentration of 0.5 mM were prepared by diluting to appropriate concentrations in HBSS (Sigma, #55037C) + 20 mM HEPES buffer.

[0108] 2)Culture conditions

[0109] In this experiment, cells were cultured in DMEM (Lonza, #BE12-604F) medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin under conditions of 37°C and 5% CO2.

[0110] 3) Experimental Method

[0111] One day before measuring calcium activity, 3x105 cells were prepared per well in a 96-well flat clear bottom black plate (Costar, #3603). The same volume of Fura-2 QBT calcium kit (component A; Molecular Devices, #8197) was treated with the prepared cells per well. After incubation at 37℃ for 1 hour, the plate was incubated in a dark room at room temperature for 30 minutes. After Fura-2 treatment, the cells, the prepared peptides, and the dedicated tips were placed in each area of ​​the FlexStation 3 Multi-Mode Microplate Reader (Molecular Devices, #Flex3), and the changes in calcium activity due to the peptides were measured using SoftMax Pro7 Software (Molecular Devices, #FLEX3-INSTALL-OS).

[0112] 4) Experimental results

[0113] As a result, most of the peptides showed FPR2 activation potency, suggesting that the peptides according to the present invention can regulate the immune function of an individual through binding and activation of FPR2. The results of analyzing the FPR2 activation potency of the peptides were ranked according to numbers and summarized in Table 2 below. The criteria for each rank are shown in Table 3.

[0114] [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6]

[0115] [Table 3]

[0116] As shown in Table 2, the immunomodulatory peptide according to the present invention has an EC 50 EC50 values ​​for FPR1 were generally below 100 nM. 50 The value was confirmed to be 10,000 nM or more, confirming that the immunomodulatory peptide specifically binds to FPR2.

[0117] <Experimental Example 2: Measurement of cytokine suppression ability by peptides>

[0118] The present inventors have observed the cytokine suppression ability of the immunoregulatory peptide candidates prepared according to one embodiment of the present invention. Specifically, to confirm whether the peptides reduce intracellular inflammatory responses, the inventors measured the changes in intracellular cytokine gene expression under inflammatory conditions.

[0119] 1) Cell and peptide preparation

[0120] The present inventors used mouse macrophage RAW264.7 cell line and lipopolysaccharide (LPS; Sigma-Aldrich, #L2630), the main component of gram-negative bacterial cell walls, as a substance that induces an inflammatory response. Peptides were prepared in water at a concentration of 100 μM.

[0121] 2)Culture conditions

[0122] In this experiment, cells were cultured in DMEM (Lonza, #BE12-604F) medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin under conditions of 37°C and 5% CO2.

[0123] 3) Experimental Method

[0124] 2.5x10 per well in a 24-well plate (Costar, #3603) 5 After 24 hours, the prepared cells were treated with peptides at a concentration of 100 nM. After 30 minutes of incubation at 37°C, LPS at a concentration of 200 ng / ml was treated to induce inflammation. After 6 hours of incubation at 37°C, the cells were treated with TRIzol in the absence of the medium. TMCells were lysed using ELISA kit (Invitrogen, #15596018). RNA was obtained using the experimental method given, and cDNA was generated using reverse transcriptase (Promega, #M1705). Cytokine expression levels were then measured using SYBR Green (Enzynomics, RT530) and mouse primers for each inflammatory cytokine gene (TNFα, IL-6, IL-1β).

[0125] 4) Experimental results

[0126] As a result, it was confirmed that inflammatory cytokines (TNFα, IL-6, IL-1β) were reduced in most of the peptides compared to the control group. This suggests that the peptides according to the present invention can reduce inflammatory immune responses in individuals. The results of analyzing the changes in cytokine gene expression in the peptides were numerically graded and summarized in Table 4 below. The criteria for each grade are shown in Table 5.

[0127] [Table 4]

[0128] [Table 5]

[0129] In the following experimental examples, peptides having excellent effects were selected from the candidate immunomodulatory peptides, and the efficacy of the peptides was confirmed in animal models.

[0130] <Experimental Example 3: Confirmation of the efficacy of peptides in suppressing inflammatory responses at the individual level>

[0131] The present inventors observed the inflammatory response suppression efficacy at an individual level using the immunomodulatory peptide candidates prepared according to the embodiments of the present invention. Specifically, to confirm whether the peptides control inflammatory responses at an individual level, changes in cytokines were observed in a lipopolysaccharide (LPS; Sigma-Aldrich, #L2630)-induced acute peritonitis model.

[0132] 1) Induction of acute peritonitis model and peptide administration method

[0133] The present inventors used C57BL / 6 mice approximately 6 weeks old, and used LPS mixed with 5% mannitol (D-Mannitol; Sigma-Aldrich, #M8429) as an inducer of peritoneal inflammatory disease. The inflammatory substance was injected into the peritoneal cavity of the mice to induce peritoneal inflammatory disease, and the peptide was injected once into the peritoneal cavity immediately after the injection of the inflammatory substance.

[0134] 2) Disease evaluation method

[0135] After the end of peptide administration, blood and peritoneal lavage fluid were collected 6 hours or 24 hours later, and inflammatory factors such as cytokines were analyzed using the ELISA (Enzyme-Linked Immunosorbent Assay) technique.

[0136] 3) Experimental results

[0137] As a result, it was confirmed that inflammatory cytokines (TNFα, IL-6, IL-1β, IL-17a) were reduced in most individuals when the peptide was administered compared to the control group. This suggests that the peptide of the present invention can suppress inflammatory immune responses at the individual level. The results of analyzing the changes in cytokines when the peptide was administered are shown in Figure 1.

[0138] <Experimental Example 4: Confirmation of efficacy of peptides in improving rheumatoid arthritis>

[0139] The present inventors have observed the efficacy of immunomodulatory peptide candidates prepared according to one embodiment of the present invention in improving rheumatoid arthritis. Specifically, to confirm whether the peptides reduce intracellular inflammatory responses, changes in clinical scores and arthritis symptoms such as plantar edema in a collagen-induced arthritis (CIA) model were measured.

[0140] 1) Induction of collagen-induced arthritis model and peptide administration method

[0141] The inventors used DBA1 / J mice about 8-12 weeks old, and used a mixture of CFA (complete fraud adjuvant), which is a substance that induces an inflammatory reaction, and bovine Type II collagen as an arthritis inducer. The inflammatory substance was injected into the tail of the mice as a first injection (1 st injection), and about 2-3 weeks later, a second injection of the inflammatory agent ( nd Over the next few days, the peptide was intraperitoneally injected once every two days for a total of 10 times.

[0142] 2) Method for evaluating improvement of rheumatoid arthritis

[0143] Body weight and macroscopic changes were observed throughout the experimental period to confirm the overall health of the animals. Severity scores for arthritis and changes in thickness due to swelling in the soles and ankles were monitored throughout the experimental period. After the final substance administration, blood was collected to analyze inflammatory factors in the blood, such as cytokines, and tissues from the ankles were excised and paraffin blocks were prepared for tissue analysis using H&E (Hematoxylin and eosin) staining and immunohistochemistry.

[0144] 3) Experimental results

[0145] As a result, it was confirmed that the severity score of collagen-induced arthritis (CIA) was improved when the peptide was administered. This suggests that the peptide according to the present invention can improve arthritis, an inflammatory disease at the individual level. The results of analyzing the change in the severity score of the peptide are shown in FIG. 2, and the criteria for assigning the severity score are shown in Table 6.

[0146] [Table 6]

[0147] <Experimental Example 5: Confirmation of efficacy of peptide in improving dry eye syndrome>

[0148] The present inventors have observed the efficacy of immunomodulatory peptide candidates prepared according to one embodiment of the present invention in improving xerophthalmia. Specifically, to confirm whether the peptides improve the symptoms of xerophthalmia, changes in symptoms of xerophthalmia, such as the degree of corneal damage and corneal opacity, were measured in a benzalkonium chloride (BAC)-induced dry eye disease (DED) model.

[0149] 1) Induction of BAC-induced xerophthalmia model and peptide administration method

[0150] We treated C57BL / 6 mice aged about 12-16 weeks with BAC as a xerophthalmia inducer. BAC was administered topically by instillation into the right eye twice a day for a total of 16 days. For 4 days (total of 8 times), only BAC was administered, and from the 5th day, the peptide was also administered topically by instillation into the eye in the same manner 30 minutes after BAC administration.

[0151] 2) Evaluation method for xerophthalmia

[0152] During the experiment, changes were observed with the naked eye every day, and corneal damage was confirmed under a microscope using 0.2% fluorescein, a fluorescent indicator, on days 4, 8, 11, and 16 after the start of administration. The area stained with fluorescein indicates corneal damage, and the extent of corneal damage was evaluated by calculating the area of ​​staining. At the same time, the eyes were observed under a microscope and the degree of corneal opacity was scored.

[0153] 3) Experimental results

[0154] As a result, it was confirmed that the corneal damage and corneal opacity caused by BAC were improved when the peptide was administered. This suggests that the peptide according to the present invention can improve the symptoms of xerophthalmia in individuals. The results of analyzing the degree of corneal damage and corneal opacity caused by the peptide are shown in Figure 3.

[0155] <Experimental Example 6: Confirmation of the efficacy of peptides in improving colitis>

[0156] The present inventors have observed the efficacy of the immunomodulatory peptide candidates prepared according to one embodiment of the present invention in improving colitis. Specifically, to confirm whether the peptides reduce inflammatory responses occurring in the colon, changes in symptoms of colitis such as changes in individual weight and intestinal length were measured in a Dextran Sulfate Sodium (DSS)-induced colitis (IBD, Inflammatory bowel diseases) model.

[0157] 1) Induction of DSS-induced colitis model and peptide administration method

[0158] The present inventors used C57BL / 6 mice about 7-8 weeks old, dissolved 2.5% DSS in water as a colitis inducer, and allowed the mice to drink the water ad libitum. After 3 days, the existing water was discarded, and the mice were allowed to drink fresh 2.5% DSS water for another 2 days. After a total of 5 days, the water was replaced with fresh water. Starting the day after the 2.5% DSS was provided, 100 μl of peptide was intraperitoneally administered once a day.

[0159] 2) Method for evaluating colitis

[0160] Body weights were measured daily for the duration of the experiment, and after 7 days mice were euthanized with CO2 and intestinal lengths were measured.

[0161] 3) Experimental results

[0162] As a result, it was confirmed that administration of the peptide improved weight loss and restored intestinal length, which is shortened by inflammation. This suggests that the peptide of the present invention can improve the symptoms of colitis in individuals. The weight and intestinal length results of mice with colitis treated with the peptide are shown in Figure 4.

[0163] <Experimental Example 7: Effect of peptides on neuroinflammation>

[0164] The present inventors have observed the neuroinflammation suppression ability of the immunomodulatory peptide candidates prepared according to one embodiment of the present invention. Specifically, to confirm whether the peptides reduce inflammatory responses in neurons, the changes in the expression of cytokine genes in neurons under inflammatory conditions were measured.

[0165] 1) Cell and peptide preparation

[0166] The inventors used human neuronal cell line SK-N-MC and rat neuronal cell line DRG (dorsal root ganglion) neurons. In this experiment, the cells were cultured in Neuronal Cell Culture medium at 37°C and 5% CO2. Lipopolysaccharide (LPS; Sigma-Aldrich, #L2630), the main component of gram-negative bacterial cell walls, was used as a substance that induces an inflammatory response. Peptides were prepared by dissolving them in water at a concentration of 1 mM.

[0167] 2) Experimental Method

[0168] 2.5x10 per well of a 24-well plate (Costar, #3603) 5After 24 hours, the prepared cells were treated with peptides at a concentration of 1 μM. After 30 minutes of incubation at 37°C, LPS was treated at a concentration of 500 ng / ml to induce inflammation. After 24 hours of incubation at 37°C, the cells were treated with TRIzol in the absence of the medium. TM Cells were lysed using ELISA kit (Invitrogen, #15596018). RNA was obtained using the experimental method given, and cDNA was generated using reverse transcriptase (Promega, #M1705). Cytokine expression levels were then measured using SYBR Green (Enzynomics, RT530) and mouse primers for each inflammatory cytokine gene (IL-1β, IL-6).

[0169] 3) Experimental results

[0170] As a result, it was confirmed that most of the peptides reduced inflammatory cytokines (IL-1β, IL-6) compared to the control group. This suggests that the peptides according to the present invention can reduce neuroinflammatory responses in individuals. The results of analyzing changes in cytokine gene expression following treatment with the peptides are shown in FIG. 5.

[0171] <Experimental Example 8: Efficacy of peptides in improving degenerative brain diseases caused by neuroinflammation>

[0172] The present inventors have observed the efficacy of immunomodulatory peptide candidates prepared according to one embodiment of the present invention in improving brain diseases associated with neuroinflammation. Specifically, to confirm whether the peptide reduces Parkinson's disease based on its inhibitory effect on inflammatory responses in neurons, the inventors analyzed the changes in the number of dopamine neurons and the density of terminal fibers in brain tissue in an MPTP-induced Parkinson's disease animal model.

[0173] 1) Induction of MPTP-induced Parkinson's disease model and peptide administration method

[0174] The present inventors used 7-8 week old C57BL / 6 mice and used MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine hydrochloride), which induces neuroinflammatory reactions, as a brain lesion inducer. After inducing the disease by intraperitoneally administering 15 mg / kg MPTP four times, the peptide was intraperitoneally administered once a day for seven days, a total of seven times.

[0175] 2) Method for evaluating improvement of Parkinson's disease

[0176] The day after the final administration, the mice were anesthetized, perfused with phosphate buffered saline and fixative (4% paraformaldehyde) to remove blood and fix the tissue, and then the brains were removed. The removed brains were cryoblocked, sectioned, and stained for dopamine neurons using TH staining. The stained tissues were then photographed under a microscope, and the recovery of dopamine neuron numbers and nerve terminal fiber density was analyzed.

[0177] 3) Experimental results

[0178] As a result, it was confirmed that the density of nerve terminal fibers of dopamine neurons decreased by MPTP was restored when the immunomodulatory peptide was administered. This suggests that the peptide according to the present invention can improve Parkinson's disease by restoring dopamine neurons caused by neuroinflammation at the individual level through immune regulation through inflammation resolution. The results of analyzing the change in severity score of the peptide are shown in Figure 6.

[0179] <Experimental Example 9: Confirmation of tumor growth suppression ability of peptide>

[0180] The present inventors have observed the tumor growth suppression ability of the immunomodulatory peptide candidates prepared according to one embodiment of the present invention. Specifically, to confirm whether the peptides suppress tumor growth, the change in tumor size was measured in a tumor cell implantation animal model.

[0181] 1) Tumor cell transplantation model and peptide administration method

[0182] The present inventors used C57BL / 6 mice about 8 weeks old and CT-26 cell line as colon cancer cells. 5 The cells were implanted subcutaneously into the flank of mice and tumor growth was induced for approximately two weeks. Animals with uniformly sized tumors were selected and randomly assigned to drug treatment groups. After that, the peptide was intraperitoneally injected five times a week for two weeks, and the positive control drug 5-FU (5-Fluorouracil) was injected twice a week.

[0183] 2) Method for evaluating tumor growth inhibition

[0184] During the experimental period, body weight and macroscopic changes were observed to confirm the general health of the animals. The tumor's horizontal and vertical lengths were measured every 3-4 days, and the tumor was assumed to be an ellipsoid and its volume was calculated based on that ellipsoid. Changes in tumor volume were tracked throughout the experimental period to evaluate the efficacy of tumor growth inhibition. After administration of the final substance, blood was collected to analyze inflammatory factors or tumor-related factors, and paraffin blocks were prepared from the excised tumor tissues and subjected to tissue analysis using H&E (Hematoxylin and eosin) staining and immunohistochemistry techniques.

[0185] 3) Experimental results

[0186] As a result, it was confirmed that the administration of the peptide inhibited the growth of tumors formed by transplantation of the CT-26 cell line. This suggests that the peptide of the present invention can inhibit tumor growth through immune regulation based on its inflammation-reducing function. The results of analyzing the change in severity score of the peptide are shown in Figure 7.

[0187] The present invention has been described with reference to the above embodiments and experimental examples, but these are merely illustrative, and those skilled in the art will understand that various modifications and equivalent other embodiments and experimental examples are possible. Therefore, the true technical scope of the present invention should be determined by the technical spirit of the appended claims.

Claims

1. A peptide consisting of an amino acid sequence represented by X1-X2-X3-X4-X5-X6-m, said X1 is absent, arginine (R), lysine (K), histidine (H) or proline (P); said X2 is methionine (M) or tryptophan (W); The X3 is a basic amino acid, arginine (R) or lysine (K); said X4 is tyrosine (Y); X5 is a basic amino acid, such as arginine (R) or lysine (K), or an aromatic amino acid, such as tryptophan (W) or tyrosine (Y); said X6 is proline (P), tyrosine (Y) or valine (V); A peptide, wherein m is D-methionine.

2. said X1 is absent, arginine (R), lysine (K), histidine (H) or proline (P); said X2 is methionine (M) or tryptophan (W); The X3 is a basic amino acid, arginine (R) or lysine (K); said X4 is tyrosine (Y); said X5 is arginine (R) or lysine (K); said X6 is valine (V), tyrosine (Y) or proline (P); 2. The peptide according to claim 1, wherein m is D-methionine.

3. The X1 is absent or is a basic amino acid, arginine (R) or lysine (K); said X2 is methionine (M) or tryptophan (W); The X3 is a basic amino acid, arginine (R) or lysine (K); said X4 is tyrosine (Y); The X5 is tryptophan (W) as an aromatic amino acid; said X6 is valine (V), tyrosine (Y) or proline (P); 2. The peptide according to claim 1, wherein m is D-methionine.

4. The X1 is absent or is a basic amino acid, arginine (R) or lysine (K); said X2 is methionine (M) or tryptophan (W); The X3 is arginine (R) as a basic amino acid; said X4 is tyrosine (Y); said X5 is tyrosine (Y); said X6 is valine (V), tyrosine (Y) or proline (P); 2. The peptide according to claim 1, wherein m is D-methionine.

5. The peptide according to claim 1, wherein any one selected from the group consisting of an acetyl group, a butanoyl group, a hexanoyl group, and an octanoyl group is added to the N-terminus of the peptide.

6. The peptide according to claim 1, characterized in that the C-terminus of the peptide is amidated.

7. The peptide according to claim 1, characterized in that it is composed of any one of the amino acid sequences of SEQ ID NOs: 1 to 127.

8. A peptide consisting of an amino acid sequence represented by X1-X2-X3-X4-X5-X6-m, The X1 is a basic amino acid, arginine (R) or lysine (K); The X2 is tryptophan (W) as an aromatic amino acid; The X3 is a basic amino acid, arginine (R) or lysine (K); said X4 is tyrosine (Y); said X5 is arginine (R), lysine (K), leucine (L), asparagine (N), phenylalanine (F) or threonine (T); said X6 is isoleucine (I), threonine (T), tyrosine (Y) or valine (V); A peptide, wherein m is D-methionine.

9. A pharmaceutical composition for preventing or treating immune diseases or cancer, comprising the peptide according to any one of claims 1 to 8 as an active ingredient.

10. The immune diseases include atopic dermatitis, psoriasis, infantile eczema, sepsis, conjunctivitis, keratitis, dry eye syndrome, asthma, lung injury, lung inflammation, rheumatoid arthritis, ankylosing spondylitis, ulcerative colitis, Crohn's disease, and the like.

10. The pharmaceutical composition according to claim 9, wherein the disease is selected from the group consisting of inflammatory bowel disease, peritonitis, systemic sclerosis, neuroinflammation, Parkinson's disease and Alzheimer's disease.

11. An anti-inflammatory cosmetic composition comprising the peptide according to any one of claims 1 to 8 as an active ingredient.

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