KR20210147362A - A novel anti-inflammatory peptides derived nlrx1 and use thereof - Google Patents

Abstract

The present application provides an anti-inflammatory peptide comprising at least one selected from a group consisting of the amino acid sequence of SEQ ID NOs: 1 to 4 and modified amino acid sequences thereof, an anti-inflammatory composition comprising the same, and a pharmaceutical composition for preventing or treating inflammatory diseases. The anti-inflammatory peptide according to one embodiment of the present application has no cytotoxicity, and has an effect of inhibiting MAVS activation, immune/inflammatory activation reaction mechanism, inflammatory cytokines and inflammasome expression or activity, etc., so that the peptide can be applied to a composition for reducing inflammation or treating inflammatory diseases.

Description

Novel anti-inflammatory peptides derived from NLRX1 and uses thereof

The present application provides an anti-inflammatory peptide comprising at least one selected from the group consisting of the amino acid sequence of SEQ ID NOs: 1 to 4 and modified amino acid sequences thereof, an anti-inflammatory composition comprising the same, and a pharmaceutical composition for preventing or treating inflammatory diseases it's about

The proportion of the elderly continues to increase due to the development of medical technology and the extension of life expectancy following economic development. (Chang et al., 1994, Korean J. Gastroentrol., 26:907-918.; Heinzemann and Daser, 2002, Int. Arch. Allergy Immunol. 127:170-180.; Song et al. , 1998, Korean J. Intern. Med., 55:158-168.; Sung et al., 2012, J. Ethnopharmacol. 144:94-100.).

In general, the inflammatory response is an external stimulus such as bacterial or viral infection (pathogen-associated molecular pattern, PAMP) or an internal stimulus such as an in vivo metabolite following tissue damage (damage-associated molecular type, danger-associated molecular pattern). pattern, DAMP) as a defense mechanism of living tissues, such as TNF-α (tumor necrosis factor-α), IL-1β (interleukin-1β), IL-6 (interleukin-6) It is caused by the production of several cytokines and nitric oxide (NO). Lipopolysaccharide (LPS), also known as endotoxin, is an example of a representative pathogen-associated molecular type. It exists in the outer membrane of Gram-negative bacteria and is an intracellular transcription factor NF-κB (nuclear factor-κB) in macrophages or mononuclear cells. By inducing the activation of inflammatory cytokines, it induces gene expression of iNOS (inducible nitric oxide synthase) and COX-2 (cyclooxygenase-2) and generates inflammatory mediators. Therefore, substances that modulate the inflammatory response (e.g., the expression of iNOS, COX-2, or NF-κB, and the secretion of cytokines and nitric oxide, etc.) by the pathogen-associated molecular type or the damage-related molecular type are inflammatory It has recently attracted attention as a preventive and therapeutic agent for diseases (Dela Cruz and Kang, 2018, Mitochondrion, 41:37-44; Kim et al., 2013b, J. Korean Med. Ophthalmol. Otolaryngol. Dermatol., 26:54-64 .).

Materials currently used for anti-inflammatory purposes include nonsteroidal flufenamic acid, ibuprofen, benzydamine, indomethacin, and the like; Steroids include prednisolone, dexamethasone, betamethasone, and hydrocortisone, but these substances are highly toxic and cause serious side effects such as liver damage, cancer, and stroke. Restrictions follow. In addition, there are cases in which a problem of inducing severe immunosuppression occurs because it cannot selectively act on substances that cause inflammation. Accordingly, anti-inflammatory drugs extracted from natural substances have a weak effective concentration, and are cultivated in agricultural fields, etc. Therefore, there is a problem that the production cost is high.

In order to improve the above problems, a new concept anti-inflammatory agent is being developed as an alternative to the existing chemical anti-inflammatory agent or an anti-inflammatory agent using a natural product, and in particular, a lot of research is being done on the synthesis of peptides having anti-inflammatory activity.

However, in general, the synthesized peptide has very good anti-inflammatory activity and does not show cytotoxicity in in vitro experiments, but in fact, the anti-inflammatory effect is often insignificant in in vivo experiments.

This is due to several reasons, but mainly because physiological and anatomical conditions in vivo are very different from experimental conditions in vitro. First, in the presence of salt, which is a physiological condition, the activity of a peptide having a low positive charge is significantly inhibited. Second, because the peptide has a small molecular weight and size, it is almost absorbed by the kidney and excreted out of the body. Third, it is easily cleaved by protein and peptide degrading enzymes (protease and peptidase) present in all tissues, cells, body fluids, and blood in the living body, and thus loses its activity.

Accordingly, the present inventors have made intensive efforts to develop a substance exhibiting excellent anti-inflammatory activity while solving the above problems, and as a result, a peptide capable of economical mass production using 5 to 15 general amino acid residues has been developed, and the peptide is a cell By confirming that it does not show toxicity and exhibits excellent anti-inflammatory activity, the present invention has been completed.

The present application provides an anti-inflammatory peptide comprising at least one selected from the group consisting of the amino acid sequence of SEQ ID NOs: 1 to 4 and modified amino acid sequences thereof, an anti-inflammatory composition comprising the same, and a pharmaceutical composition for preventing or treating inflammatory diseases would like to provide

However, the problems to be solved by the present application are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

A first aspect of the present application provides an anti-inflammatory peptide comprising at least one selected from the group consisting of amino acid sequences of SEQ ID NOs: 1 to 4 and modified amino acid sequences thereof.

A second aspect of the present application provides a polynucleotide encoding the anti-inflammatory peptide of the present application.

A third aspect of the present application provides an anti-inflammatory composition comprising the anti-inflammatory peptide of the present application as an active ingredient.

A fourth aspect of the present application provides a pharmaceutical composition for preventing or treating inflammatory diseases, comprising the anti-inflammatory peptide of the present application as an active ingredient.

The anti-inflammatory peptide according to one embodiment of the present application has no cytotoxicity, and has an effect of inhibiting Mitochondrial Antiviral Signaling Protein (MAVS) activation, immune/inflammatory activation reaction mechanism, inflammatory cytokines and inflammasome expression or activity. Having a bar, the peptide can be applied to a composition for treating anti-inflammatory or inflammatory diseases.

1 is a diagram showing the structure of the novel anti-inflammatory peptide developed herein.
Figure 2 is a view showing the experimental results confirming the expression inhibitory effect on the IFN-β induced by polyinosinic: polycytidylic acid (polyIC) of the anti-inflammatory peptide of the present application.
3 is a view showing the experimental results confirming the cytotoxicity of the anti-inflammatory peptide of the present application.
4 is a view showing the experimental results confirming the expression inhibitory effect of the anti-inflammatory peptide of the present application on IL-1β induced by LPS or bacterial outer membrane vesicle (OMV).
5 is a view showing the experimental results confirming the expression inhibitory effect on the LPS-induced IL-6 and TNF-α of the anti-inflammatory peptide of the present application.
6 is a view showing the experimental results confirming the effect of inhibiting the expression of the anti-inflammatory peptide of the present application on IL-6 and TNF-α induced by OMV.

Hereinafter, embodiments of the present application will be described in detail with reference to the accompanying drawings so that those of ordinary skill in the art can easily implement them. However, the present application may be implemented in several different forms and is not limited to the embodiments described herein. And in order to clearly explain the present application in the drawings, parts irrelevant to the description are omitted, and similar reference numerals are attached to similar parts throughout the specification.

Throughout this specification, when a part is “connected” with another part, it is not only “directly connected” but also “indirectly connected” with a material such as another linker in between. cases are included.

Throughout this specification, when a part "includes" a component, it means that other components may be further included, rather than excluding other components, unless otherwise stated. As used throughout this specification, the terms “about”, “substantially”, etc. are used in a sense at or close to the numerical value when the manufacturing and material tolerances inherent in the stated meaning are presented, and are intended to enhance the understanding of the present application. To help, precise or absolute figures are used to prevent unfair use by unconscionable infringers of the stated disclosure. As used throughout this specification, the term “step of (to)” or “step of” does not mean “step for”.

Throughout this specification, the term “combination(s)” included in the expression of the Markush form means one or more mixtures or combinations selected from the group consisting of the components described in the expression of the Markush form, It means to include one or more selected from the group consisting of the above components.

Throughout this specification, reference to “A and/or B” means “A or B, or A and B”.

Hereinafter, embodiments and examples of the present application will be described in detail with reference to the accompanying drawings. However, the present application may not be limited to these embodiments and examples and drawings.

A first aspect of the present application provides an anti-inflammatory peptide comprising at least one selected from the group consisting of amino acid sequences of SEQ ID NOs: 1 to 4 and modified amino acid sequences thereof.

As used throughout this specification, the term “peptide” refers to a linear molecule formed by bonding amino acid residues to each other by a peptide bond (-CO-NH-). The peptides herein are prepared by chemical synthesis methods known in the art, particularly solid-phase synthesis techniques; Merrifield, J. Amer. Chem. Soc. 85:2149-54 (1963); Stewart, et al., Solid Phase Peptide Synthesis, 2nd. ed., Pierce Chem. Co.: Rockford, 111 (1984)) or liquid synthesis technology (US Patent No. 5,516,891) may be prepared, and the amino acid residues constituting the peptide of the present application are It may be a natural or non-natural amino acid residue.

In one embodiment of the present application, the peptide may be derived from a nucleotide-binding oligomerization domain, leucine-rich repeat containing X1 (NLRX1) protein, and specifically, the peptide is human-derived represented by the amino acid sequence of SEQ ID NO: 5 In the NLRX1 protein, peptides consisting of amino acids 77 to 86 (MQPX-76), peptides consisting of amino acids 511 to 520 (MQPX-519), peptides consisting of amino acids 751 to 760 (MQPX-751), and positions 779 to 789 It may be one or more selected from the group consisting of a peptide consisting of amino acids (MQPX-779).

In one embodiment of the present application, the peptide is selected from the group consisting of the amino acid sequence of SEQ ID NO: 1, the amino acid sequence of SEQ ID NO: 2, the amino acid sequence of SEQ ID NO: 3, the amino acid sequence of SEQ ID NO: 4, and modified amino acid sequences thereof It may include one or more, and specifically may include the amino acid sequence of SEQ ID NO: 2.

In one embodiment of the present application, the modified amino acid sequence may maintain or exhibit improved activity without changing the main activity, a part of the amino acid sequence is mutated by natural or artificial mutation, or the amino acid One or more of the amino acids constituting the sequence may be substituted with other types of amino acids. Specifically, the modified amino acid sequence is, at least one of the amino acids constituting the amino acid sequence of SEQ ID NOs: 1 to 4 gamma amino butanoic acid, glutamine (Gln, Q), glutamic acid (Glu, E), glycine (Gly, G ), lysine (Lys, K), leucine (Leu, L), methionine (Met, M), (Val, V), serine (Ser, S), selenomethionine, selenocysteine (Sec, U), cysteine (Cys, C), citrulline, arginine (Arg, R), asparagine (Asn, N), aspartic acid (Asp, D), alanine (Ala, A), ornithine, isoleucine (Ile, I) , Taurine, Threonine (Thr, T), Tryptophan (Trp, W), Tyrosine (Tyr, Y), Phenylalanine (Phe, F), Proline (Pro, P), Pyrrolysine (Pyr, O), Histidine (His) , H) and may be substituted with one or more selected from the group consisting of non-natural amino acids.

Peptides according to one embodiment of the present application are 80% or more, 85% or more, 90% or more, 91% or more, It may include a peptide exhibiting 92% or more, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more homology. If an amino acid sequence having the same or corresponding biological activity as the peptide of SEQ ID NO: substantially described as a sequence having homology with the above sequence, some sequences having an amino acid sequence deleted, modified, substituted or added are also of the present application. It is self-evident that it is included in the category.

As used throughout this specification, the term "homology" refers to the degree of correspondence with a given amino acid sequence or polynucleotide sequence and may be expressed as a percentage. In the present specification, a homologous sequence having the same or similar activity to a given amino acid sequence or polynucleotide sequence is expressed as "% homology". For example, standard software that calculates parameters such as score, identity and similarity, specifically BLAST 2.0, or hybridization used under defined stringent conditions Appropriate hybridization conditions defined, which can be confirmed by comparing sequences by experiment, are within the scope of the art, and methods well known to those skilled in the art [eg, J. Sambrook et al., Molecular Cloning, A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory press, Cold Spring Harbor, New York, 1989; F.M. Ausubel et al., Current Protocols in Molecular Biology, John Wiley & Sons, Inc., New York]. As used throughout this specification, the term "stringent conditions" refers to conditions that allow specific hybridization between polynucleotides. For example, such conditions are specifically described in, eg, J. Sambrook et al., supra.

As used throughout this specification, the term “anti-inflammatory” refers to an action that inhibits or reduces inflammation, and the term “inflammation” is a protective reaction that occurs in the body when a living tissue is damaged, causing an inflammatory disease is the cause Therefore, the anti-inflammatory peptide of the present application can be used for the prevention, treatment or improvement (relief of symptoms) of an inflammatory disease by exhibiting an activity of inhibiting or reducing inflammation.

In one embodiment of the present application, the peptide may further include a cell-penetrating peptide, but is not limited thereto.

As used throughout this specification, the term "cell-permeable peptide" is a peptide having the ability or property to penetrate a cell membrane and penetrate into a cell, and may indicate cell-permeability and/or skin permeability.

In one embodiment of the present application, the cell-penetrating peptide may further include a part or all of a sequence derived from a conventionally known cell-penetrating peptide or a skin-penetrating peptide, for example, dNP2, penetratin, Tat, transpotan, MAP, KALA, P1, MPG, Pep-1, Arg (7, 8, 9, 10, 11), hCT, pVEC, SPEH, YARA, WLR, VP22, MTS, FHV coat, and combinations thereof It may be selected from the group consisting of, and specifically, may be an Arg(8) peptide (R8 peptide), but is not limited thereto.

In one embodiment of the present application, the cell-penetrating peptide may be included in the anti-inflammatory peptide more than once, specifically, the cell-penetrating peptide is included twice, three times, five times, seven times or ten times. may be, but is not limited thereto.

In one embodiment of the present application, the cell-penetrating peptide may be linked to the N-terminus or the C-terminus of the anti-inflammatory peptide, and specifically may be linked to the N-terminus, but inhibits the pharmacological activity of the peptide However, it is not limited thereto as long as it can improve cell permeability or skin permeability.

In one embodiment of the present application, the cell-penetrating peptide may be linked to the anti-inflammatory peptide through a linker, or may be directly linked. In addition, the linker may be cleaved or degraded by various biological and chemical actions such as enzyme action in cells or in the skin, and the cell-penetrating peptide and the anti-inflammatory peptide according to the cleavage or degradation of the linker are the purpose They may be separated from each other in cells or in the skin.

In one embodiment of the present application, the amino acids constituting the anti-inflammatory peptide may be L-type, D-type or DL-type, and specifically may be D-type, but as long as it does not affect the activity of the peptide, it is not limited thereto. .

In one embodiment of the present application, the N-terminus or C-terminus of the anti-inflammatory peptide is an acetyl group, a fluorenyl methoxy carbonyl group, a formyl group, a palmitoyl group, a myristyl group, a stearyl group and polyethylene glycol ( A protecting group selected from the group consisting of polyethylene glycol, PEG) may be bound, and the protecting group is not limited thereto as long as it does not affect the activity of the peptide.

In one embodiment of the present application, the peptide or amino acids constituting the same may be acetylated or amination as well as the above protecting group bond, and the modification greatly improves the stability of the peptide, The stability means not only in vivo stability, but also storage stability (eg, room temperature storage stability), and the above-mentioned protecting group may act to protect the peptide of the present application from attack by protease in vivo.

In one embodiment of the present application, the peptide may inhibit MAVS protein aggregation and/or function in vivo.

As used throughout this specification, the term "MAVS (mitochondrial antiviral-signaling) protein" is a signaling protein essential for anti-viral innate immunity, and is present in the outer membrane of mitochondria, peroxisome, and endoplasmic reticulum (ER). is known to do The MAVS protein is activated by forming an aggregate (aggregate) by a cytoplasmic protein group that detects the presence of a virus upon viral infection, and the activated MAVS induces an immune response in a manner that induces secretion of interferon and cytokines. However, if the MAVS protein is continuously activated and the production of interferon and cytokines is excessively increased, it can rather attack cells in the body and cause various pathological abnormalities and immune diseases. Therefore, it is necessary to properly regulate the activation of the MAVS protein.

As used throughout this specification, the term "NLRX1 (nucleotide-binding oligomerization domain, leucine-rich repeat containing X1) protein" is known as an inhibitor that inhibits the intracellular function of MAVS protein, a signaling protein essential for antiviral innate immunity. , as it is the only pattern recognition receptor present in the outer mitochondrial membrane among several pattern recognition receptors (PRR) in cells, it is a key protein that connects the innate immune response and mitochondrial function. Therefore, it is possible to modulate the immune response through activation of the MAVS protein by increasing the intracellular expression of NLRX1 or using a substance that can replace the function of NLRX1.

In one embodiment of the present application, the anti-inflammatory peptide can inhibit the inflammatory/immune activation response mechanism and inflammatory/immune response induced by MAVS activation, through inhibition of MAVS aggregation or activation, as well as by MAVS aggregation It is possible to treat, prevent or ameliorate symptoms or diseases that may develop.

In one embodiment of the present application, the peptide may inhibit the expression, generation, activity, etc. of an inflammatory cytokine or inflammasome or inhibit the proliferation of inflammatory cells, specifically, the inflammatory cytokine is IFN It may be one or more selected from the group consisting of -β, IL-1β, IL-6 and TNF-α, but is not limited thereto as long as it is a substance known to cause an inflammatory response in the art.

As used throughout this specification, the term "inflammasome" refers to a substance that induces the maturation of inflammatory cytokines such as IL-1 related to innate immune defenses such as infection or stress of cells, It is a caspase-1-activating protein complex, 1) a sensor protein, NLRP3 (NOD-like receptor family, pyrin domain-containing 3), 2) an adapter protein, ASC (adaptor protein) It is composed of an apoptosis-associated spec-like protein containing a caspase-recruitment domain) and 3) an effector protein, inactive caspase-1. The components of the inflammasome are assembled when infection of microorganisms such as bacteria or viruses or tissue injury occurs. -1 (active caspase-1). It is known that the converted activated form of casphase-1 cleaves the precursor form of IL-1β or IL-18 to produce activated IL-1β or IL-18, and secretes it out of the cell to perform the host's innate immune defense function. have.

In one embodiment of the present application, it was confirmed that the anti-inflammatory peptide of the present application inhibits the expression level of various inflammatory cytokines and inflammasome, based on this, the anti-inflammatory peptide of the present application can effectively inhibit the inflammatory response. Able to know.

In one embodiment of the present application, the peptide may inhibit a mechanism of an inflammatory / immune activation response, and the mechanism is caspase-1, interferon regulatory factor 3 (IRF3) , or nuclear factor kappa-bi (nuclear factor kappa-light-chain-enhancer of activated B cells, NF-κB), but not limited thereto .

In one embodiment of the present application, the peptide may inhibit inflammation induced by bacteria or viruses, specifically, to inhibit inflammation induced by pathogen-associated molecular type (PAMP) derived from bacteria or viruses. can In addition, it may be to inhibit inflammation caused by damage-associated molecular type (DAMP) secreted out of cells through cell damage caused by bacterial or viral infection.

As used throughout this specification, the term "pathogen-associated molecular pattern (PAMP)" refers to a molecule that originates from a pathogen and causes an immune response, and the pattern recognition receptor possessed by the immune system responds to a specific molecular type and exhibits phagocytosis. Because it induces the formation of antibodies or eliminates the infectious agent through the The pathogen-associated molecular types are endotoxin, exotoxin, LPS (lipopolysaccharide), LTA (lipoteichoic acid), MDP (muramyl dipeptide), nigericin, dsRNA (polyIC, etc.), dsDNA (polydAdT) etc.), outer membrane vesicle (OMV), and flagellin, etc., but is not limited thereto, as long as it is a substance known to be derived from a pathogen and cause an inflammatory/immune response in the art.

As used throughout this specification, the term "danger-associated molecular pattern (DAMP)" is an intracellular metabolite or protein that is secreted out of the cell by cell damage and causes an immune response in surrounding cells. . The damage-related molecular type may be ATP, histone protein, HMGB1 (high mobility group box 1), mtDNA (mitochondrial DNA), uric acid, etc. Any known material is not limited thereto.

In one embodiment of the present application, the peptide can inhibit an inflammatory response or treat or alleviate symptoms of a disease caused by an inflammatory response, specifically, the peptide is an anti-inflammatory pharmaceutical composition, a food composition, a cosmetic composition , may be included in various compositions such as a health functional food composition, a feed composition.

A second aspect of the present application provides a polynucleotide encoding the anti-inflammatory peptide of the present application. The content overlapping with the first aspect also applies to the polynucleotide of the second aspect.

As used throughout this specification, the term “polynucleotide” refers to a high molecular material to which nucleotides are bound, and refers to DNA encoding genetic information.

In one embodiment of the present application, the polynucleotide may include a nucleotide sequence encoding at least one of the amino acid sequences of SEQ ID NO: 1 to SEQ ID NO: 5.

In one embodiment of the present application, the nucleotide sequence encoding the anti-inflammatory peptide is not only the nucleotide sequence encoding the amino acid described in each SEQ ID NO: 80% or more, specifically 90% or more, more specifically As a nucleotide sequence exhibiting 95% or more, more specifically 98% or more, and most specifically, 99% or more homology, as long as it is a nucleotide sequence encoding a protein that exhibits substantially the same or corresponding efficacy as each peptide, there is no limitation include In addition, if the amino acid sequence having the same or corresponding biological activity as the peptide of SEQ ID NO: substantially described as a sequence having homology with the above sequence, some sequences are deleted, modified, substituted or added to the amino acid sequence of the present invention as well. It is obvious that it is included in the category of In addition, the polynucleotide encoding the peptide is within the range that does not change the amino acid sequence of the peptide expressed from the coding region in consideration of the codon preferred in the organism in which the peptide is to be expressed due to codon degeneracy. Various modifications can be made to the coding region in . Accordingly, the polynucleotide may be included without limitation as long as it is a polynucleotide sequence encoding each peptide. In addition, if a probe that can be prepared from a known sequence, for example, a sequence encoding a protein having the activity of the peptide by hybridization under stringent conditions with a sequence complementary to all or part of the polynucleotide sequence, is not limited, may be included.

As used herein, the term “stringent conditions” refers to conditions that allow specific hybridization between polynucleotides. These conditions are specifically described in the literature (see, eg, Sambrook et al., supra, 9.50-9.51, 11.7-11.8). For example, genes with high homology between genes having homology of 40% or more, specifically 90% or more, more specifically 95% or more, still more specifically 97% or more, and particularly specifically 99% or more homology. Conditions that hybridize with each other and do not hybridize with genes with lower homology, or wash conditions of conventional southern hybridization at 60° C., 1X SSC, 0.1% SDS, specifically 60° C., 0.1X SSC , 0.1% SDS, more specifically at a salt concentration and temperature equivalent to 68°C, 0.1X SSC, 0.1% SDS, washing once, specifically 2 to 3 times. Hybridization requires that two polynucleotides have complementary sequences, although mismatch between bases is possible depending on the stringency of hybridization. The term “complementary” is used to describe the relationship between nucleotide bases capable of hybridizing to each other. For example, with respect to DNA, adenosine is complementary to thymine and cytosine is complementary to guanine. Accordingly, the present application may also encompass substantially similar polynucleotide sequences as well as isolated polynucleotide fragments complementary to the overall sequence.

Specifically, polynucleotides having homology can be detected using hybridization conditions including a hybridization step at a Tm value of 55° C. and using the conditions described above. In addition, the Tm value may be 60 °C, 63 °C, or 65 °C, but is not limited thereto and may be appropriately adjusted by those skilled in the art according to the purpose. The appropriate stringency for hybridizing polynucleotides depends on the length of the polynucleotides and the degree of complementarity, and the parameters are well known in the art (see Sambrook et al., supra, 9.50-9.51, 11.7-11.8).

As another aspect of the second aspect of the present application, there is provided an expression vector comprising the polynucleotide.

As used throughout this specification, the term "expression vector" is a recombinant vector that can be introduced into a suitable host cell to express a target protein, and refers to a genetic construct comprising essential regulatory elements operably linked to express a gene insert. .

As used throughout this specification, the term “operably linked” means that a nucleic acid expression control sequence and a nucleic acid sequence encoding a target protein are functionally linked to perform a general function. The operative linkage with the recombinant vector can be prepared using genetic recombination techniques well known in the art, and site-specific DNA cleavage and ligation can be facilitated using enzymes generally known in the art. have.

A suitable expression vector of the present disclosure may include a signal sequence for membrane targeting or secretion in addition to expression control elements such as a promoter, initiation codon, stop codon, polyadenylation signal and enhancer. The start codon and stop codon are generally considered to be part of the nucleotide sequence encoding the immunogenic target protein, and must be functional in the subject when the genetic construct is administered and must be in frame with the coding sequence. Common promoters can be constitutive or inducible, lac, tac, T3 and T7 promoters for prokaryotic cells, simian virus 40 (SV40) for eukaryotic cells, mouse mammary tumor virus (MMTV) promoter, human immunodeficiency virus (HIV), for example the long terminal repeat (LTR) promoter of HIV, Moloney virus, cytomegalovirus (CMV), Epstein Barr virus (EBV), Loes sarcoma virus (RSV) promoter, as well as β- promoters derived from actin promoter, human hemoglobin, human muscle creatine, and human metallothionein, but are not limited thereto.

In addition, the expression vector may include a selectable marker for selecting a host cell containing the vector. The selectable marker is for selecting cells transformed with the vector, and markers conferring a selectable phenotype such as drug resistance, auxotrophicity, resistance to cytotoxic agents or expression of surface proteins may be used. In an environment treated with a selective agent, only the cells expressing the selection marker survive, so the transformed cells can be selected. In addition, when the vector is a replicable expression vector, it may include a replication origin, which is a specific nucleic acid sequence from which replication is initiated.

As a recombinant expression vector for inserting a foreign gene, various types of vectors such as plasmids, viruses, and cosmids can be used. The type of recombinant vector is not particularly limited as long as it functions to express a desired gene and produce a desired protein in various host cells of prokaryotic and eukaryotic cells. A vector capable of producing a large amount of a foreign protein in a form similar to that of a can be used.

In order to express the anti-inflammatory peptides according to the present disclosure, various combinations of hosts and vectors may be used. Expression vectors suitable for eukaryotic hosts may include, but are not limited to, expression control sequences derived from SV40, bovine papillomavirus, adenovirus, adeno-associated virus, cytomegalovirus and retrovirus, and the like. Expression vectors that can be used in bacterial hosts are not limited thereto, but bacterial plasmids obtained from Escherichia coli , including pET, pRSET, pBluescript, pGEX2T, pUC, col E1, pCR1, pBR322, pMB9 or derivatives thereof, etc. , phage DNA exemplified by phage lambda derivatives such as plasmids with a wider host range such as RP4, λgt10, λgt11 or NM989, and other DNA phages such as M13 and filamentous single-stranded DNA phages etc. may be included. For yeast cells, a 2°C plasmid or a derivative thereof may be used, and for insect cells, pVL941 or the like may be used.

As another aspect of the second aspect of the present application, there is provided a transformant other than a human comprising the expression vector.

As used throughout this specification, the term "transformant" may be a cell of a host into which the expression vector can be introduced. Specifically, the transformant of the present application may be a transformant other than a human, but is not limited thereto.

Suitable host cells for introduction of the vector include E. coli, Bacillus subtilis , Streptomyces sp. , Pseudomonas sp. , Proteus mirabilis (Proteus mirabilis) or Staphylococcus It may be a prokaryotic cell such as Staphylococcus sp. In addition, fungi such as Aspergillus sp. , Pichia pastoris , Saccharomyces cerevisiae , Schizosaccharomyces sp. or Neuro It may be a cell of a yeast such as Neurospora crassa , other lower eukaryotic cells, or a higher eukaryote such as a cell of a plant or insect. In addition, the cells may be mammalian, specifically monkey kidney cells 7 (COS7) cells, NSO cells, SP2/0, Chinese hamster ovary (CHO) cells, W138, young hamster kidney ( BHK: baby hamster kidney) cells, MDCK, myeloma cell line, HuT 78 cells or HEK293 cells may be used, but the present invention is not limited thereto.

The transformation method of the present application includes any method for introducing a nucleic acid into an organism, cell, tissue or organ, and may be performed by selecting a suitable standard technique according to a host cell known in the art. Specifically, electroporation, protoplast fusion, calcium phosphate (CaPO ₄ ) precipitation, calcium chloride (CaCl ₂ ) precipitation, agitation using silicon carbide fiber, agrobacterium mediated transformation, PEG, dextran sulfate, lipoprotein Pectamine and drying/suppression mediated transformation methods, etc. are included, but are not limited thereto.

As another aspect of the second aspect of the present application, there is provided a method for producing an anti-inflammatory peptide comprising the step of culturing the transformant.

The method for producing the anti-inflammatory peptide includes culturing the transformant of the present application, specifically, inserting a polynucleotide sequence encoding the anti-inflammatory peptide into the vector to prepare an expression vector; preparing a transformant by introducing the expression vector into a host cell; culturing the transformant; and isolating and purifying the anti-inflammatory peptide from the cultured transformant.

More specifically, a large amount of peptides can be produced by culturing the transformant in a nutrient medium, and the medium and culture conditions tolerated depending on the host cell can be appropriately selected and used. In culture, conditions such as temperature, pH of the medium, and culture time can be appropriately adjusted to be suitable for cell growth and mass production of proteins.

As described above, the recombinantly produced peptide or protein may be recovered from the medium or cell lysate. If it is membrane-bound, it can be released from the membrane by using a suitable surfactant solution (eg Triton-X100) or by enzymatic cleavage. Cells used for expression of anti-Oscar antibody or fragment thereof may be disrupted by various material or chemical means, such as freeze-thaw acclimatization, sonication, mechanical disruption, or cytolytic agents, and isolate by conventional biochemical separation techniques; Purification is possible (Sambrook et al., Molecular Cloning: A laborarory Manual, 2nd Ed., Cold Spring Harbor Laboratory Press (1989); Deuscher, M., Guide to Protein Purification Methods Enzymology, Vol. 182. Academic Press. Inc.) ., San Diego, CA (1990)). Electrophoresis, centrifugation, gel filtration, precipitation, dialysis, chromatography (ion exchange chromatography, affinity chromatography, immunosorbent chromatography, size exclusion chromatography, etc.), isoelectric focusing and various variations and complex methods thereof are used. possible, but not limited thereto.

A third aspect of the present application provides an anti-inflammatory composition comprising the anti-inflammatory peptide of the present application as an active ingredient. The contents overlapping with the first aspect and the second aspect are equally applied to the composition of the third aspect.

In one embodiment of the present application, the anti-inflammatory composition may be used as a pharmaceutical, quasi-drug, cosmetic, food and feed composition.

In one embodiment of the present application, the pharmaceutical composition may be prepared as an oral dosage form or parenteral dosage form according to the route of administration by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. Here, "pharmaceutically acceptable" means that it does not inhibit the activity of the active ingredient and does not have toxicity beyond what the application (prescription) target can adapt.

In one embodiment of the present application, the pharmaceutical composition is in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, or sterile injection solutions, respectively, according to a conventional method. It may be formulated and used, but may not be limited thereto.

In one embodiment of the present application, when formulating the pharmaceutical composition, it may be prepared using a diluent or excipient such as a generally used filler, extender, binder, wetting agent, disintegrant, or surfactant, but is not limited thereto. it may not be

In one embodiment of the present application, when the pharmaceutical composition is prepared as an oral dosage form, powders, granules, tablets, pills, dragees, capsules, liquids, gels, It can be prepared in the form of syrup, suspension, wafer, and the like. Examples of suitable pharmaceutically acceptable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol, starches such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, Cellulose such as sodium carboxymethylcellulose and hydroxypropylmethylcellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable and oil. In the case of formulation, if necessary, the formulation may include a diluent and/or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, and a surfactant.

In one embodiment of the present application, when the pharmaceutical composition is prepared for parenteral use, it may be formulated in the form of injections, transdermal administrations, nasal inhalants and suppositories together with a suitable carrier according to methods known in the art. In the case of formulation for injection, suitable carriers include sterile water, ethanol, polyols such as glycerol or propylene glycol, or mixtures thereof, preferably Ringer's solution, PBS (phosphate buffered saline) containing triethanolamine, or sterile for injection. Water, an isotonic solution such as 5% dextrose, etc. may be used. When formulated for transdermal administration, it may be formulated in the form of an ointment, a cream, a lotion, a gel, an external solution, a pasta agent, a liniment agent, an air roll, and the like. In the case of nasal inhalants, it can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, and the like. witepsol), tween 61, polyethylene glycols, cacao fat, laurin fat, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, sorbitan fatty acid esters, etc. may be used.

In one embodiment of the present application, the pharmaceutical composition may be administered in a pharmaceutically effective amount, and the term "pharmaceutically effective amount" refers to treating or treating a disease at a reasonable benefit/risk ratio applicable to medical treatment or prevention. It means an amount sufficient to prevent, and the effective dose level is determined by the severity of the disease, the activity of the drug, the patient's age, weight, health, sex, the patient's sensitivity to the drug, the time of administration of the composition of the present invention used, the route of administration, and The rate of excretion may be determined according to factors well known in the medical arts, including the duration of treatment, the drug used in combination with or concurrently with the composition of the present application, and other factors well known in the medical arts. The pharmaceutical composition of the present application may be administered alone or in combination with a component known to exhibit a therapeutic effect on a known intestinal disease. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects.

In one embodiment of the present application, the dosage of the pharmaceutical composition can be determined by those skilled in the art in consideration of the purpose of use, the degree of addiction of the disease, the patient's age, weight, sex, history, or the type of substance used as an active ingredient, etc. have. For example, the pharmaceutical composition of the present invention may be administered in an amount of about 0.1 ng to about 1,000 mg/kg, preferably 1 ng to about 100 mg/kg per adult, and the frequency of administration of the composition of the present invention is particularly limited thereto. However, it can be administered once a day or administered several times by dividing the dose. The dosage or frequency of administration is not intended to limit the scope of the present application in any way.

A fourth aspect of the present application provides a pharmaceutical composition for preventing or treating inflammatory diseases, comprising the anti-inflammatory peptide of the present application as an active ingredient. The contents overlapping with the first to third aspects are equally applied to the composition of the fourth aspect.

As used throughout this specification, the term “treatment” refers to any action in which the symptoms of an inflammatory disease are improved or beneficially changed by administration of the composition of the present application.

As used throughout this specification, the term “prevention” refers to any action in which an inflammatory disease or the possibility of its onset is suppressed or delayed by administration of the composition of the present application.

As used throughout this specification, the term "inflammatory disease" refers to a local or systemic biodefense reaction against external physical or chemical stimuli or external infectious agents such as bacteria, bacteria, fungi, viruses, and various allergens, or autoimmunity. It can be defined as a pathological symptom caused by an inflammatory response. Such an inflammatory response involves the activation of various inflammatory mediators and immune cell-related enzymes (eg, iNOS, COX-2, etc.), secretion of inflammatory mediators (eg, secretion of NO, TNF-α, IL-6, etc.), infiltration of body fluids, It is accompanied by a series of complex physiological reactions such as cell migration and tissue destruction, and is externally manifested by symptoms such as erythema, pain, edema, fever, and deterioration or loss of specific body functions. Since the inflammatory disease can be acute, chronic, ulcerative, allergic or necrotic, whether it is acute, chronic, ulcerative, allergic or Regardless of whether it is necrotic or not.

In one embodiment of the present application, the inflammatory disease is sepsis, septic shock, systemic inflammatory response syndrome, acute respiratory distress syndrome, asthma, allergic and non-allergic rhinitis , chronic and acute rhinitis, chronic and acute gastritis or enteritis, ulcerative gastritis, acute and chronic nephritis, acute and chronic hepatitis, chronic obstructive pulmonary disease, idiopathic pulmonary fibrosis, irritable bowel syndrome, inflammatory pain, migraine , headache, back pain, fibromyalgia, myofascial disease, viral infection (such as hepatitis C infection), bacterial infection, fungal infection, burns, surgical or dental wounds, prostaglandin E excess syndrome, atherosclerosis Sclerosis, gout, arthritis, rheumatoid arthritis, ankylosing spondylitis, Hodgkin's disease, pancreatitis, conjunctivitis, iritis, scleritis, uveitis, dermatitis, atopic dermatitis, eczema, systemic lupus erythematosus (SLE) and multiple sclerosis It may be one or more selected from the group consisting of, if it is a disease caused by an inflammatory response, specifically, a disease caused by an inflammatory response caused by a bacterial or viral infection, it is not limited thereto.

A fifth aspect of the present application provides a method for preventing or treating inflammatory or inflammatory diseases, comprising administering to an individual the anti-inflammatory composition or the pharmaceutical composition for preventing or treating inflammatory diseases of the present application. The contents overlapping with the first to fourth aspects are equally applied to the method of the fifth aspect.

As used throughout this specification, the term "individual" may include, without limitation, mammals, farmed fish, and the like, including mice, livestock, humans, and the like, in which an inflammatory reaction or an inflammatory disease develops or is at risk of developing.

In one embodiment of the present application, the subject may be other than a human.

In one embodiment of the present application, in the method, a pharmaceutically effective amount of the composition may be administered to prevent or treat inflammation or inflammatory disease, the inflammatory response or the degree of progression of the inflammatory disease, the age, weight, and symptoms of the patient may vary depending on various factors, such as the nature and extent of treatment, the type of current treatment, the number of treatments, the dosage form, and route, and may be easily determined by experts in the field. The composition of the present application may be administered together or sequentially with the aforementioned pharmacological or physiological components, and may be administered in combination with an additional conventional therapeutic agent, or may be administered sequentially or simultaneously with the conventional therapeutic agent. Such administration may be single or multiple administrations. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, and can be easily determined by a person skilled in the art.

As used throughout this specification, the term "administration" means introducing a predetermined substance to an individual by an appropriate method, and the administration route of the composition of the present application may be administered through any general route as long as it can reach the target tissue. have. Intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, intrapulmonary administration, may be administered intrarectally, but is not limited thereto. However, when administered orally, since the protein is digestible, it is preferable that the oral composition be formulated to coat the active agent or to protect it from degradation in the stomach. In addition, the composition may be administered by any device capable of transporting the active agent to a target cell.

Hereinafter, the present invention will be described in more detail through examples of the present application, but the following examples are merely illustrative to aid understanding of the present application, and the content of the present application is not limited to the following examples.

[Example]

Example 1: Novel peptide synthesis with anti-inflammatory effect

In order to develop anti-inflammatory peptides that can alleviate inflammatory responses by inhibiting signal transduction through MAVS protein, which is known as an important hub protein for innate immune response mechanism, MAVS signal transduction in the following way A peptide sequence for inhibitory anti-inflammatory was derived.

Specifically, the NLRX1 protein binds to the MAVS protein that is activated by inflammatory stimulation by a pathogen-associated molecular type or an injury-associated molecular type, and inhibits the MAVS protein activity. Therefore, a novel peptide was synthesized to replace the function of the NLRX1 protein using a partial sequence of the NLRX1 protein at the binding site with the MAVS protein.

Example 2: Confirmation of antiviral reaction inhibitory efficacy of novel peptides

In order to confirm whether the novel peptides prepared in Example 1 are effective in inhibiting the antiviral response caused by virus infection, the following experiment was performed.

Specifically, in order to induce a viral infection response, a synthetic ribonucleic acid known to be associated with viral infection among pathogen-associated molecular types, promotes the production of interferon and an RNA virus gene analogue poly IC (polyinosinic: polycytidylic acid, polyIC) was used. First, a serum-free cell culture medium supplemented with poly IC 5 μg/ml and MQPX-79, MQPX-519, MQPX-751 or MQPX-779 was added to mouse lung epithelial cell line (MLE-12) 1 x 10 ^Five cells were treated for 16 hours. Thereafter, the supernatant was obtained, and the expression level of interferon-beta (IFN-β) was confirmed using an enzyme-linked immunosorbent assay (ELISA).

As a result, when MQPX-779 was treated among the novel peptides, it was confirmed that the expression level of IFN-β was significantly reduced compared to the control treated only with poly IC ( FIG. 2A ). In addition, as a result of performing the same experiment as above with various concentrations of MQPX-779 having excellent effect, it was confirmed that the expression level of IFN-β was significantly reduced even when 0.2 μM of MQPX-779 was treated ( FIG. 2B ). Based on the above results, it can be seen that MQPX-779 among the novel peptides prepared herein effectively inhibits the antiviral response caused by virus infection.

Example 3: Confirmation of cytotoxicity of novel peptides

In order to confirm whether the inflammation inhibitory activity of the novel peptides prepared in Example 1 is a result of the cytotoxicity of the peptide itself, the following experiment was performed.

Specifically, in order to confirm the cytotoxicity of the peptides, the XTT test method for measuring the activity of mitochondria in cells was used. First, a serum-free cell culture medium containing 5 μM and 50 μM of MQPX-79, MQPX-519, MQPX-751 or MQPX-779 was treated with a mouse peritoneal macrophage cell line (IC21) for 16 hours. . After that, XTT was treated for 1 hour and the amount of water-soluble formazan produced by reduction by dehydrogenase in mitochondria was measured.

As a result, it was confirmed that the cytotoxicity by the novel peptides of the present application does not appear in the concentration range used in the cell experiment ( FIG. 3 ), the inflammatory response inhibitory effect is not due to its own cytotoxicity, and the peptides It can be seen that even when used as a medicine, it is not harmful to the individual.

Example 4: Confirmation of Inflammasome Reaction Inhibition Efficacy of New Peptides

In order to confirm the effect of inhibiting the inflammasome reaction caused by bacterial infection of the novel peptides prepared in Example 1, the following experiment was performed.

Specifically, in order to induce an inflammasome response due to bacterial infection, LPS (lipopolysaccharide) and Gram-negative bacteria-derived outer membrane vesicle (OMV) were used as endotoxins. First, 5 µM of MQPX-79, MQPX-519, MQPX-751, or MQPX-779 was added to a serum-free cell culture medium containing 0.1 µg/ml of LPS or OMV to 1 x 10 ⁵ cells of a mouse peritoneal-derived macrophage for 6 hours. treated during Thereafter, the expression level of interleukin-1beta (IL-1β) in the obtained supernatant was confirmed by ELISA.

As a result, it was confirmed that the expression level of IL-1β was reduced when the novel peptides of the present application were treated, compared to the control treated only with LPS (FIG. 4A) or OMV (FIG. 4B). In particular, in the case of MQPX-779, other peptides It was confirmed that the expression level of IL-1β was reduced more significantly (FIG. 4). Based on the above results, it can be seen that the novel peptides prepared herein effectively inhibit the inflammasome reaction caused by bacterial infection.

Example 5: Confirmation of inflammatory response inhibitory efficacy of novel peptides

In order to confirm the inflammatory response inhibitory effect of the novel peptides prepared in Example 1, the following experiment was performed.

Specifically, to induce an inflammatory response caused by bacterial infection, LPS or OMV was used. First, 5 μM of MQPX-79, MQPX-519, MQPX-751, or MQPX-779 was added to a serum-free cell culture medium containing 0.1 μg/ml of LPS or OMV to 1 x 10 ⁵ cells of a mouse peritoneal-derived macrophage for 6 hours. treated during Thereafter, the expression levels of interleukin-6 (interleukin-6, IL-6) and tumor necrosis factor-alpha (TNF-α) in the obtained supernatant were confirmed by ELISA.

As a result, it was confirmed that MQPX-779 among the novel peptides of the present application significantly reduced the expression levels of IL-6 and TNF-α compared to the control treated with LPS or OMV only ( FIGS. 5 and 6 ). Based on the above results, it can be seen that the novel peptides prepared herein effectively inhibit the inflammatory response.

The foregoing description of the present application is for illustration, and those of ordinary skill in the art to which the present application pertains will understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present application. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. For example, each component described as a single type may be implemented in a dispersed form, and likewise components described as distributed may be implemented in a combined form.

The scope of the present application is indicated by the following claims rather than the above detailed description, and all changes or modifications derived from the meaning and scope of the claims and their equivalent concepts should be construed as being included in the scope of the present application.

<110> MITOQUEST <120> A novel anti-inflammatory peptides derived NLRX1 and use thereof <130> DP20200181KR <160> 5 <170> KoPatentIn 3.0 <210> 1 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> MQPX-779 <400> 1 Cys Gln Ile Thr Thr Leu Arg Leu Ser Asn Asn 1 5 10 <210> 2 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> MQPX-76 <400> 2 Glu Ala Ile Gln Arg His Arg Arg Asn Leu 1 5 10 <210> 3 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> MQPX-519 <400> 3 Arg Lys Thr Thr Leu Gln Arg Val Gly Lys 1 5 10 <210> 4 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> MQPX-751 <400> 4 Arg Ala Arg Lys Leu Gly Leu Gln Leu Asn 1 5 10 <210> 5 <211> 975 <212> PRT <213> Artificial Sequence <220> <223> NLRX1 <400> 5 Met Arg Trp Gly His His Leu Pro Arg Ala Ser Trp Gly Ser Gly Phe 1 5 10 15 Arg Arg Ala Leu Gln Arg Pro Asp Asp Arg Ile Pro Phe Leu Ile His 20 25 30 Trp Ser Trp Pro Leu Gln Gly Glu Arg Pro Phe Gly Pro Pro Arg Ala 35 40 45 Phe Ile Arg His His Gly Ser Ser Val Asp Ser Ala Pro Pro Pro Gly 50 55 60 Arg His Gly Arg Leu Phe Pro Ser Ala Ser Ala Thr Glu Ala Ile Gln 65 70 75 80 Arg His Arg Arg Asn Leu Ala Glu Trp Phe Ser Arg Leu Pro Arg Glu 85 90 95 Glu Arg Gln Phe Gly Pro Thr Phe Ala Leu Asp Thr Val His Val Asp 100 105 110 Pro Val Ile Arg Glu Ser Thr Pro Asp Glu Leu Leu Arg Pro Pro Ala 115 120 125 Glu Leu Ala Leu Glu His Gln Pro Pro Gln Ala Gly Leu Pro Pro Leu 130 135 140 Ala Leu Ser Gln Leu Phe Asn Pro Asp Ala Cys Gly Arg Arg Val Gln 145 150 155 160 Thr Val Val Leu Tyr Gly Thr Val Gly Thr Gly Lys Ser Thr Leu Val 165 170 175 Arg Lys Met Val Leu Asp Trp Cys Tyr Gly Arg Leu Pro Ala Phe Glu 180 185 190 Leu Leu Ile Pro Phe Ser Cys Glu Asp Leu Ser Ser Leu Gly Pro Ala 195 200 205 Pro Ala Ser Leu Cys Gln Leu Val Ala Gln Arg Tyr Thr Pro Leu Lys 210 215 220 Glu Val Leu Pro Leu Met Ala Ala Ala Gly Ser His Leu Leu Phe Val 225 230 235 240 Leu His Gly Leu Glu His Leu Asn Leu Asp Phe Arg Leu Ala Gly Thr 245 250 255 Gly Leu Cys Ser Asp Pro Glu Glu Pro Gln Glu Pro Ala Ala Ile Ile 260 265 270 Val Asn Leu Leu Arg Lys Tyr Met Leu Pro Gln Ala Ser Ile Leu Val 275 280 285 Thr Thr Arg Pro Ser Ala Ile Gly Arg Ile Pro Ser Lys Tyr Val Gly 290 295 300 Arg Tyr Gly Glu Ile Cys Gly Phe Ser Asp Thr Asn Leu Gln Lys Leu 305 310 315 320 Tyr Phe Gln Leu Arg Leu Asn Gln Pro Tyr Cys Gly Tyr Ala Val Gly 325 330 335 Gly Ser Gly Val Ser Ala Thr Pro Ala Gln Arg Asp His Leu Val Gln 340 345 350 Met Leu Ser Arg Asn Leu Glu Gly His His Gln Ile Ala Ala Ala Cys 355 360 365 Phe Leu Pro Ser Tyr Cys Trp Leu Val Cys Ala Thr Leu His Phe Leu 370 375 380 His Ala Pro Thr Pro Ala Gly Gln Thr Leu Thr Ser Ile Tyr Thr Ser 385 390 395 400 Phe Leu Arg Leu Asn Phe Ser Gly Glu Thr Leu Asp Ser Thr Asp Pro 405 410 415 Ser Asn Leu Ser Leu Met Ala Tyr Ala Ala Arg Thr Met Gly Lys Leu 420 425 430 Ala Tyr Glu Gly Val Ser Ser Arg Lys Thr Tyr Phe Ser Glu Glu Asp 435 440 445 Val Cys Gly Cys Leu Glu Ala Gly Ile Arg Thr Glu Glu Glu Phe Gln 450 455 460 Leu Leu His Ile Phe Arg Arg Asp Ala Leu Arg Phe Phe Leu Ala Pro 465 470 475 480 Cys Val Glu Pro Gly Arg Ala Gly Thr Phe Val Phe Thr Val Pro Ala 485 490 495 Met Gln Glu Tyr Leu Ala Ala Leu Tyr Ile Val Leu Gly Leu Arg Lys 500 505 510 Thr Thr Leu Gln Lys Val Gly Lys Glu Val Ala Glu Leu Val Gly Arg 515 520 525 Val Gly Glu Asp Val Ser Leu Val Leu Gly Ile Met Ala Lys Leu Leu 530 535 540 Pro Leu Arg Ala Leu Pro Leu Leu Phe Asn Leu Ile Lys Val Val Pro 545 550 555 560 Arg Val Phe Gly Arg Met Val Gly Lys Ser Arg Glu Ala Val Ala Gln 565 570 575 Ala Met Val Leu Glu Met Phe Arg Glu Glu Asp Tyr Tyr Asn Asp Asp 580 585 590 Val Leu Asp Gln Met Gly Ala Ser Ile Leu Gly Val Glu Gly Pro Arg 595 600 605 Arg His Pro Asp Glu Pro Pro Glu Asp Glu Val Phe Glu Leu Phe Pro 610 615 620 Met Phe Met Gly Gly Leu Leu Ser Ala His Asn Arg Ala Val Leu Ala 625 630 635 640 Gln Leu Gly Cys Pro Ile Lys Asn Leu Asp Ala Leu Glu Asn Ala Gln 645 650 655 Ala Ile Lys Lys Lys Leu Gly Lys Leu Gly Arg Gln Val Leu Pro Pro 660 665 670 Ser Glu Leu Leu Asp His Leu Phe Phe His Tyr Glu Phe Gln Asn Gln 675 680 685 Arg Phe Ser Ala Glu Val Leu Ser Ser Leu Arg Gln Leu Asn Leu Ala 690 695 700 Gly Val Arg Met Thr Pro Val Lys Cys Thr Val Val Ala Ala Val Leu 705 710 715 720 Gly Ser Gly Arg His Ala Leu Asp Glu Val Asn Leu Ala Ser Cys Gln 725 730 735 Leu Asp Pro Ala Gly Leu Arg Thr Leu Leu Pro Val Phe Leu Arg Ala 740 745 750 Arg Lys Leu Gly Leu Gln Leu Asn Ser Leu Gly Pro Glu Ala Cys Lys 755 760 765 Asp Leu Arg Asp Leu Leu Leu His Asp Gln Cys Gln Ile Thr Thr Leu 770 775 780 Arg Leu Ser Asn Asn Pro Leu Thr Ala Ala Gly Val Ala Val Leu Met 785 790 795 800 Glu Gly Leu Ala Gly Asn Thr Ser Val Thr His Leu Ser Leu Leu His 805 810 815 Thr Gly Leu Gly Asp Glu Gly Leu Glu Leu Leu Ala Ala Gln Leu Asp 820 825 830 Arg Asn Arg Gln Leu Gln Glu Leu Asn Val Ala Tyr Asn Gly Ala Gly 835 840 845 Asp Thr Ala Ala Leu Ala Leu Ala Arg Ala Ala Arg Glu His Pro Ser 850 855 860 Leu Glu Leu Leu His Leu Tyr Phe Asn Glu Leu Ser Ser Glu Gly Arg 865 870 875 880 Gln Val Leu Arg Asp Leu Gly Gly Ala Ala Glu Gly Gly Ala Arg Val 885 890 895 Val Val Ser Leu Thr Glu Gly Thr Ala Val Ser Glu Tyr Trp Ser Val 900 905 910 Ile Leu Ser Glu Val Gln Arg Asn Leu Asn Ser Trp Asp Arg Ala Arg 915 920 925 Val Gln Arg His Leu Glu Leu Leu Leu Arg Asp Leu Glu Asp Ser Arg 930 935 940 Gly Ala Thr Leu Asn Pro Trp Arg Lys Ala Gln Leu Leu Arg Val Glu 945 950 955 960 Gly Glu Val Arg Ala Leu Leu Glu Gln Leu Gly Ser Ser Gly Ser 965 970 975

Claims (10)

An anti-inflammatory peptide comprising at least one selected from the group consisting of amino acid sequences of SEQ ID NOs: 1 to 4 and modified amino acid sequences thereof.
According to claim 1,
The peptide comprises the amino acid sequence of SEQ ID NO: 1, anti-inflammatory peptide.
According to claim 1,
The peptide further comprises a cell-penetrating peptide, anti-inflammatory peptide.
According to claim 1,
The peptide is MAVS (mitochondrial antiviral-signaling) that inhibits aggregation or protein function, anti-inflammatory peptide.
According to claim 1,
The peptide is to inhibit the expression of inflammatory cytokines or inflammasome, anti-inflammatory peptide.
According to claim 1,
The peptide inhibits inflammation caused by bacteria or viruses, anti-inflammatory peptides.
Claims 1 to 6, which encodes the anti-inflammatory peptide of any one of claims, a polynucleotide.
An anti-inflammatory composition comprising the anti-inflammatory peptide of any one of claims 1 to 6 as an active ingredient.
A pharmaceutical composition for preventing or treating inflammatory diseases, comprising the anti-inflammatory peptide of any one of claims 1 to 6 as an active ingredient.
10. The method of claim 9,
The inflammatory diseases include sepsis, septic shock, systemic inflammatory response syndrome, acute respiratory distress syndrome, asthma, allergic and non-allergic rhinitis, chronic and acute rhinitis, chronic and Acute gastritis or enteritis, ulcerative gastritis, acute and chronic nephritis, acute and chronic hepatitis, chronic obstructive pulmonary disease, idiopathic pulmonary fibrosis (IPF) , irritable bowel syndrome, inflammatory pain, migraine, headache, back pain, fibromyalgia, myofascial disease, viral infection (such as hepatitis C infection), bacterial infection, fungal infection, burns, surgical or dental wounds, prostaglandin E excess syndrome, atherosclerosis, gout, arthritis, At least one selected from the group consisting of rheumatoid arthritis, ankylosing spondylitis, Hodgkin's disease, pancreatitis, conjunctivitis, iritis, scleritis, uveitis, dermatitis, atopic dermatitis, eczema, systemic lupus erythematosus (SLE) and multiple sclerosis Phosphorus, composition.

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