Hepatitis b virus-derived peptide, Anti-inflammatory use thereof, and pharmaceutical composition for preventing or treating inflammatory diseases comprising same

Abstract

The present invention relates to a hepatitis B virus-derived peptide, an anti-inflammatory use thereof, and a pharmaceutical composition for preventing or treating inflammatory diseases comprising same. A hepatitis B virus-derived peptide and a peptide in which an amino acid sequence is added, deleted, or substituted thereto, according to one aspect of the present invention, exhibit excellent anti-inflammatory activity and excellent safety, and a pharmaceutical composition comprising same may be effectively used for preventing or treating various inflammatory diseases, including an inflammatory bowel disease.

Description

Hepatitis B virus-derived peptide, anti-inflammatory use thereof, and pharmaceutical compositions containing the same for preventing or treating inflammatory diseases

[0001] The present invention relates to a hepatitis B virus-derived peptide, its anti-inflammatory use, and a pharmaceutical composition comprising the same for preventing or treating inflammatory diseases.

[0002] Inflammatory bowel disease (IBD) is a chronic inflammatory disease in which chronic inflammation occurs in the intestines, causing persistent symptoms such as diarrhea, bloody stool, weight loss, and abdominal pain. The underlying cause has not yet been clearly identified, and the symptoms persist throughout life. Even if the symptoms improve with medication, they are known to easily relapse. Therefore, it is an intractable disease that cannot be completely cured with modern medical technology and requires lifelong treatment and management.

[0003] Inflammatory bowel disease (IBD) was once considered a disease primarily prevalent in developed countries like North America and Europe. However, since the 2000s, its incidence has steadily increased in developing countries, including South America and Asia, amid rapid economic growth. According to the Global Burden of Disease (GBD) study, the number of patients with IBD has increased by nearly 85.1% worldwide since 1990, and the number of related deaths has increased by a staggering 67%.

[0004] Treatment for inflammatory bowel disease is divided into 'induction treatment', which aims to reduce clinical symptoms to a certain level, and 'maintenance treatment', which aims to prevent symptoms from recurring.

[0005] Among the currently used treatments, the drugs used for maintenance treatment are limited to aminosalicylic acid, TNF-α antibody treatment, and thiopurine due to side effects and efficacy issues. Among these, aminosalicylic acid has a low response rate and its effect is minimal in severe cases, so it is rarely used. In the case of thiopurine, more than 39% of patients discontinue taking it due to high toxicity and side effects.

[0006] Furthermore, several recent research papers have demonstrated a close link between the use of inflammatory bowel disease treatments and the risk of cancer. These treatments can also lead to serious side effects, such as immune-related side effects and an increased risk of infectious diseases, making long-term use difficult. Given the lifelong nature of inflammatory bowel disease, these issues have become significant limitations in its treatment.

[0007] In other words, current treatments for inflammatory bowel disease have various limitations, such as side effects, treatment refractoriness, and the risk of cancer development. Therefore, there is a need to develop a new treatment that reflects the unmet needs of inflammatory bowel disease, has fewer side effects, can be safely used for a long period of time, and maintains activity against the disease.

[0008] Meanwhile, the market for therapeutics using peptides is expected to reach $45.67 billion in 2023, growing steadily at an annual rate of 5.63% over the next 10 years, as mass production processes for peptides are established and their potential as therapeutics is re-evaluated.

[0009] Accordingly, the inventors of the present application sought to discover a novel peptide having a preventive and therapeutic effect on inflammatory bowel disease.

[0010] The inventors of the present application conducted in-depth research and elucidated the mechanism of action of the novel peptide, resulting in the discovery of a novel peptide derived from PreS1 of the hepatitis B virus that has excellent preventive and therapeutic effects not only on inflammatory bowel disease but also on various inflammatory diseases.

[0011] [Prior Art Literature]

[0012] [Patent Document]

[0013] (Patent Document 0001) Korean Patent Publication No. 10-2023-0033110

[0014] (Patent Document 0002) Korean Patent No. 10-2234027

[0015] One aspect of the present invention provides an isolated peptide comprising an amino acid sequence having at least 65% sequence homology or identity with the amino acid sequence of SEQ ID NO: 1, or a solvate or salt thereof.

[0016] Another aspect is to provide an isolated peptide comprising an amino acid sequence having at least 65% sequence homology or identity with the amino acid sequence of SEQ ID NO: 1, or a solvate or salt thereof.

[0017] Another aspect is to provide a pharmaceutical composition for preventing or treating an inflammatory disease, comprising the peptide, or a solvate or salt thereof, as an active ingredient.

[0018] Another aspect provides a method for preventing or treating an inflammatory disease comprising administering the peptide, or a solvate or salt thereof, to a subject in need thereof.

[0019] Another aspect provides the use of the peptide, or a solvate or salt thereof, for use in the manufacture of a pharmaceutical composition for the prevention or treatment of an inflammatory disease.

[0020] Another aspect provides the use of the peptide, or a solvate or salt thereof, for use in the prevention or treatment of an inflammatory disease.

[0021] Another aspect is to provide a health functional food for preventing or improving inflammatory diseases, comprising the peptide, or a solvate or salt thereof.

[0022] Another aspect provides the use of the peptide, or a solvate or salt thereof, for use in the manufacture of a health functional food for preventing or improving an inflammatory disease.

[0023] Another aspect provides the use of the peptide, or a solvate or salt thereof, for the prevention or amelioration of an inflammatory disease.

[0024] Another aspect is to provide a feed composition for preventing or improving an inflammatory disease, comprising the peptide, or a solvate or salt thereof.

[0025] Another aspect provides the use of the peptide, or a solvate or salt thereof, for use in the manufacture of a feed composition for preventing or improving an inflammatory disease.

[0026] One aspect of the present invention provides an isolated peptide comprising an amino acid sequence having at least 65% sequence homology or identity with the amino acid sequence of SEQ ID NO: 1, or a solvate or salt thereof.

[0027] A peptide according to one specific example of the present invention may be an amino acid sequence having a sequence homology or identity of at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the amino acids of SEQ ID NO: 1, or a peptide comprising the amino acid sequence of SEQ ID NO: 1.

[0028] Another aspect provides an isolated peptide comprising an amino acid sequence having at least 65% sequence homology or identity with the amino acid sequence of SEQ ID NO: 1, or a solvate or salt thereof.

[0029] A peptide according to one specific example of the present invention may be an amino acid sequence having a sequence homology or identity of at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% or more of the amino acid sequence of SEQ ID NO: 1, or a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

[0030] In one specific example, the peptide may be comprised of a sequence of 3 to 8, 4 to 8, or 4 to 7 amino acids. Specifically, the peptide may be comprised of a sequence of 4, 5, 6, or 7 amino acids.

[0031] In one specific example, the peptide may comprise the amino acid sequence of SEQ ID NO: 1; or comprise an amino acid sequence in which any one amino acid in the amino acid sequence of SEQ ID NO: 1 is substituted with another amino acid; or comprise an amino acid sequence in which one or two amino acid sequences are additionally linked to or deleted from the N-terminus of the amino acid sequence of SEQ ID NO: 1 or the substituted amino acid sequence; or comprise an amino acid sequence in which one or two amino acid sequences are additionally linked to or deleted from the C-terminus of the amino acid sequence of SEQ ID NO: 1 or the substituted amino acid sequence; or comprise an amino acid sequence in which one or two amino acid sequences are additionally linked to or deleted from the N-terminus of the amino acid sequence of SEQ ID NO: 1 or the substituted amino acid sequence, and one or two amino acid sequences are additionally linked to or deleted from the C-terminus.

[0032] In one specific example, the peptide may be composed of an amino acid sequence of SEQ ID NO: 1; or composed of an amino acid sequence in which any one amino acid in the amino acid sequence of SEQ ID NO: 1 is substituted with another amino acid; or composed of an amino acid sequence in which one or two amino acid sequences are additionally linked to or deleted from the N-terminus of the amino acid sequence of SEQ ID NO: 1 or the substituted amino acid sequence; or composed of an amino acid sequence in which one or two amino acid sequences are additionally linked to or deleted from the C-terminus of the amino acid sequence of SEQ ID NO: 1 or the substituted amino acid sequence; or composed of an amino acid sequence in which one or two amino acid sequences are additionally linked to or deleted from the N-terminus of the amino acid sequence of SEQ ID NO: 1 or the substituted amino acid sequence, and one or two amino acid sequences are additionally linked to or deleted from the C-terminus.

[0033] In one specific example, the peptide may comprise the amino acid sequence of SEQ ID NO: 1; or comprise an amino acid sequence in which any one amino acid in the amino acid sequence of SEQ ID NO: 1 is substituted with another amino acid; or comprise an amino acid sequence in which one or two amino acid sequences are deleted at the N-terminus of the amino acid sequence of SEQ ID NO: 1 or the substituted amino acid sequence; or comprise an amino acid sequence in which a methionine is additionally bonded to the N-terminus of the amino acid sequence of SEQ ID NO: 1 or the substituted amino acid sequence; or comprise an amino acid sequence in which a methionine is additionally bonded to the N-terminus of the amino acid sequence of SEQ ID NO: 1 or the substituted amino acid sequence and one or two amino acid sequences are deleted at the C-terminus.

[0034] In one specific example, the peptide may be composed of an amino acid sequence of SEQ ID NO: 1; or composed of an amino acid sequence in which any one amino acid in the amino acid sequence of SEQ ID NO: 1 is substituted with another amino acid; or composed of an amino acid sequence in which one or two amino acid sequences are deleted from the N-terminus of the amino acid sequence of SEQ ID NO: 1 or the substituted amino acid sequence; or composed of an amino acid sequence in which a methionine is additionally bonded to the N-terminus of the amino acid sequence of SEQ ID NO: 1 or the substituted amino acid sequence; or composed of an amino acid sequence in which a methionine is additionally bonded to the N-terminus of the amino acid sequence of SEQ ID NO: 1 or the substituted amino acid sequence and one or two amino acid sequences are deleted from the C-terminus.

[0035] In one specific example, the peptide may comprise an amino acid sequence of any one of the sequences of SEQ ID NOs: 1 to 6.

[0036] In one specific example, the peptide may be composed of any one of the amino acid sequences of SEQ ID NOs: 1 to 6.

[0037] As used herein, the term "peptide" may mean a linear molecule formed by amino acid residues being linked to each other by peptide bonds.

[0038] The above peptide can be obtained by various peptide synthesis methods widely known in the art. For example, it can be produced using polynucleotide recombination and protein expression systems, chemical synthesis methods (e.g., solid-phase synthesis or liquid-phase synthesis), cell-free protein synthesis, etc. In addition, as an example, the peptide can be a product obtained by culturing a peptide, an extract of plant-derived tissue or cells, or a microorganism (e.g., bacteria or fungi, and particularly yeast), and specifically, it can be derived from PreS1 of the hepatitis B virus (HBV).

[0039] The above amino acid may be any one selected from the group consisting of alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, tryptophan, serine, threonine, asparagine, glutamine, cysteine, glycine, proline, arginine, histidine, lysine, aspartic acid, glutamic acid and all variants of the above amino acids.

[0040] The above substitution may be a conservative substitution.

[0041] The above substitution may be a substitution of an amino acid corresponding to position 1, position 2, position 3, position 4, position 5, or position 6 in the amino acid sequence of sequence number 1 with another amino acid.

[0042] As used herein, the term "conservative substitution" refers to the replacement of an amino acid residue with an amino acid residue having a similar side chain, without causing a loss of biological or biochemical function of the peptide or protein. Classes of amino acid residues having similar side chains are well known and defined in the art. These classes include amino acids with basic side chains (e.g., lysine (K), arginine (R), histidine (H)), amino acids with acidic side chains (e.g., aspartic acid (D), glutamic acid (E)), amino acids with uncharged polar side chains (e.g., glycine, asparagine (N), glutamine (Q), serine (S), threonine (T), tyrosine (Y), cysteine ​​(C)), amino acids with nonpolar side chains (e.g., alanine (A), valine (V), leucine (L), isoleucine (I), proline (P), phenylalanine (F), methionine (M), tryptophan (W)), amino acids with beta-branched side chains (e.g., threonine (T), valine (V), isoleucine (I)) and amino acids with aromatic side chains (e.g., tyrosine (Y), phenylalanine (F), tryptophan (W), histidine (H). That is, the peptide may have, for example, one or more conservative substitutions while still retaining the biological activity of the intact peptide. Such amino acid substitutions may generally occur based on similarities in the polarity, charge, solubility, hydrophobicity, hydrophilicity, and / or amphipathic nature of the residues. For example, positively charged (basic) amino acids include arginine, lysine, and histidine; negatively charged (acidic) amino acids include glutamic acid and aspartic acid; aromatic amino acids include phenylalanine, tryptophan, and tyrosine; and hydrophobic amino acids include alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan.Additionally, amino acids can be classified into those with electrically charged side chains and those with uncharged side chains. Amino acids with electrically charged side chains include aspartic acid, glutamic acid, lysine, arginine, and histidine. Amino acids with uncharged side chains can be further classified into nonpolar amino acids or polar amino acids. Nonpolar amino acids include glycine, alanine, valine, leucine, and isoleucine. methionine, and proline, and polar amino acids include serine, threonine, cysteine, asparagine, and glutamine. Conservative substitutions with amino acids having similar properties as described above can be expected to exhibit the same or similar activities.

[0043] As used herein, the terms "amino acid" and "amino acid residue" refer to a natural amino acid, an unnatural amino acid, or a modified amino acid. Unless otherwise stated, all references to amino acids, either generically or by name, specifically include references to both the D and L stereoisomers (where the structure permits such stereoisomeric forms). Natural amino acids include alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine ​​(Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), and valine (Val). Non-natural amino acids include modified amino acid residues that are chemically modified, or reversibly or irreversibly chemically blocked, at the N-terminal amino group or side chain, such as N-methylated D and L amino acids or residues in which a side chain functional group is chemically modified with another functional group.

[0044] Abbreviations are used to describe amino acids in this document.

[0045] Table 1 below lists amino acid abbreviations.

[0046] Amino Acids 3-letter Abbreviations 1-letter Abbreviations Alanine Ala AA Valine Val VIsoleucine Ile LEucine Leu L Methionine Met M Phenylalanine Phe F Tyrosine Tyr Y Tryptophan Trp W Serine Ser S Threonine Thr T Asparagine Asn NGlutamine Gln Q Cysteine ​​Cys C Glycine Gly GProline Pro PArginine Arg RHistidine His HLysine Lys K Aspartic Acid Asp D Glutamic Acid Glu E

[0047]

[0048] Another aspect provides a nucleic acid molecule comprising a nucleotide sequence encoding the peptide according to the present invention, or a vector comprising the nucleic acid molecule.

[0049] The term "vector" as used herein may refer to a polynucleotide product containing a regulatory sequence and a base sequence of a gene so as to enable expression of the gene of interest in a suitable host cell.

[0050] It is obvious that a polynucleotide having a peptide or nucleotide sequence having an amino acid sequence having homology or identity described herein and exhibiting an activity corresponding to the peptide or polynucleotide described herein, in which a part of the sequence is deleted, modified, substituted (e.g., conservatively substituted), or added, is also included within the scope of the present invention.

[0051] As used herein, the terms "identity" or "homology" refer to the degree of resemblance between two given amino acid sequences or nucleotide sequences, which may be expressed as a percentage. The terms identity and homology may be used interchangeably.

[0052] Sequence identity or homology of peptides or polynucleotides is determined by standard alignment algorithms, and may be combined with default gap penalties established by the program being used. In practice, sequences that are homologous or identical are generally capable of hybridizing under moderate or high stringency conditions along at least about 50%, 60%, 70%, 80%, or 90% of the entire sequence or its full length.

[0053] Whether any two peptide or polynucleotide sequences have homology, similarity or identity can be determined using, for example, but not limited to, BLAST of the National Center for Biotechnology Information Database or ClustalW.

[0054] Additionally, whether any two peptide or polynucleotide sequences have homology, similarity or identity can be determined by comparing the sequences by Southern hybridization experiments under defined stringent conditions, and the appropriate hybridization conditions defined are within the scope of the art and can be determined by methods well known to those skilled in the art.

[0055] In the present invention, the “corresponding amino acid” refers to an amino acid residue at a corresponding position in a peptide, or an amino acid residue that is similar, identical, or homologous to the amino acid residue at the corresponding position. Identifying the amino acid at the corresponding position may determine a specific amino acid of a sequence that references a specific sequence. In the present invention, the “corresponding position” generally refers to a similar or corresponding position in the amino acid sequence of a related protein or a reference sequence. For example, any amino acid sequence may be aligned with SEQ ID NO: 1, and based on this, each amino acid residue of the amino acid sequence may be numbered by referring to the numerical position of the amino acid residue corresponding to the amino acid residue in SEQ ID NO: 1. For example, the position of the corresponding amino acid, or the position where a modification such as a substitution, addition, or deletion occurs, can be identified by comparing it with a query sequence (also referred to as a “reference sequence”) using a sequence alignment algorithm known in the art.

[0056] It is clear that a peptide of the present invention has an amino acid sequence that has the identity or homology described herein and exhibits an activity corresponding to the peptide, and a peptide having an amino acid sequence in which a portion of the sequence is deleted, modified, substituted or added is also included within the scope of the present invention.

[0057] Specifically, the peptides of the present invention may include deletions or additions of amino acids that have minimal impact on the properties and secondary structure of the peptide, and for example, the N-terminus of the variant may be conjugated with a signal (or leader) sequence that is involved in translocation of the protein co-translationally or post-translationally.

[0058] Additionally, the peptides of the present invention may be conjugated with other sequences or linkers to enable identification, purification, or synthesis.

[0059] In addition, if it has the same or corresponding activity as the peptide of the present invention, it does not exclude meaningless sequence additions before and after the amino acid sequence of the peptide, mutations that may occur naturally, or silent mutations thereof, and it is clear that even if it has such sequence additions or mutations, it falls within the scope of the present invention.

[0060] As used herein, the term "solvate" refers to a molecular complex between a peptide according to the present invention and solvent molecules, and may refer to a peptide according to the present invention or a salt thereof containing a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces. Examples of solvates include, but are not limited to, a peptide according to the present invention bound to water, isopropanol, ethanol, methanol, dimethylsulfoxide, ethyl acetate, acetic acid, ethanolamine, or a mixed solvent thereof.

[0061] Additionally, it may be convenient or desirable to prepare, purify, and / or handle a corresponding solvate of the peptide. The term "solvate" may be used herein in its conventional sense to refer to a complex of a solute (e.g., a peptide, a salt of the peptide) and a solvent. When the solvent is water, the solvate may conveniently be referred to as a hydrate, such as a monohydrate, dihydrate, trihydrate, etc.

[0062] In one specific example, the peptide may be derived from hepatitis B virus (HBV) PreS1.

[0063] In one specific example, the peptide may have anti-inflammatory activity.

[0064] In one specific example, the peptide may have anti-inflammatory activity by inducing the expression of CXCL9 in macrophages and attracting CXCR3+ T cells.

[0065] According to one embodiment of the present invention, a peptide may have a protecting group attached to the N- or C-terminus of the peptide to obtain chemical stability, enhanced pharmacological properties (half-life, absorbability, potency, efficacy, etc.), altered specificity (e.g., a broad spectrum of biological activity), and reduced antigenicity. Any component capable of modifying the peptide, particularly enhancing the stability of the peptide, may be included without limitation.

[0066] As used herein, the term "stability" may mean not only in vivo stability, which protects the peptide from attack by in vivo protein cleavage enzymes, but also storage stability (e.g., room temperature storage stability).

[0067] In one specific example, the N-terminus of the peptide may be bound to any one protecting group selected from the group consisting of an acetyl group, a fluoreonylmethoxycarbonyl group, a formyl group, a palmitoyl group, a myristyl group, a stearyl group, a butoxycarbonyl group, an aryloxycarbonyl group, and polyethylene glycol (PEG).

[0068] In one specific example, the C-terminus of the peptide may be bound to any one protecting group selected from the group consisting of an amino group (-NH2), a tertiary alkyl group, and an azide (-NHNH2).

[0069] Additionally, the peptide may additionally include an amino acid sequence manufactured for a specific purpose, such as a targeting sequence, a tag, or a labeled residue.

[0070]

[0071] Another aspect provides a pharmaceutical composition for preventing or treating an inflammatory disease, comprising the peptide according to the present invention, or a solvate or salt thereof, as an active ingredient.

[0072] Another aspect provides a method for preventing or treating an inflammatory disease, comprising administering to a subject in need thereof the peptide according to the present invention, or a solvate or salt thereof.

[0073] Another aspect provides the use of the peptide according to the present invention, or a solvate or salt thereof, for use in the manufacture of a pharmaceutical composition for the prevention or treatment of an inflammatory disease.

[0074] Another aspect provides the use of the peptide according to the present invention, or a solvate or salt thereof, for the prevention or treatment of an inflammatory disease.

[0075] In one specific example, the inflammatory disease is inflammatory bowel disease, irritable bowel syndrome, sepsis, septic shock, chronic inflammatory disease caused by chronic viral or bacterial infection, pancreatitis, gastric ulcer, gastritis, rhinitis, bronchitis, periodontitis, inflammatory skin disease, atopic dermatitis, encephalitis, chronic obstructive pulmonary disease, pulmonary fibrosis, undifferentiated spondyloarthropathy, undifferentiated arthropathy, arthritis, inflammatory osteolysis, inflammatory collagen vascular disease, nephritis, glomerulonephritis, type 1 diabetes, rheumatoid arthritis, reactive arthritis, osteoarthritis, psoriasis, scleroderma, osteoporosis, atherosclerosis, myocarditis, endocarditis, pericarditis, cystic fibrosis, Hashimoto's thyroiditis, Graves' disease, leprosy, syphilis, Lyme disease, borreliosis, neurogenic borreliosis, tuberculosis, sarcoidosis, lupus, discoid It may be one or more selected from the group consisting of lupus, chilblain lupus, lupus nephritis, systemic lupus erythematosus, macular degeneration, uveitis, Crohn's disease, Sjogren's syndrome, fibromyalgia, chronic fatigue syndrome, chronic fatigue immunodeficiency syndrome, myalgic encephalomyelitis, amyotrophic lateral sclerosis, Parkinson's disease, and multiple sclerosis.

[0076] In one specific example, the inflammatory bowel disease may be at least one selected from the group consisting of Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, ischemic colitis, diversionary colitis, colitis, hemorrhagic rectal ulcer, Behcet's disease, inflammatory enteritis, chronic enteritis, acute enteritis, intestinal tuberculosis, ileal pouchitis, bacterial enteritis, viral enteritis, amoebic enteritis, and ischemic colitis.

[0077] In a pharmaceutical composition according to one specific example, the peptide may have anti-inflammatory activity.

[0078] In a pharmaceutical composition according to one specific example, the peptide may have anti-inflammatory activity by inducing the expression of CXCL9 in macrophages and attracting CXCR3+ T cells.

[0079] In a pharmaceutical composition according to one embodiment, the peptide may have a protecting group attached to the N- or C-terminus of the peptide to obtain chemical stability, enhanced pharmacological properties (half-life, absorbability, potency, efficacy, etc.), altered specificity (e.g., a broad spectrum of biological activity), and reduced antigenicity. Any component capable of modifying the peptide, particularly enhancing the stability of the peptide, may be included without limitation.

[0080] In a pharmaceutical composition according to one specific example, the N-terminus of the peptide may be bound to any one protecting group selected from the group consisting of an acetyl group, a fluoreonylmethoxycarbonyl group, a formyl group, a palmitoyl group, a myristyl group, a stearyl group, a butoxycarbonyl group, an aryloxycarbonyl group, and polyethylene glycol (PEG).

[0081] In a pharmaceutical composition according to one specific example, the C-terminus of the peptide may be bound to any one protecting group selected from the group consisting of an amino group (-NH2), a tertiary alkyl group, and an azide (-NHNH2).

[0082] The term "comprising as an active ingredient" in this specification means including an effective amount capable of exhibiting the preventive or therapeutic effect of the pharmaceutical composition on an inflammatory disease.

[0083] In a pharmaceutical composition according to one embodiment, the salt may be a pharmaceutically acceptable salt.

[0084] As used herein, the term “pharmaceutically acceptable” means a substance that can be effectively used for a desired purpose without causing excessive toxicity, irritation, or allergic reaction, within the scope of pharmaceutical judgment.

[0085] As used herein, the term "pharmaceutically acceptable salt" means a salt according to one aspect of the present invention that is pharmaceutically acceptable and has the desired pharmacological activity of the parent compound (e.g., a peptide). Salts of the parent compound can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid form of these compounds with a stoichiometric amount of a suitable base, for example, sodium, calcium, magnesium or potassium, or by reacting the free base form of these compounds with a stoichiometric amount of a suitable acid. These reactions are typically carried out in water or in an organic solvent or in a mixture of the two. Generally, when practical, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile can be used. The pharmaceutically acceptable salts include both addition salts of acids or bases and stereochemically isomeric forms thereof, and may be, for example, addition salts of organic or inorganic acids. The above salt includes any salt that maintains the activity of the parent compound in the subject of administration and does not cause undesirable effects, and is not particularly limited thereto.

[0086] These salts include inorganic and organic salts, for example, acetic acid, nitric acid, aspartic acid, sulfonic acid, sulfuric acid, maleic acid, glutamic acid, formic acid, succinic acid, phosphoric acid, phthalic acid, tannic acid, tartaric acid, hydrobromic acid, propionic acid, benzenesulfonic acid, benzoic acid, stearic acid, lactic acid, bicarboxylic acid, bisulfuric acid, bitartaric acid, oxalic acid, butyric acid, calcium idet, carbonic acid, chlorobenzoic acid, citric acid, idetic acid, toluenesulfonic acid, fumaric acid, gluceptic acid, esilinic acid, pamoic acid, gluconic acid, methylnitric acid, malonic acid, hydrochloric acid, hydroiodoic acid, hydroxynaphtholic acid, isethionic acid, lactobionic acid, mandelic acid, mucic acid, It can be naphthylic acid, muconic acid, p-nitromethanesulfonic acid, hexamic acid, pantothenic acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, salicylic acid, sulfamic acid, sulfanilinic acid, methanesulfonic acid. In addition, the salt form includes salts of alkali and alkaline earth metals such as ammonium salt, lithium salt, sodium salt, potassium salt, magnesium salt, and calcium salt, salts with organic bases such as benzathine, N-methyl-D-glucamine, and hydrabamine salts, and salts with amino acids such as arginine and lysine. In addition, the salt form can be converted into a free form by treating with a suitable base or acid.

[0087] In this specification, the term “prevention” means any action that inhibits or delays the onset of a target disease, “treatment” means any action that improves or beneficially changes a target disease and its metabolic abnormality symptoms by administering a pharmaceutical composition according to the present invention, and “improvement” means any action that reduces a parameter related to a target disease, for example, the severity of a symptom, by administering a composition according to the present invention.

[0088] In one specific example, the pharmaceutical composition may be administered as a separate therapeutic agent or may be administered in combination with an anti-inflammatory agent or anti-inflammatory agent. The anti-inflammatory agent or anti-inflammatory agent refers to a conventional anti-inflammatory agent or anti-inflammatory agent for the prevention or treatment of inflammatory diseases.

[0089] As used herein, the term “combination administration” may be achieved by administering the individual components of the treatment regimen simultaneously, sequentially, in reverse order, or separately.

[0090] The terms "subject" and "patient" are used interchangeably herein. The subject may be an animal. In some embodiments, the subject is a mammal, such as a non-human animal (e.g., a cow, pig, horse, cat, dog, rat, mouse, monkey, or other primate). In some embodiments, the subject is a cynomolgus monkey. In some embodiments, the subject is a human.

[0091] The above pharmaceutical composition is administered in a therapeutically effective amount.

[0092] As used herein, the term "therapeutically effective amount" means an amount of a drug, e.g., the peptide according to the present invention, or a solvate or salt thereof, effective to achieve a desired therapeutic or prophylactic result. In some cases, the desired result is treatment of a disease or disorder in a subject. The therapeutically effective amount level may be determined based on factors including the type and severity of the patient's disease, the activity and sensitivity of the drug to the drug, the time of administration, the route of administration and excretion rate, the duration of treatment, concomitant drugs, and other factors well known in the medical field. The compositions of the present disclosure may be administered as individual therapeutic agents or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or in multiple doses. That is, the total effective amount of the compositions of the present disclosure may be administered to a patient as a single dose, or may be administered by a fractionated treatment protocol in which multiple doses are administered over a long period of time. Taking all of the above factors into consideration, it is important to administer the amount that can achieve the maximum effect with the minimum amount without side effects, and this can be easily determined by those skilled in the art.

[0093] As used herein, terms such as "treating," "treatment," "to treat," "palliating," or "to palliate" refer to therapeutic measures aimed at curing, slowing, alleviating symptoms, and / or arresting the progression of a diagnosed pathological condition or disorder. Therefore, those requiring treatment include those who have already been diagnosed with or are suspected of having a disorder.

[0094] Meanwhile, the pharmaceutical composition according to the present invention may additionally include a pharmaceutically acceptable carrier and may be formulated together with the carrier.

[0095] As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier or diluent that does not stimulate a living organism and does not inhibit the biological activity and properties of the administered compound. In a composition formulated as a liquid solution, acceptable pharmaceutical carriers include those that are sterile and biocompatible, such as saline solution, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and a mixture of one or more of these components. If necessary, other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate the composition into injectable formulations such as aqueous solutions, suspensions, and emulsions, pills, capsules, granules, or tablets.

[0096] The pharmaceutical composition according to one embodiment of the present invention and the composition comprising a pharmaceutically acceptable carrier can be applied to any dosage form containing the pharmaceutical composition as an active ingredient, and can be prepared as an oral or parenteral dosage form, and can be formulated in a unit dosage form for ease of administration and uniformity of dosage. The pharmaceutical dosage form of the present specification includes a form suitable for oral, rectal, nasal, topical (including buccal and sublingual), subcutaneous, vaginal, or parenteral (including intramuscular, subcutaneous, and intravenous) administration, or a form suitable for administration by inhalation or insufflation.

[0097] Oral administration dosage forms containing the composition of the present invention as an active ingredient may be formulated, for example, as tablets, troches, lozenges, aqueous or oily suspensions, prepared powders or granules, emulsions, hard or soft capsules, syrups or elixirs.

[0098] The composition of the present invention may be formulated as a parenteral dosage form containing the active ingredient, such as a subcutaneous injection, intravenous injection, or intramuscular injection; a suppository injection; or a spray, such as an aerosol, that can be inhaled through the respiratory tract. To formulate the composition of the present invention as an injectable dosage form, the composition of the present invention may be mixed with a stabilizer or buffer in water to prepare a solution or suspension, which may then be formulated into a unit dosage form in an ampoule or vial.

[0099] The dosage of the pharmaceutical composition of the present invention varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and disease severity. The daily dosage is preferably 0.01 μg to 1000 mg per kg of body weight per day when administered parenterally, more preferably 1 μg to 500 mg. However, since the dosage may increase or decrease depending on the route of administration, severity of obesity, sex, body weight, age, etc., the above dosage does not limit the scope of the present invention in any way.

[0100]

[0101] Another aspect provides a health functional food for preventing or improving inflammatory diseases, comprising the peptide according to the present invention, or a solvate or salt thereof.

[0102] Another aspect provides the use of the peptide according to the present invention, or a solvate or salt thereof, for use in the manufacture of a health functional food for preventing or improving inflammatory diseases.

[0103] Another aspect provides the use of the peptide according to the present invention, or a solvate or salt thereof, for the prevention or improvement of inflammatory diseases.

[0104] In the health functional food of the present invention according to one specific example, “peptide”, “solvent”, “inflammatory disease”, “prevention” and “improvement” are as described above.

[0105] In the health functional food of the present invention according to one specific example, the salt may be a salt acceptable from a food science perspective.

[0106] As used herein, the term "food-acceptable salt" refers to a formulation of a compound (e.g., a peptide) that does not cause serious irritation to an organism to which the compound is administered and does not impair the biological activity and physical properties of the compound. For example, the food-acceptable salt can be obtained by reacting the compound with an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, sulfonic acid such as methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, tartaric acid, formic acid, citric acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, capric acid, isobutanoic acid, malonic acid, succinic acid, phthalic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, salicylic acid, and the like. In addition, it can be obtained by reacting the compound with a base to form a salt such as an alkali metal salt such as an ammonium salt, a sodium or potassium salt, an alkaline earth metal salt such as a calcium or magnesium salt, a salt of an organic base such as dicyclohexylamine, N-methyl-D-glucamine, tris(hydroxymethyl)methylamine, etc., but is not limited thereto.

[0107] The health functional food herein may be formulated into any one form selected from the group consisting of powders, tablets, capsules, pills, granules, and liquids, using conventional methods known in the art, but is not limited thereto. Various forms may be manufactured using methods known in the art.

[0108] Additionally, it can be prepared in the form of a composition by mixing with a known substance or active ingredient known to have an activity of preventing, improving, or treating inflammatory diseases.

[0109] In addition, the health functional food of the present invention may contain conventional food additives, and the suitability as the "food additive" is determined by the specifications and standards for the relevant item according to the general provisions and general test methods of the Food Additives Codex approved by the Ministry of Food and Drug Safety, unless otherwise specified. Items listed in the "Food Additives Codex" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon pigment, licorice extract, crystalline cellulose, sorghum pigment, and guar gum; and mixed preparations such as sodium L-glutamate preparations, alkaline agents added to noodles, preservative preparations, and tar color preparations.

[0110] In addition to the above, the health functional food of the present invention may include various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the composition of the present invention may include fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These ingredients may be used independently or in combination.

[0111]

[0112] Another aspect provides a feed composition for preventing or improving an inflammatory disease comprising the peptide according to the present invention, or a solvate or salt thereof.

[0113] Another aspect provides the use of the peptide according to the present invention, or a solvate or salt thereof, for use in the preparation of a feed composition for preventing or improving an inflammatory disease.

[0114] Another aspect provides the use of the peptide according to the present invention, or a solvate or salt thereof, for the prevention or improvement of inflammatory diseases.

[0115] In the feed composition of the present invention according to one specific example, “peptide”, “solvent”, “inflammatory disease”, “prevention” and “improvement” are as described above.

[0116] In this specification, “feed composition” may mean any natural or artificial diet, meal, etc. or a component of said meal, especially for or suitable for eating, ingesting and digesting by an animal.

[0117] The above feed composition may include, but is not particularly limited to, nutrients such as energy, protein, lipids, vitamins, and minerals required by the individual consuming the feed. The individual refers to the subject of breeding, and includes, without limitation, any living organism capable of consuming the feed of the present invention, including companion animals and livestock.

[0118] The type of the above feed is not particularly limited, and feed commonly used in the relevant technical field can be used. Non-limiting examples of the above feed include plant-based feed such as grains, roots, food processing by-products, algae, fiber, pharmaceutical by-products, oils, starches, meal, or grain by-products; and animal-based feed such as proteins, inorganic substances, oils, minerals, oils, single-cell proteins, zooplankton, or food. These may be used alone or in combination of two or more types.

[0119] The feed composition according to the present invention can be manufactured by adding the peptide of the present invention, or a solvate or salt thereof, in an appropriate effective concentration range according to various feed manufacturing methods known in the art.

[0120] The feed composition according to the present invention can be applied without limitation to any individual for the purpose of preventing or improving inflammatory diseases. For example, it can be applied to any individual, including non-human animals such as monkeys, dogs, cats, rabbits, guinea pigs, rats, mice, cows, sheep, pigs, goats, birds, and fish.

[0121] A hepatitis B virus-derived peptide according to one aspect of the present invention and a peptide having an amino acid sequence added, deleted or substituted thereto have excellent anti-inflammatory activity and excellent safety, and a pharmaceutical composition comprising the same can be usefully utilized for the prevention or treatment of various inflammatory diseases, including inflammatory bowel disease.

[0122] Figure 1 is a schematic diagram showing a hepatitis B virus-derived peptide (Up7del1).

[0123] Figure 2 shows a schematic diagram of the evaluation of Up7del1 efficacy using a DSS-induced colitis model.

[0124] Figure 3 shows the method of inducing acute colitis through DSS.

[0125] Figure 4 shows the method of inducing chronic colitis through DSS.

[0126] Figure 5 is a graph evaluating the anti-inflammatory efficacy of Up7del1 in an inflammation-induced model using W4P-LHB-NIH3T3.

[0127] Figure 6 is a graph evaluating the anti-inflammatory efficacy of novel peptides “Up7del1, Up7del2, Up7del3, Up5, Up6, and Up7” in an inflammation-induced model using W4P-LHB-NIH3T3.

[0128] Figure 7 is a graph evaluating the anti-inflammatory efficacy of Up7del1 in an in vivo sepsis model.

[0129] Figure 8 is a graph evaluating the cytotoxicity of Up7del1 using the Neutral red uptake (NRU) assay.

[0130] Figure 9 is a summary table of the results of a biotoxicity test upon single subcutaneous administration of Up7del1.

[0131] Figure 10 is a summary table of the results of a biotoxicity test upon repeated subcutaneous administration of Up7del1.

[0132] Figure 11 is a photograph and graph evaluating the level of relief of DSS-induced acute colitis by Up7del1. In Figure 11, item A represents weight change, item B represents change in disease activity, item C represents comparison of colon length, and item D represents histological evaluation within the colon.

[0133] Figure 12 is a graph evaluating the survival rate by Up7del1 in a DSS-induced acute colitis model.

[0134] Figure 13 is a graph confirming the optimal Up7del1 dosage in a DSS-induced acute colitis model. In Figure 13, item A represents colon length comparison, item B represents weight change, and item C represents change in disease activity.

[0135] Figure 14 is a photograph and graph evaluating the efficacy of Up7del1 in a DSS-induced chronic colitis model. In Figure 14, item A represents colon length comparison, item B represents weight change, and item C represents change in disease activity.

[0136] Figure 15 shows changes in blood inflammatory cytokines caused by Up7del1 in a DSS-induced acute colitis model.

[0137] Figure 16 shows changes in the expression of inflammatory cytokines in colon cells by Up7del1 in a DSS-induced acute colitis model.

[0138] Figure 17 shows the change in the proportion of Th17 and Treg in the colon by Up7del1 in the DSS-induced acute colitis model.

[0139] Figure 18 shows the change in the proportion of Th17 and Treg in the mesenteric lymph nodes by Up7del1 in the DSS-induced acute colitis model.

[0140] Figure 19 shows changes in the differentiation pattern of colonic macrophages by Up7del1 in a DSS-induced acute colitis model.

[0141] Figure 20 shows changes in the differentiation pattern of macrophages in lymph nodes by Up7del1 in a DSS-induced acute colitis model.

[0142] Figure 21 shows changes in the expression pattern of colonic macrophages by Up7del1 in a DSS-induced acute colitis model.

[0143] Figure 22 shows an increase in CXCL9+ macrophages and CXCR3+ regulatory T cells in the colon following Up7del1 treatment as confirmed by confocal microscopy.

[0144] Figure 23 shows an increase in CXCL9+ macrophages and CXCR3+ regulatory T cells in the colon following Up7del1 treatment as confirmed by flow cytometry.

[0145] Figure 24 shows the experimental schedule and administration schedule to confirm the decrease in the therapeutic efficacy of Up7del1 on DSS-induced acute colitis when CXCL9 is blocked.

[0146] Figure 25 is a graph evaluating the decrease in the efficacy of Up7del1 in treating DSS-induced acute colitis by CXCL9 blocking through changes in body weight.

[0147] Figure 26 is a graph evaluating the decrease in the therapeutic efficacy of Up7del1 in DSS-induced acute colitis by CXCL9 blocking through changes in disease activity.

[0148] Figure 27 is a photograph and graph evaluating the reduction in the efficacy of Up7del1 in treating DSS-induced acute colitis by CXCL9 blocking through comparison of colon length.

[0149] Figure 28 is a graph histologically evaluating the decrease in the efficacy of Up7del1 in treating DSS-induced acute colitis when CXCL9 is blocked.

[0150] Figures 29 to 32 are graphs evaluating the reduction in anti-inflammatory efficacy of Up7del1 in DSS-induced acute colitis when CXCL9 is blocked.

[0151] Hereinafter, preferred embodiments are presented to aid understanding of the present invention. However, the following embodiments are provided solely to facilitate a better understanding of the present invention and are not intended to limit the scope of the present invention. The embodiments are susceptible to various modifications, and thus the embodiments are not limited to the embodiments disclosed below and may be implemented in various forms.

[0152] Terms or words used in the specification and claims of the present invention are not to be construed as limited to their usual or dictionary meanings, and should be interpreted as meanings and concepts that conform to the technical idea of ​​the present invention based on the principle that the inventor can appropriately define the concept of the term to explain his or her own invention in the best way.

[0153] Throughout the specification of the present invention, when a part is said to "include" a certain component, this does not mean that other components are excluded, but rather that other components may be included, unless specifically stated otherwise.

[0154] Throughout the specification of the present invention, “A and / or B” means A or B, or A and B.

[0155]

[0156] The research results of this patented invention are the result of research (RS-2025-00553721) conducted with support from the National Research Foundation of Korea with funding from the government (Ministry of Science and ICT).

[0157]

[0158] Example 1. Discovery of a novel peptide with anti-inflammatory properties.

[0159] The present invention was designed to determine whether a peptide derived from hepatitis B virus (HBV) has anti-inflammatory effects.

[0160] Specifically, a library of various peptides derived from the PreS1 protein sequence of hepatitis B virus (HBV) was screened to select anti-inflammatory peptides, and these novel peptides were named “Up7del1 (SEQ ID NO: 1, GGWSSK)”, “Up7del2 (SEQ ID NO: 2, GWSSK)”, “Up7del3 (SEQ ID NO: 3, WSSK)”, “Up5 (SEQ ID NO: 4, MGGWS)”, “Up6 (SEQ ID NO: 5, MGGWSS)”, and “Up7 (SEQ ID NO: 6, MGGWSSK)”.

[0161]

[0162] Figure 1 is a schematic diagram showing a hepatitis B virus-derived peptide (Up7del1).

[0163]

[0164] Reference Example 1. Assessment of disease activity and disease level in a DSS-induced colitis model.

[0165] Dextran sodium sulfate (DSS) is a water-soluble polymer compound that can induce intestinal inflammation when taken orally. DSS is highly reproducible and variable-controllable, and can induce a variety of models, including acute, chronic, and recurrent, depending on the dose and duration of administration. Therefore, it is a commonly used compound for disease model generation in inflammatory bowel disease research.

[0166]

[0167] DSS is dissolved in drinking water at a concentration of 2-5% and provided. During the period of provision, disease activity can be evaluated by observing weight loss, bloody stool, and diarrhea in mice. The criteria are as shown in Table 2 below. Table 2 shows the criteria for evaluating disease activity in the DSS-induced colitis model.

[0168] Score Weight loss (%) Diarrhea Bloody stool 0<1 None None 11-5--25-10 Mild Mild 310-20--4>20 Severe Severe

[0169]

[0170] Additionally, colonic lesions caused by DSS can be confirmed through a microscope after H&E staining, and the level of colonic inflammation can be evaluated by observing the degree of damage to the colonic crypt and the degree of immune cell infiltration.

[0171] Additionally, since the length of the colon of mice characteristically shortens when inflammation progresses due to DSS, the objective level of disease can be assessed by measuring the length of the colon after mouse autopsy.

[0172] Additionally, in immunological evaluation, the colon can be removed from the mouse, colon cells can be isolated, and the proportion and activity of immune cells can be evaluated through flow cytometry.

[0173]

[0174] Figure 2 shows a schematic diagram of the evaluation of Up7del1 efficacy using a DSS-induced colitis model.

[0175]

[0176] Reference Example 2. Induction of acute and chronic colitis models using DSS

[0177] The mouse colitis model induced by DSS can induce acute and chronic colitis depending on the administration period and dose of DSS. In the present invention, the efficacy of Up7del1 was evaluated in acute and chronic colitis. In the case of acute colitis, it was induced by dissolving 5% DSS in drinking water and providing it for 7 to 10 days. In the case of chronic colitis, it was induced by dissolving 2.5% DSS in drinking water and providing it for 7 days, then switching to normal drinking water and providing it for 7 days, repeating this process three times.

[0178]

[0179] Figure 3 shows the method of inducing acute colitis through DSS.

[0180] Figure 4 shows the method of inducing chronic colitis through DSS.

[0181]

[0182] The anti-inflammatory efficacy of Up7del1 is explained in detail through the following experimental examples.

[0183]

[0184] Experimental Example 1. Evaluation of the anti-inflammatory efficacy of Up7del1

[0185] 1.1. Evaluation of the anti-inflammatory efficacy of novel peptides in an in vitro inflammation-induced model - 1

[0186] NIH3T3 cells (W4P-LHB-NIH3T3) expressing the mutant large surface antigen of HBV, known to increase the expression of inflammatory cytokines, and the macrophage cell line J774A.1 were cultured in RPMI-1640 (10% FBS, 1% PS) for 24 h. The culture medium of W4P-LHB-NIH3T3 was added to J774A.1, and the culture was further cultured for 24 h. The supernatant was separated and subjected to ELISA for IL-6 and TNF-α.

[0187] For the group treated with the novel peptide (Up7del1), the novel peptide (Up7del1) was treated before and after adding the culture medium of W4P-LHB-NIH3T3 to J774A.1.

[0188]

[0189] Figure 5 is a graph evaluating the anti-inflammatory efficacy of Up7del1 in an inflammation-induced model using W4P-LHB-NIH3T3.

[0190]

[0191] As shown in Fig. 5, it was confirmed that when Up7del1 was treated, the inflammatory cytokines IL-6 and TNF-α decreased in proportion to the concentration compared to the untreated group (Sup).

[0192]

[0193] 1.2. Evaluation of the anti-inflammatory efficacy of novel peptides in an in vitro inflammation-induced model - 2

[0194] In the same manner as in Experimental Example 1.1, before and after adding the culture medium of W4P-LHB-NIH3T3 to J774A.1, each of the novel peptides “Up7del1, Up7del2, Up7del3, Up5, Up6, and Up7” was treated at a concentration of 10 μM, and ELISA for IL-6 was performed.

[0195]

[0196] Figure 6 is a graph evaluating the anti-inflammatory efficacy of novel peptides “Up7del1, Up7del2, Up7del3, Up5, Up6, and Up7” in an inflammation-induced model using W4P-LHB-NIH3T3.

[0197]

[0198] As shown in Fig. 6, when treated with the novel peptides “Up7del1, Up7del2, Up7del3, Up5, Up6, and Up7” according to the present invention, it was confirmed that the inflammatory cytokine IL-6 was reduced compared to the untreated group (-) and the Up11 (SEQ ID NO: 7, MGGWSSKPRQG) treated group.

[0199] The inventors of the present application confirmed that a novel peptide discovered by screening a portion of the sequence surrounding the N-terminus of PreS1 as a peptide library has excellent anti-inflammatory efficacy even when a portion of the N-terminus or C-terminus of the Up7del1 sequence is added or removed, and among them, Up7del1 has the best anti-inflammatory efficacy.

[0200]

[0201] 1.3. Evaluation of the anti-inflammatory efficacy of Up7del1 in an in vivo sepsis model

[0202] Seven-week-old female Balb / c mice were administered subcutaneously with 5 mg / kg or 0.5 mg / kg of up7del1 three times at two-day intervals, followed by intraperitoneal administration of 10 mg / kg of LPS. Blood samples were collected from the mice 2 hours after LPS administration, and serum was separated by centrifugation and subjected to ELISA for TNF-α.

[0203]

[0204] Figure 7 is a graph evaluating the anti-inflammatory efficacy of Up7del1 in an in vivo sepsis model.

[0205]

[0206] As shown in Figure 7, in mice administered Up7del1, the blood level of TNF-α, an inflammatory cytokine, was reduced, and the degree of this reduction was greater at a low dose of 0.5 mg / kg.

[0207]

[0208] Experimental Example 2. Safety Evaluation of Up7del1

[0209] 2.1. Cytotoxicity evaluation of Up7del1

[0210] Cytotoxicity evaluation by Up7del1 was performed using Neutral red uptake (NRU) assay. Mouse-derived macrophage cell line J774A.1 was seeded in 96-well at a density of 1x10 per well. 5were seeded and cultured overnight in RPMI-1640 (10% FBS, 1% Penicillin-Streptomycin) medium to attach to the plate. Up7del1 was treated to J774A.1 at various concentrations, and as a control, only the solvent without Up7del1 was treated and cultured at 37℃ for 24 and 48 hours. After culture, the cells were washed with PBS, and RPMI-1640 medium containing 40 μg / ml Neutral red was added and stained by incubating at 37℃ for 2 hours. After staining, the cells were washed with PBS and destained by treating with Destaining buffer (1% acetic acid, 49% distilled water, 50% ethanol), and then the absorbance (wavelength 540 nm) was measured using a Tecan F200 microplate reader. The cytotoxicity by Up7del1 was evaluated by normalizing the absorbance values ​​measured at the treated concentration by dividing them by the absorbance values ​​measured in the control group treated with the same amount of solvent.

[0211]

[0212] Figure 8 is a graph evaluating the cytotoxicity of Up7del1 using the Neutral red uptake (NRU) assay.

[0213]

[0214] As shown in Fig. 8, cells treated with Up7del1 did not show significant toxicity even when treated at a concentration of up to 100 μM, and this tendency was maintained even when treated for up to 48 hours.

[0215]

[0216] 2.2. Biotoxicity Evaluation of Up7del1

[0217] The biotoxicity evaluation by Up7del1 was confirmed through subcutaneous administration to 7-week-old male and female ICR mice, and a DRF toxicity test was performed upon single subcutaneous administration and a DRF toxicity test upon repeated subcutaneous administration.

[0218] Specifically, Up7del1 was administered subcutaneously at doses of 5, 10, and 20 mg / kg. For a single-dose toxicity test, it was administered once, and for a repeated-dose toxicity test, it was administered nine times (twice per week) over 4 weeks. During the administration period, the mortality rate, clinical symptoms, and body weight of the mice were evaluated, and after the final administration, an autopsy was performed to evaluate the presence of any abnormalities in the internal organs. In the case of multiple administrations, blood and urine tests were performed to check for any abnormalities.

[0219]

[0220] Figure 9 is a summary table of the results of a biotoxicity test upon single subcutaneous administration of Up7del1.

[0221]

[0222] As shown in Fig. 9, in the case of the single-dose toxicity test of Up7del1, not only was no death observed due to Up7del1 administration, but no clinical symptoms or abnormal behaviors were observed. No decrease or increase in body weight was observed, and when the organs (adrenal gland, aorta, sternal bone marrow, brain, coagulation gland, epididymis, esophagus, eye, gallbladder, hypodermic gland, heart, cecum, duodenum, ileum, jejunum, rectum, kidney, liver, lung, mesenteric lymph node, mandibular lymph node, mammary gland, skeletal muscle, optic nerve, peripheral nerve, pancreas, pituitary gland, prostate, salivary gland, seminal vesicle, skin, spinal cord, spleen, stomach, testis, thymus, thyroid / parathyroid gland, tongue, trachea, bladder, femoral tibial joint, ovary, vagina, uterus) were observed after autopsy, there were no external changes or gangrene visible to the naked eye.

[0223] Therefore, as no abnormalities were observed even at the maximum dose of 20 mg / kg used in the above test, it can be evaluated that the dose-limiting toxicity of Up7del1 upon single subcutaneous administration exceeds 20 mg / kg.

[0224]

[0225] Figure 10 is a summary table of the results of a biotoxicity test upon repeated subcutaneous administration of Up7del1.

[0226]

[0227] As shown in Fig. 10, in the Up7del1 repeat-dose toxicity test, not only was no death observed due to Up7del1 administration, but no clinical symptoms or abnormal behaviors were observed. No decrease or increase in body weight was observed, and when the organs (adrenal gland, aorta, sternal bone marrow, brain, coagulation gland, epididymis, esophagus, eyeball, gallbladder, hypodermic gland, heart, cecum, duodenum, ileum, jejunum, rectum, kidney, liver, lung, mesenteric lymph node, mandibular lymph node, mammary gland, skeletal muscle, optic nerve, peripheral nerve, pancreas, pituitary gland, prostate, salivary gland, seminal vesicle, skin, spinal cord, spleen, stomach, testis, thymus, thyroid / parathyroid gland, tongue, trachea, bladder, femoral tibial joint, ovary, vagina, uterus) were observed after autopsy, there were no external changes or gangrene visible to the naked eye. No abnormal findings were observed in blood or urine tests.

[0228] Therefore, as no abnormalities were observed even at the maximum dose of 20 mg / kg used in the above test, it can be evaluated that the dose-limiting toxicity of Up7del1 upon repeated subcutaneous administration exceeds 20 mg / kg.

[0229]

[0230] Experimental Example 3. Evaluation of Up7del1 efficacy in a DSS-induced acute colitis model.

[0231] 3.1. Evaluation of Up7del1 efficacy in an acute colitis model

[0232] Seven-week-old Balb / c female mice were subcutaneously inoculated with Up7del1 three times at two-day intervals. 5% DSS dissolved in drinking water was administered for 10 days starting on the last day of inoculation, and the mice's body weight and disease activity were assessed. Ten days after DSS administration, the mice were necropsied, the colons were isolated, and their lengths were measured. A portion of the tissue was stained with H&E for histological evaluation. The mice's body weight and disease activity were regularly monitored from the start of DSS administration until the end of the experiment.

[0233]

[0234] Figure 11 is a photograph and graph evaluating the level of relief of DSS-induced acute colitis by Up7del1. In Figure 10, item A represents weight change, item B represents change in disease activity, item C represents comparison of colon length, and item D represents histological evaluation within the colon.

[0235]

[0236] As shown in Figure 11, in mice administered Up7del1, DSS-induced weight loss was alleviated compared to the non-administered group (DSS), and similar results were observed in disease activity. In addition, the degree of inflammation-induced decrease in intestinal length was relatively small in the Up7del1-administered group. When the crypt structure within the colon was histologically evaluated using H&E staining, the crypt structure was relatively intact in the Up7del1-administered group compared to the non-administered group (DSS), in which most of the crypt structure was destroyed.

[0237]

[0238] 3.2. Evaluation of survival rate by Up7del1 in an acute colitis model

[0239] Seven-week-old Balb / c female mice were subcutaneously inoculated with 1 μg of Up7del1 three times at two-day intervals. 5% DSS dissolved in drinking water was administered starting on the last day of inoculation until a certain number of mice died. The survival of the mice was checked daily, the date of death was recorded, and a survival rate graph was created.

[0240]

[0241] Figure 12 is a graph evaluating the survival rate by Up7del1 in a DSS-induced acute colitis model.

[0242]

[0243] As shown in Figure 12, in terms of survival rate, deaths began to occur from the 12th day of DSS administration, and the same survival rate was observed in the Up7del1 administration group and the non-administration group until the 13th day, but no deaths occurred in the Up7del1 administration group after the 13th day, and on the 16th day, when all non-administration groups had died, Up7del1 showed a survival rate of over 50%.

[0244]

[0245] 3.3. Evaluation of Up7del1 efficacy according to administration dose in an acute colitis model

[0246] Seven-week-old Balb / c female mice were subcutaneously inoculated three times with 250 ng, 500 ng, and 1 μg of Up7del1 per mouse at two-day intervals. 5% DSS dissolved in drinking water was administered for 10 days starting from the last inoculation, and the body weights and disease activity of the mice were assessed. Ten days after DSS administration, the mice were necropsied, the large intestines were isolated, and the lengths of the large intestines were measured.

[0247]

[0248] Figure 13 is a graph confirming the optimal Up7del1 dosage in a DSS-induced acute colitis model. In Figure 13, item A represents colon length comparison, item B represents weight change, and item C represents change in disease activity.

[0249]

[0250] As shown in Fig. 13, when Up7del1 was administered to mice at different doses, the efficacy increased in proportion to the administered dose of Up7del1 when colon length, body weight, and disease activity were evaluated, and the highest efficacy was shown at 1 μg (0.05 mg / kg).

[0251]

[0252] Experimental Example 4. Evaluation of Up7del1 efficacy in a DSS-induced chronic colitis model.

[0253] Seven-week-old Balb / c female mice were subcutaneously inoculated with 1 μg of Up7del1 three times at two-day intervals, and 2.5% DSS dissolved in drinking water was administered for 7 days starting from the last day of inoculation. After the DSS administration period, the drinking water was replaced with normal drinking water and a 7-day rest period was provided, and before the next DSS administration, 1 μg of Up7del1 was subcutaneously inoculated three times at two-day intervals. This process was repeated three times, and the mice were necropsied, the large intestines were isolated, and the length of the large intestines was measured. The weight and disease activity of the mice were regularly monitored from the start of DSS administration to the end of the experiment.

[0254]

[0255] Figure 14 is a photograph and graph evaluating the efficacy of Up7del1 in a DSS-induced chronic colitis model. In Figure 14, item A represents colon length comparison, item B represents weight change, and item C represents change in disease activity.

[0256]

[0257] As shown in Figure 14, mice administered Up7del1 showed a reduction in DSS-induced weight loss compared to the non-administered group (DSS), and similar results were observed in disease activity. Furthermore, the degree of inflammation-induced decrease in intestinal length was relatively small in the Up7del1-administered group.

[0258]

[0259] Experimental Example 5. Identification of the Mechanism of Action of Up7del1

[0260] 5.1. Elucidating the mechanism through immunological techniques

[0261] Seven-week-old Balb / c female mice were subcutaneously inoculated with Up7del1 three times at two-day intervals, and 5% DSS dissolved in drinking water was provided for 10 days from the last inoculation. After that, the mice were necropsied, and blood, large intestine, and mesenteric lymph nodes were isolated from the mice.

[0262] The isolated colon tissue was placed in HBSS buffer containing 5 mM EDTA and 1 mM DTT and reacted at 37℃ for 30 minutes to remove the epidermal layer, and then placed in PBS containing 1 mg / ml Collagenase D and 0.5 mg / ml DNase I and reacted at 37℃ for 40 minutes to soften the tissue. The softened tissue was placed on a 70 μm nylon strainer and physically homogenized in RPMI-1640 to isolate colon cells. The isolated colon cells were reacted in RBC lysing buffer for 3 minutes, then homogenized again in RPMI-1640, and the number of cells was counted under a microscope.

[0263] The isolated lymph nodes were placed in PBS containing 0.5 mg / ml Collagenase D and 0.1 mg / ml DNase I and reacted at 37°C for 30 minutes to soften the tissue. The softened tissue was placed on a 70 μm nylon strainer and physically homogenized in RPMI-1640 to isolate immune cells.

[0264] Blood was centrifuged to separate only serum, and then ELISA for inflammatory cytokines IL-6 and TNF-α was performed.

[0265] Cells isolated from the colon and lymph nodes were seeded at 5x10 per well in a 96-well cell culture plate. 5 After seeding and culturing at 37℃ for 48 hours, the culture solution was collected and ELISA was performed.

[0266] Flow cytometry analysis was performed on cultured colonocytes and lymph node immune cells. After blocking with CD16 / 32 antibody for 10 minutes, surface antigens were stained for 30 minutes, followed by fixation and permabilization in Cytofix / Cytoperm solution (BD #554714) for 20 minutes. Intracellular antigens were then stained for 40 minutes and analyzed using a flow cytometer (BD LSRFortessa).

[0267] Some of the isolated colon cells were sorted using a flow cytometer to isolate macrophages by co-expressing F4 / 80 and CD45. These were purified using TRIzol TM RNA was extracted using (Invitrogen #15596018) and then SensiFAST TM cDNA was synthesized using a cDNA synthesis kit (#BIO-65054). The synthesized cDNA was analyzed by SensiFAST TM SYBR ® qPCR was performed using the Lo-ROX kit (#BIO-94020), and the sequences of the primers used are shown in Table 3 below.

[0268] TNF-αCXCL9ForwardGCA TGA TCC GAG ATG TGG AAC TGG (SEQ ID NO: 8)ForwardGTG GAG TTC GAG GAA CCC TAG (SEQ ID NO: 16)ReverseCGC CAC GAG CAG GAA TGA GAA G (SEQ ID NO: 9)ReverseATT GGG GCT TGG GGC AAA C (SEQ ID NO: 17)IL-6CXCL10ForwardGTT CTC TGG GAA ATC GTG GA (SEQ ID NO: 10)ForwardGTG GGA CTC AAG GGA TCC CTC (SEQ ID NO: 18)ReverseGCA TTG GAA ATT GGG GTA GG (SEQ ID NO: 11)ReverseCAG GAT AGG CTC GCA GGG ATG (SEQ ID NO: 19)IL-10CXCL11ForwardCTC GTT TGT ACC TCT CTC CG (SEQ ID NO: 12)ForwardGCT CAA GGC TTC CTT ATG TTC AAA C (SEQ ID NO: 20)ReverseATC TCC CTG GTT TCT CTT CC (SEQ ID NO: 13)ReverseCTT TGT CGC AGC CGT TAC TCG (SEQ ID NO: 21)GAPDHForwardTGA TGA CAT CAA GAA GGT GGT GAA G (SEQ ID NO: 14)ReverseTCC TTG GAG GCC ATG TAG GCC AT (SEQ ID NO: 15)

[0269]

[0270] Some colon tissues were fixed, stained with antibodies, and examined using a confocal microscope.

[0271]

[0272] Figure 15 shows changes in blood inflammatory cytokines caused by Up7del1 in a DSS-induced acute colitis model.

[0273] Figure 16 shows changes in the expression of inflammatory cytokines in colon cells by Up7del1 in a DSS-induced acute colitis model.

[0274] Figure 17 shows the change in the proportion of Th17 and Treg in the colon by Up7del1 in the DSS-induced acute colitis model.

[0275] Figure 18 shows the change in the proportion of Th17 and Treg in the mesenteric lymph nodes by Up7del1 in the DSS-induced acute colitis model.

[0276]

[0277] As shown in Figure 15, when the inflammatory cytokines IL-6 and TNF-α in the blood of mice were measured by ELISA, the group administered Up7del1 showed a significant decrease compared to the non-administered group (DSS). In addition, as shown in Figure 16, similar results were confirmed in ELISA performed with colon cell culture fluid isolated from mice. On the other hand, the anti-inflammatory cytokine IL-10 present in the colon cell culture fluid was confirmed to be increased in the Up7del1 administered group compared to the non-administered group.

[0278] In addition, as shown in Fig. 17, when the ratio of immune cells present in colon cells isolated from mice was confirmed through flow cytometry, Th17 cells known to aggravate inflammation in inflammatory bowel disease decreased, whereas Treg cells known to contribute to the alleviation of inflammation increased in the Up7del1 administration group. It was also confirmed that the ratio of Th17 to Treg significantly decreased in the Up7del1 administration group compared to the non-administered group. In addition, as shown in Fig. 18, similar results were confirmed in immune cells isolated from the mesenteric lymph nodes of mice.

[0279]

[0280] Figure 19 shows changes in the differentiation pattern of colonic macrophages by Up7del1 in a DSS-induced acute colitis model.

[0281] Figure 20 shows changes in the differentiation pattern of macrophages in lymph nodes by Up7del1 in a DSS-induced acute colitis model.

[0282] Figure 21 shows changes in the expression pattern of colonic macrophages by Up7del1 in a DSS-induced acute colitis model.

[0283]

[0284] As shown in Figure 19, when flow cytometry was performed to confirm the macrophage differentiation pattern in immune cells isolated from mice, in the case of the colon, the proportion of M2 macrophages (CD206+, CD163+), known to contribute to suppression of excessive immunity, was significantly increased in the Up7del1 treatment group compared to the control group (DSS), and the proportion of M1 macrophages (iNOS+), known to induce inflammatory responses, was confirmed to be decreased.

[0285] Additionally, as shown in Figure 20, it was confirmed that the proportion of M2 macrophages increased in the Up7del1 treatment group in the mesenteric lymph nodes.

[0286] In addition, as shown in Fig. 21, when macrophages were isolated from mouse colon cells using a flow cytometer and qPCR was performed to compare the expression patterns, the expression of chemokines CXCL9, CXCL10, CXCL11 and anti-inflammatory cytokine IL-10 increased in the Up7del1 treatment group, and among these, the expression of CXCL9 increased most significantly. On the other hand, it was confirmed that the inflammatory cytokines IL-6 and TNF-α decreased.

[0287]

[0288] As described above, we confirmed that the number of regulatory T cells increased in the colon of mice administered Up7del1, and to elucidate the mechanism, we compared the colons of mice stained with antibodies using a confocal microscope.

[0289]

[0290] Figure 22 shows an increase in CXCL9+ macrophages and CXCR3+ regulatory T cells in the colon following Up7del1 treatment as confirmed by confocal microscopy.

[0291] Figure 23 shows an increase in CXCL9+ macrophages and CXCR3+ regulatory T cells in the colon following Up7del1 treatment as confirmed by flow cytometry.

[0292]

[0293] As a result, as shown in Figure 22, it was confirmed that the expression of macrophages expressing CXCL9 increased in the large intestine of mice, and at the same time, the number of regulatory T cells expressing CXCR3 increased.

[0294] In addition, as shown in Figure 23, flow cytometry analysis confirmed that macrophages expressing CXCL9 and regulatory T cells expressing CXCR3 increased in the group treated with Up7del1 among the immune cells in the colon.

[0295] Considering that CXCL9 is a chemokine that binds to CXCR3 and is involved in the influx of immune cells, these results suggest that the increased expression of CXCL9 in macrophages by Up7del1 induces the influx of regulatory T cells expressing CXCR3 into the colon, thereby alleviating and resolving the inflammatory response.

[0296] In particular, since regulatory T cells are known to be important immune cells in alleviating inflammation in inflammatory diseases, Up7del1 can be useful in the prevention and treatment of various inflammatory diseases such as inflammatory bowel disease, systemic lupus erythematosus, rheumatoid arthritis, and atopic dermatitis.

[0297]

[0298] 5.2. Elucidating the mechanism through CXCL9 blocking

[0299] Seven-week-old Balb / c female mice were intraperitoneally administered 200 μg of monoclonal CXCL9 neutralizing antibody (BioXcell, #BE0309) or its control polyclonal IgG (BioXcell, #BE0091) at two-day intervals for a total of six doses. Up7del1 was administered subcutaneously at 1 μg at two-day intervals for a total of three doses, and 5% DSS dissolved in drinking water was administered for 10 days starting from the last day of inoculation. After necropsy, blood, colon, and mesenteric lymph nodes were isolated from the mice. The weight and disease activity of the mice were regularly monitored during the DSS administration period.

[0300]

[0301] Figure 24 shows the experimental schedule and administration schedule to confirm the decrease in the therapeutic efficacy of Up7del1 on DSS-induced acute colitis when CXCL9 is blocked.

[0302]

[0303] The separated colon was measured in length, and some of it was stained with H&E and observed under a microscope to histologically evaluate the degree of inflammation and damage.

[0304] The isolated colon tissue was placed in HBSS buffer containing 5 mM EDTA and 1 mM DTT and reacted at 37℃ for 30 minutes to remove the epidermal layer, and then placed in PBS containing 1 mg / ml Collagenase D and 0.5 mg / ml DNase I and reacted at 37℃ for 40 minutes to soften the tissue. The softened tissue was placed on a 70 μm nylon strainer and physically homogenized in RPMI-1640 to isolate colon cells. The isolated colon cells were reacted in RBC lysing buffer for 3 minutes, then homogenized again in RPMI-1640, and the number of cells was counted under a microscope.

[0305] The isolated lymph nodes were placed in PBS containing 0.5 mg / ml Collagenase D and 0.1 mg / ml DNase I and reacted at 37°C for 30 minutes to soften the tissue. The softened tissue was placed on a 70 μm nylon strainer and physically homogenized in RPMI-1640 to isolate immune cells.

[0306] Flow cytometry analysis was performed on cultured colonocytes and lymph node immune cells. After blocking with CD16 / 32 antibody for 10 minutes, surface antigens were stained for 30 minutes, followed by fixation and permabilization in Cytofix / Cytoperm solution (BD #554714) for 20 minutes. Intracellular antigens were then stained for 40 minutes and analyzed using a flow cytometer (BD LSRFortessa).

[0307]

[0308] Figure 25 is a graph evaluating the decrease in the efficacy of Up7del1 in treating DSS-induced acute colitis by CXCL9 blocking through changes in body weight.

[0309] Figure 26 is a graph evaluating the decrease in the therapeutic efficacy of Up7del1 in DSS-induced acute colitis by CXCL9 blocking through changes in disease activity.

[0310] Figure 27 is a photograph and graph evaluating the reduction in the efficacy of Up7del1 in treating DSS-induced acute colitis by CXCL9 blocking through comparison of colon length.

[0311] Figure 28 is a graph histologically evaluating the decrease in the efficacy of Up7del1 in treating DSS-induced acute colitis when CXCL9 is blocked.

[0312]

[0313] As shown in Figure 25, when Up7del1 and polyclonal IgG were administered, the weight loss of the mice was inhibited and the disease activity was reduced compared to the negative control group (DSS). However, when CXCL9 was blocked using a CXCL9 neutralizing antibody, not only was the body weight reduced to the same extent as the negative control group despite Up7del1 administration, but as shown in Figure 26, the disease activity was also confirmed to increase to the same extent as the negative control group.

[0314] In addition, as shown in Figure 27, the length of the colon due to inflammation increased when Up7del / IgG was administered, whereas this increase was significantly reduced when CXCL9 was blocked.

[0315] In addition, as shown in Figure 28, in the histological evaluation, it was confirmed that inflammation and crypt damage in the colon tissue of the Up7del1 / IgG administration group were significantly reduced compared to the negative control group, whereas this effect disappeared when CXCL9 was blocked.

[0316] These results can be evaluated as proof that the therapeutic efficacy of Up7del1 in the DSS-induced colitis model is dependent on CXCL9.

[0317]

[0318] Figures 29 to 32 are graphs evaluating the reduction in anti-inflammatory efficacy of Up7del1 in DSS-induced acute colitis when CXCL9 is blocked.

[0319]

[0320] As shown in Figure 29, when ELISA was performed on colon cell culture fluid, it was confirmed that the inflammatory cytokines IL-6 and TNF-α, which had decreased in the Up7del1 / IgG administration group, increased again in the CXCL9 neutralizing antibody administration group, and the anti-inflammatory cytokine IL-10, which had decreased, increased again.

[0321] As shown in Figure 30, flow cytometry analysis confirmed that macrophages expressing CXCL9 in the colon and lymph nodes significantly increased in the Up7del1 / IgG administration group, but decreased again in the CXCL9 neutralizing antibody administration group.

[0322] Furthermore, as shown in Figure 31, the ratio of Th17 cells and Treg cells, known to exacerbate inflammation in inflammatory bowel disease, was confirmed in colonocytes, and was significantly reduced in the Up7del1 / IgG administration group, but increased again in the CXCL9 neutralizing antibody administration group. As shown in Figure 32, these results were also observed in mesenteric lymph nodes.

[0323] Considering that CXCL9 is a chemokine that binds to CXCR3 and is involved in the influx of immune cells, these results suggest that the increased expression of CXCL9 in macrophages by Up7del1 induces the influx of regulatory T cells expressing CXCR3 into the colon, thereby alleviating and resolving the inflammatory response.

[0324] In particular, since regulatory T cells are known to be important immune cells in alleviating inflammation in inflammatory diseases, Up7del1 can be useful in the prevention and treatment of various inflammatory diseases such as inflammatory bowel disease, systemic lupus erythematosus, rheumatoid arthritis, and atopic dermatitis.

[0325]

[0326] From the above results, it can be seen that Up7del1 has high safety and has a high therapeutic index by showing high anti-inflammatory activity in animal models even at low doses. In addition, considering the above-mentioned strong anti-inflammatory effect and mechanism of action of Up7del1 (inducing the expression of CXCL9 in macrophages and attracting CXCR3+ T cells), it can be seen that Up7del1 has excellent preventive and therapeutic effects on inflammatory diseases.

[0327]

[0328] From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without altering its technical concept or essential characteristics. In this regard, it should be understood that the experimental examples and embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be interpreted as encompassing all changes or modifications derived from the meaning and scope of the following claims and their equivalent concepts, rather than the detailed description above.

Claims

1. An isolated peptide consisting of an amino acid sequence having at least 65% sequence identity with the amino acid sequence of sequence number 1, or a solvate or salt thereof.

2. In the first paragraph, the peptide is a peptide consisting of a sequence of 4 to 7 amino acids, or a solvate or salt thereof.

3. In the first paragraph, the peptide Consisting of an amino acid sequence of sequence number 1; It consists of an amino acid sequence in which any one amino acid in the amino acid sequence of sequence number 1 is replaced with another amino acid; It consists of an amino acid sequence having one or two amino acid sequences additionally linked or deleted at the N-terminus of the amino acid sequence of sequence number 1 or the substituted amino acid sequence; or consisting of an amino acid sequence having one or two amino acid sequences additionally linked or deleted at the C-terminus of the amino acid sequence of sequence number 1 or the substituted amino acid sequence; or A peptide consisting of an amino acid sequence having one or two amino acid sequences additionally linked or deleted at the N-terminus of the amino acid sequence of sequence number 1 or the substituted amino acid sequence, and one or two amino acid sequences additionally linked or deleted at the C-terminus, or a solvate or salt thereof.

4. In the first paragraph, the peptide Consisting of an amino acid sequence of sequence number 1; It consists of an amino acid sequence in which any one amino acid in the amino acid sequence of sequence number 1 is replaced with another amino acid; It consists of an amino acid sequence having one or two amino acid sequences deleted at the N-terminus of the amino acid sequence of sequence number 1 or the substituted amino acid sequence; or consisting of an amino acid sequence having methionine additionally bonded to the N-terminus of the amino acid sequence of sequence number 1 or the substituted amino acid sequence; or A peptide consisting of an amino acid sequence having an amino acid sequence of sequence number 1 or a substituted amino acid sequence in which a methionine is additionally bonded to the N-terminus and one or two amino acid sequences are deleted from the C-terminus, or a solvate or salt thereof.

5. In the first paragraph, the peptide is a peptide derived from hepatitis B virus (HBV) PreS1, or a solvate or salt thereof.

6. In the first paragraph, the peptide is a peptide having anti-inflammatory activity, or a solvate or salt thereof.

7. A peptide, or a solvate or salt thereof, in claim 1, wherein the N-terminus of the peptide is bound to any one protecting group selected from the group consisting of an acetyl group, a fluoreonylmethoxycarbonyl group, a formyl group, a palmitoyl group, a myristyl group, a stearyl group, a butoxycarbonyl group, an aryloxycarbonyl group, and polyethylene glycol (PEG).

8. A peptide, or a solvate or salt thereof, in the first paragraph, wherein the C-terminus of the peptide is bound to any one protecting group selected from the group consisting of an amino group (-NH2), a tertiary alkyl group, and an azide (-NHNH2).

9. A pharmaceutical composition for preventing or treating inflammatory diseases, comprising the peptide of paragraph 1, or a solvate or salt thereof, as an active ingredient.

10. In paragraph 9, the inflammatory disease is inflammatory bowel disease, irritable bowel syndrome, sepsis, septic shock, chronic inflammatory disease caused by chronic viral or bacterial infection, pancreatitis, gastric ulcer, gastritis, rhinitis, bronchitis, periodontitis, inflammatory skin disease, atopic dermatitis, encephalitis, chronic obstructive pulmonary disease, pulmonary fibrosis, undifferentiated spondyloarthropathy, undifferentiated arthropathy, arthritis, inflammatory osteolysis, inflammatory collagen vascular disease, nephritis, glomerulonephritis, type 1 diabetes, rheumatoid arthritis, reactive arthritis, osteoarthritis, psoriasis, scleroderma, osteoporosis, atherosclerosis, myocarditis, endocarditis, pericarditis, cystic fibrosis, Hashimoto's thyroiditis, Graves' disease, leprosy, syphilis, Lyme disease, borreliosis, neuro-borreliosis, tuberculosis, sarcoidosis, lupus, A pharmaceutical composition comprising at least one selected from the group consisting of discoid lupus, chilblain lupus, lupus nephritis, systemic lupus erythematosus, macular degeneration, uveitis, Crohn's disease, Sjogren's syndrome, fibromyalgia, chronic fatigue syndrome, chronic fatigue immunodeficiency syndrome, myalgic encephalomyelitis, amyotrophic lateral sclerosis, Parkinson's disease, and multiple sclerosis.

11. A pharmaceutical composition according to claim 10, wherein the inflammatory bowel disease is at least one selected from the group consisting of Crohn's disease, ulcerative colitis, collagenous colitis, lymphocytic colitis, ischemic colitis, diversionary colitis, colitis, hemorrhagic rectal ulcer, Behcet's disease, inflammatory enteritis, chronic enteritis, acute enteritis, intestinal tuberculosis, ileal pouchitis, bacterial enteritis, viral enteritis, amoebic enteritis, and ischemic colitis.

12. A pharmaceutical composition according to claim 9, wherein the peptide has anti-inflammatory activity by inducing the expression of CXCL9 in macrophages and attracting CXCR3+ T cells.

13. A pharmaceutical composition according to claim 9, wherein the N-terminus of the peptide is bound to any one protecting group selected from the group consisting of an acetyl group, a fluoreonylmethoxycarbonyl group, a formyl group, a palmitoyl group, a myristyl group, a stearyl group, a butoxycarbonyl group, an aryloxycarbonyl group, and polyethylene glycol (PEG).

14. A pharmaceutical composition according to claim 9, wherein the C-terminus of the peptide is bound to any one protecting group selected from the group consisting of an amino group (-NH2), a tertiary alkyl group, and an azide (-NHNH2).

15. A health functional food for preventing or improving inflammatory diseases, comprising the peptide of paragraph 1, or a solvate or salt thereof.

16. A feed composition for preventing or improving inflammatory diseases, comprising the peptide of paragraph 1 or 2, or a solvate or salt thereof.

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