Peptide compositions capable of binding to lanthionine synthase C-like protein (LanCL) and uses thereof

Abstract

The present invention provides various peptide compositions capable of binding to Lanthionine Synthase C-like protein (LanCL) and having analgesic, anti-inflammatory and antimicrobial properties. The compositions comprise a peptide of formula (I): X1-X2-X3-X4-X5-X6, where X represents a specific group of amino acids; the peptide is 3-20 amino acids in length; does not contain the sequence CRSRPVESC, CRSVEGSCG or CRIIHNNNC; and is not a linear peptide containing the sequence EQLERALNSS.

Description

[Technical field]

[0001] The present invention relates generally to peptides suitable for the treatment of conditions such as pain, inflammatory conditions and respiratory infections and their uses. [Background technology]

[0002] All references, including patents or patent applications, cited in this specification are incorporated herein by reference to enable a complete understanding of the invention. However, such references should not be read as an acknowledgement that any of them form part of the general knowledge in the art in Australia or anywhere else.

[0003] As described by Lee et al. (Int J Mol Sci., 2019, 20(10):2383), protein-protein interactions (PPIs) are the basis of almost all cellular processes. Those biochemical processes are often composed of activated receptors that indirectly or directly control a series of cell signaling events that regulate the transcription of nucleic acids and / or post-translational modifications of translated proteins. Drugs that specifically bind to such receptors can act as agonists or antagonists, leading to downstream consequences on cellular behavior. Thus, peptides and small molecules that inhibit PPIs are sought as therapeutic agents, as they may regulate disease-related protein interactions. The authors point out that better identification of targetable disease-related PPIs and optimization of peptide-drug binding properties will be key to clinical success. However, understanding the molecular recognition mechanisms of PPIs and revealing their binding affinities are complex challenges for computational biologists and protein biochemists alike, mainly due to the superiority of small molecules in binding to deep folding pockets of proteins compared to the large, flat, hydrophobic binding interfaces that are commonly present at PPI complex interfaces. Although antibodies are generally more effective at recognizing PPI interfaces, they generally cannot penetrate cell membranes to reach and recognize intracellular targets. The authors point out that in recent years, peptides that are up to five times larger than small molecule drugs and have balanced conformational flexibility and binding affinity have attracted much attention. For example, cyclic peptides have the properties of small molecule drugs with long-term in vivo stability while maintaining the tight binding affinity and minimal toxicity of antibodies.

[0004] de la Torre and Albericio (2020; Molecules; 25(10); p. 2293) report that the field of peptide-based drug discovery has recently experienced a remarkable boom, noting that from 2015 to 2019, the US Food and Drug Administration (FDA) approved 208 new drugs, of which 150 were new chemical entities, 58 were biologics, and 15 were peptides or peptide-containing molecules. Among these are ixazomib (an N-acylated C-boronic acid dipeptide for the treatment of multiple myeloma), adlixin (a 34 amino acid analogue of parathyroid hormone-related protein for the treatment of osteoporosis), etelcalcetide (Ac-DCys-DAla-(DArg)3-DAla-DArg-NH2 linked to L-Cys via a disulfide bridge for the treatment of hyperparathyroidism) and afamelanotide (a 13 amino acid linear peptide analogue of α-melanocyte stimulating hormone (αMSH) for the treatment of skin damage and pain). The authors point out that while oncology, metabolism and endocrinology are the most common indications for peptide-based therapeutics approved by the FDA, cardiovascular, gastroenterology, bone disease, dermatology and sexual dysfunction are also indications targeted by FDA-approved peptide-based therapeutics.

[0005] Compared with small molecules such as proteins and antibodies, peptides are a unique class of pharmaceutical compounds based on their unique biochemical and therapeutic properties. In addition to peptide-based natural hormone analogues, peptides have been developed as drug candidates to disrupt protein-protein interactions (PPIs) and target or inhibit intracellular molecules such as receptor tyrosine kinases. Such strategies have made peptide therapeutics a leading industry with nearly 20 new peptide-based clinical trials per year. In fact, there are currently more than 400 peptide drugs in clinical development worldwide, and more than 60 have already been approved for clinical use in the United States, Europe, and Japan.

[0006] Although peptide-based therapeutics have made considerable progress, most of peptide-based therapeutics are limited to treating certain diseases and conditions that correspond to the PPIs and cell signaling pathways that these peptide-based therapeutics target.Therefore, there is a continuing need for broad-spectrum peptide-based therapeutic strategies that can effectively alleviate multiple diseases, conditions or their symptoms, including those related to cellular aging, injury or stress.The present invention lifts or at least partially alleviates this limitation by providing therapeutic peptides with broad-spectrum activity, such as analgesic activity, anti-inflammatory activity and antimicrobial activity. Summary of the Invention

[0007] In an embodiment disclosed herein, a peptide capable of binding to a Lanthionine Synthase C-like (LanCL) protein, comprising: The peptide has an amino acid sequence of formula (I): X1-X2-X3-X4-X5-X6(I) [In the formula, X1 is selected from the group consisting of lysine, arginine and histidine, or X1 is absent; X2 is selected from the group consisting of alanine, valine, leucine, isoleucine, proline, phenylalanine, cysteine, tyrosine and serine; X3 is selected from the group consisting of glycine, alanine, valine, leucine and isoleucine; X4 is selected from the group consisting of serine, cysteine, threonine, asparagine, arginine, glutamine, tyrosine, aspartic acid, lysine, glutamic acid, proline and histidine, or X4 is absent; X5 is selected from the group consisting of serine, cysteine, threonine, asparagine, arginine, glutamine, tyrosine, lysine, histidine and glycine, or X5 is absent; and X6 is selected from the group consisting of serine, cysteine, threonine, asparagine, glutamine, tyrosine and histidine, or X6 is absent. Including, the peptide is 3 to 20 amino acids in length, The amino acid sequence of the peptide does not include CRSRPVESC, CRSVEGSCG, or CRIIHNNNC; and The peptide is provided, wherein the peptide is not a linear peptide comprising the amino acid sequence EQLERALNSS.

[0008] In another embodiment disclosed herein, a peptide capable of binding to a Lanthionine Synthase C-like (LanCL) protein, comprising: The peptide has an amino acid sequence of formula (I): X1-X2-X3-X4-X5-X6(I) [In the formula, X1 is selected from the group consisting of lysine, arginine and histidine; X2 is selected from the group consisting of alanine, valine, leucine, isoleucine, proline, phenylalanine, cysteine, tyrosine and serine; X3 is selected from the group consisting of glycine, alanine, valine, leucine and isoleucine; X4 is selected from the group consisting of serine, cysteine, threonine, asparagine, arginine, glutamine, tyrosine, aspartic acid, lysine, glutamic acid, proline and histidine, or X4 is absent; X5 is selected from the group consisting of serine, cysteine, threonine, asparagine, arginine, glutamine, tyrosine, lysine, histidine and glycine, or X5 is absent; and X6 is selected from the group consisting of serine, cysteine, threonine, asparagine, glutamine, tyrosine and histidine, or X6 is absent. Including, the peptide is 3 to 20 amino acids in length, The amino acid sequence of the peptide does not include CRSRPVESC, CRSVEGSCG, or CRIIHNNNC; and The peptide is provided, wherein the peptide is not a linear peptide comprising the amino acid sequence EQLERALNSS. [Brief description of the drawings]

[0009] [Figure 1] Figure 1 shows the effect of peptide of SEQ ID NO: 1 on the viability of taxol-stressed A549 adenocarcinoma human alveolar basal epithelial cells. Cells were treated with LanCL1 siRNA (100 nM, 48 hours) to knock down LanCL1 expression. Cells were then incubated in the presence of taxol (IC50 approx. 350 μM). Incubation was performed in the presence of vehicle alone (dimethyl sulfoxide; DMSO) or in the presence of peptide of SEQ ID NO: 1 at 1, 5, 25, 50 and 100 μM concentrations (diluted in DMSO). The Y-axis shows relative light units (RLU) and the X-axis shows the concentration of peptide. [Diagram 2] Figure 2 shows the effect of peptide of SEQ ID NO: 9 on the viability of taxol-stressed A549 cells. Cells were treated with taxol (IC50 approx. 350 μM) in the presence of vehicle alone (DMSO) or in the presence of peptide of SEQ ID NO: 9 at concentrations of 1, 5, 25, 50 and 100 μM (diluted in DMSO). The Y-axis shows the relative light units (RLU) and the X-axis shows the concentration of peptide. [Diagram 3] Figure 3 shows the effect of peptides RSVEGS (SEQ ID NO: 9), SVEGS (SEQ ID NO: 62) and ALNSS (SEQ ID NO: 63) on ipsilateral paw withdrawal threshold (PWT; grams) in the rat Chung model of neuropathic pain. *P<0.05, **P<0.01, ***P<0.001 compared to the vehicle group (one-way ANOVA; n=6 per group). [Figure 4] FIG. 4 shows the effect of peptides RSVEGS (SEQ ID NO: 9), SVEGS (SEQ ID NO: 62) and ALNSS (SEQ ID NO: 63) on contralateral paw withdrawal threshold (PWT; grams) in the rat Chung model of neuropathic pain (one-way ANOVA; n=6 per group). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which this invention belongs. Although any method and material similar or equivalent to those described herein can be used to carry out or test the present invention, the preferred method and material are described. For the purpose of the present invention, the following terms are defined below.

[0011] The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or one or more than one element.

[0012] As used herein, the term "about" refers to a quantity, level, value, dimension, size or amount that varies by up to 10% (e.g., 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1%) from a reference quantity, level, value, dimension, size or amount.

[0013] Throughout this specification, unless the context otherwise requires, the terms "comprise", "comprises" and "comprising" are understood to mean the inclusion of a stated step or element or group of steps or elements but not to the exclusion of other steps or elements or groups of steps or elements.

[0014] peptide The inventors had previously identified a molecular target (lanthionine synthase C-like protein; LanCL) for a new class of cyclic peptide molecules previously recognized for analgesic and other therapeutic properties. This work is described in WO 2021 / 127752. The inventors then identified a novel consensus sequence of peptides (formula (I)) that unexpectedly retains at least some of the biological activities previously recognized for this class of cyclic LanCL-binding peptides, including analgesic, anti-inflammatory and antimicrobial activities. Furthermore, the inventors unexpectedly found that many peptides containing this consensus sequence retain biological activity regardless of whether they are present in the form of a cyclic or linear peptide. Thus, in an embodiment disclosed herein, a peptide capable of binding to a lanthionine synthase C-like (LanCL) protein, comprising: The peptide has an amino acid sequence of formula (I): X1-X2-X3-X4-X5-X6(I) [In the formula, X1 is selected from the group consisting of lysine, arginine and histidine; X2 is selected from the group consisting of alanine, valine, leucine, isoleucine, proline, phenylalanine, cysteine, tyrosine and serine; X3 is selected from the group consisting of glycine, alanine, valine, leucine and isoleucine; X4 is selected from the group consisting of serine, cysteine, threonine, asparagine, arginine, glutamine, tyrosine, aspartic acid, lysine, glutamic acid, proline and histidine, or X4 is absent; X5 is selected from the group consisting of serine, cysteine, threonine, asparagine, arginine, glutamine, tyrosine, lysine, histidine and glycine, or X5 is absent; and X6 is selected from the group consisting of serine, cysteine, threonine, asparagine, glutamine, tyrosine and histidine, or X6 is absent. Including, the peptide is 3 to 20 amino acids in length, The amino acid sequence of the peptide does not include CRSRPVESC, CRSVEGSCG, or CRIIHNNNC; and The peptide is provided, wherein the peptide is not a linear peptide comprising the amino acid sequence EQLERALNSS.

[0015] In one embodiment, X1 is arginine. In one embodiment, the peptide is not a linear peptide comprising the amino acid sequence QEQLERALNSS.

[0016] The inventors have also demonstrated that the peptides described herein unexpectedly retain at least some of the biological activity previously observed for this class of cyclic LanCL-binding peptides, including analgesic, anti-inflammatory and antimicrobial activity, even in the absence of X1. Thus, in one embodiment described herein, X1 is absent.

[0017] In another embodiment disclosed herein, a peptide capable of binding to a Lanthionine Synthase C-like (LanCL) protein, comprising: The peptide has an amino acid sequence of formula (I): X1-X2-X3-X4-X5-X6(I) [In the formula, X1 is selected from the group consisting of lysine, arginine and histidine, or X1 is absent; X2 is selected from the group consisting of alanine, valine, leucine, isoleucine, proline, phenylalanine, cysteine, tyrosine and serine; X3 is selected from the group consisting of glycine, alanine, valine, leucine and isoleucine; X4 is selected from the group consisting of serine, cysteine, threonine, asparagine, arginine, glutamine, tyrosine, aspartic acid, lysine, glutamic acid, proline and histidine, or X4 is absent; X5 is selected from the group consisting of serine, cysteine, threonine, asparagine, arginine, glutamine, tyrosine, lysine, histidine and glycine, or X5 is absent; and X6 is selected from the group consisting of serine, cysteine, threonine, asparagine, glutamine, tyrosine, and histidine, or X6 is absent. Including, the peptide is 3 to 20 amino acids in length, The amino acid sequence of the peptide does not include CRSRPVESC, CRSVEGSCG, or CRIIHNNNC; and The peptide is provided, wherein the peptide is not a linear peptide comprising the amino acid sequence EQLERALNSS.

[0018] In one embodiment, the peptide is not a linear peptide comprising the amino acid sequence QEQLERALNSS.

[0019] In one embodiment, the amino acid sequence of the peptide does not include CRSRPVESC, CRSVEGSCG, CRIIHNNNC, CRRFVESSCA, or CRIVYDSNC.

[0020] In one embodiment, X2 is selected from the group consisting of alanine, isoleucine, proline, phenylalanine and serine.

[0021] In one embodiment, X3 is selected from the group consisting of valine, leucine and isoleucine.

[0022] In one embodiment, X4 is selected from the group consisting of asparagine, glutamic acid, and histidine, or X4 is absent. In one embodiment, X4 is selected from the group consisting of asparagine, glutamic acid, proline, and histidine. In one embodiment, X4 is absent.

[0023] In one embodiment, X5 is selected from the group consisting of serine, asparagine and glycine, or X5 is absent. In one embodiment, X5 is selected from the group consisting of serine, asparagine and glycine. In one embodiment, X5 is absent.

[0024] In one embodiment, X6 is serine or asparagine, or X6 is absent. In one embodiment, X6 is serine or asparagine. In one embodiment, X6 is absent.

[0025] In one embodiment, X1 is selected from the group consisting of lysine, arginine and any conservative amino acid substitutions thereof, X2 is selected from the group consisting of alanine, isoleucine, proline, serine and any conservative amino acid substitutions thereof, X3 is selected from the group consisting of valine, leucine, isoleucine and any conservative amino acid substitutions thereof, X4 is selected from the group consisting of asparagine, glutamic acid, proline and any conservative amino acid substitutions thereof or X4 is absent, X5 is selected from the group consisting of serine, glutamine and any conservative amino acid substitutions thereof or X5 is absent, and X6 is serine or a conservative amino acid substitution thereof or X6 is absent.

[0026] In one embodiment, X1 is absent or selected from the group consisting of lysine, arginine and any conservative amino acid substitution thereof, X2 is selected from the group consisting of alanine, isoleucine, proline, serine and any conservative amino acid substitution thereof, X3 is selected from the group consisting of valine, leucine, isoleucine and any conservative amino acid substitution thereof, X4 is selected from the group consisting of asparagine, glutamic acid, proline and any conservative amino acid substitution thereof or X4 is absent, X5 is selected from the group consisting of serine, glutamine and any conservative amino acid substitution thereof or X5 is absent, and X6 is serine or a conservative amino acid substitution thereof or X6 is absent.

[0027] In one embodiment, X1 is lysine or arginine, X2 is selected from the group consisting of alanine, isoleucine, proline and serine, X3 is selected from the group consisting of valine, leucine and isoleucine, X4 is asparagine, proline or glutamic acid or X4 is absent, X5 is serine or glutamine or X5 is absent, and X6 is serine or X6 is absent.

[0028] In one embodiment, X1 is absent or X1 is lysine or arginine, X2 is selected from the group consisting of alanine, isoleucine, proline and serine, X3 is selected from the group consisting of valine, leucine and isoleucine, X4 is asparagine, proline or glutamic acid or X4 is absent, X5 is serine or glutamine or X5 is absent and X6 is serine or X6 is absent.

[0029] In one embodiment, the peptide comprises an amino acid sequence selected from the group consisting of RAL, RALN, RALNS, RALNSS, RSV, RSVE, RSVEG, RSVEGS, RPV, RPVE, RPVES, RPVESS, RII, RIIH, RIIHN and RIIHNN.

[0030] In one embodiment, the peptide consists of an amino acid sequence selected from the group consisting of RAL, RALN, RALNS, RALNSS, RSV, RSVE, RSVEG, RSVEGS, RPV, RPVE, RPVES, RPVESS, RII, RIIH, RIIHN and RIIHNN.

[0031] In one embodiment, the peptide comprises the amino acid sequence ALNSS. In one embodiment, the peptide consists of the amino acid sequence ALNSS.

[0032] In one embodiment, the peptide comprises the amino acid sequence KAPLPRS.In one embodiment, the peptide consists of the amino acid sequence KAPLPRS.

[0033] In one embodiment, the peptide comprises the amino acid sequence RALNSS.

[0034] In one embodiment, the peptide consists of the amino acid sequence RALNSS.

[0035] In one embodiment, the peptide comprises the amino acid sequence CRALNSSC.

[0036] In one embodiment, the peptide consists of the amino acid sequence CRALNSSC.

[0037] In one embodiment, the peptide is capable of competing for binding to LanCL with a peptide consisting of the amino acid sequence CRSVEGSCG.

[0038] As described elsewhere herein, the present inventors have unexpectedly shown that peptides comprising the amino acid sequence of formula (I) that are as little as three amino acids long retain biological activity. In one embodiment disclosed herein, the peptide is 3-19 amino acid residues long, preferably 3-18 amino acid residues long, preferably 3-17 amino acid residues long, preferably 3-16 amino acid residues long, preferably 3-15 amino acid residues long, preferably 3-14 amino acid residues long, preferably 3-13 amino acid residues long, preferably 3-12 amino acid residues long, preferably 3-11 amino acid residues long, preferably 3-10 amino acid residues long, preferably 3-9 amino acid residues long, preferably 3-8 amino acid residues long, preferably 3-7 amino acid residues long, preferably 3-6 amino acid residues long, preferably 3-5 amino acid residues long, preferably 3-4 amino acid residues long, or preferably 3 amino acid residues long. In one embodiment, the peptide is 20 amino acid residues long. In one embodiment, the peptide is 19 amino acid residues long. In one embodiment, the peptide is 18 amino acid residues long. In one embodiment, the peptide is 17 amino acid residues long. In one embodiment, the peptide is 16 amino acid residues long. In one embodiment, the peptide is 15 amino acid residues long. In one embodiment, the peptide is 14 amino acid residues long. In one embodiment, the peptide is 13 amino acid residues long. In one embodiment, the peptide is 12 amino acid residues long. In one embodiment, the peptide is 11 amino acid residues long. In one embodiment, the peptide is 10 amino acid residues long. In one embodiment, the peptide is 9 amino acid residues long. In one embodiment, the peptide is 8 amino acid residues long. In one embodiment, the peptide is 7 amino acid residues long. In one embodiment, the peptide is 6 amino acid residues long. In one embodiment, the peptide is 5 amino acid residues long. In one embodiment, the peptide is 4 amino acid residues long. In one embodiment, the peptide is 3 amino acid residues long.

[0039] The peptides described herein may suitably contain naturally occurring amino acid residues, either proteinogenic or non-proteinogenic. These amino acids typically have the L-stereochemistry. Naturally occurring amino acids are shown in Table 1 below.

[0040] [Table 1]

[0041] As used herein, the term "alkyl" refers to a straight-chain or branched-chain saturated hydrocarbon group having from 1 to 10 carbon atoms. Where appropriate, the alkyl group may have a specific number of carbon atoms, such as, for example, C 1-6 Alkyl includes alkyl groups having 1, 2, 3, 4, 5 or 6 carbon atoms in a straight or branched arrangement. Examples of suitable alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, 4-methylbutyl, n-hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 5-methylpentyl, 2-ethylbutyl, 3-ethylbutyl, heptyl, octyl, nonyl, and decyl.

[0042] As used herein, the term "alkenyl" refers to a straight or branched chain hydrocarbon group having one or more double bonds between carbon atoms and having from 2 to 10 carbon atoms. Where appropriate, an alkenyl group may have a specific number of carbon atoms. For example, C2-C6 in "C2-C6 alkenyl" includes groups having 2, 3, 4, 5 or 6 carbon atoms in a straight or branched chain arrangement. Examples of suitable alkenyl groups include, but are not limited to, ethenyl, propenyl, isopropenyl, butenyl, butadienyl, pentenyl, pentadienyl, hexenyl, hexadienyl, heptenyl, octenyl, nonenyl, and decenyl.

[0043] As used herein, the term "alkynyl" refers to a straight or branched chain hydrocarbon group having one or more triple bonds and having from 2 to 10 carbon atoms. Where appropriate, an alkynyl group may have a specific number of carbon atoms. For example, C2-C6 in "C2-C6 alkynyl" includes groups having 2, 3, 4, 5 or 6 carbon atoms in a straight or branched arrangement. Examples of suitable alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl and hexynyl.

[0044] As used herein, the term "cycloalkyl" refers to saturated and unsaturated (but not aromatic) cyclic hydrocarbons. The cycloalkyl ring may contain a specific number of carbon atoms. For example, a 3- to 8-membered cycloalkyl group contains 3, 4, 5, 6, 7, or 8 carbon atoms. Examples of suitable cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, and cyclooctyl.

[0045] As used herein, the term "aryl" is intended to mean a stable monocyclic, bicyclic, or tricyclic carbocyclic ring system having up to seven atoms in each ring, in which at least one ring is aromatic. Examples of such aryl groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, fluorenyl, phenanthrenyl, biphenyl, and binaphthyl.

[0046] In one embodiment, the peptide comprises one or more D-amino acids. In another embodiment, one or more of the amino acids of formula (I) are D-amino acids.

[0047] As noted elsewhere herein, the inventors have unexpectedly found that the peptides described herein retain biological activity whether present in the form of a cyclic or linear peptide. Thus, in one embodiment, the peptide is a linear peptide. In other embodiments, the peptide is a cyclic peptide. Those skilled in the art will be familiar with suitable methods for forming cyclic peptides, illustrative examples of which are described by Choi and Joo (Biomol Ther (Seoul). 2020; 28(1): 18-24), the contents of which are incorporated herein by reference.

[0048] In one embodiment, the peptide is cyclized by a disulfide bond between two cysteine ​​residues. In one embodiment, the disulfide bond is formed between two cysteine ​​residues, where the two cysteine ​​residues are located immediately adjacent to the C-terminal (X6) and N-terminal (X1) residues of formula (I). That is, the peptide comprises the amino acid sequence "cysteine-X1-X2-X3-X4-X5-X6-cysteine". Alternatively, the disulfide bond is formed between two cysteine ​​residues, where one or both cysteine ​​residues are distal to the C-terminal (X6) and N-terminal (X1) residues of formula (I). For example, the peptide may comprise the amino acid sequence "cysteine-Y-X1-X2-X3-X4-X5-X6-cysteine" or "cysteine-X1-X2-X3-X4-X5-X6-Y-cysteine" or "cysteine-Y-X1-X2-X3-X4-X5-X6-Y-cysteine", where Y is one or more amino acid residues. In other examples, the peptide may comprise the amino acid sequence "cysteine-Y-X2-X3-X4-X5-X6-cysteine" or "cysteine-X2-X3-X4-X5-X6-Y-cysteine" or "cysteine-Y-X2-X3-X4-X5-X6-Y-cysteine", where Y is one or more amino acid residues. In one embodiment, the cyclic peptide is formed by a disulfide bond between two cysteine ​​residues.

[0049] As described elsewhere herein, the inventors have unexpectedly found that certain cyclic peptides comprising the amino acid sequence of formula (I) have higher biological activity compared to their linear counterparts.For example, the inventors have shown that the cyclic peptide CQEQLERALNSSC, cyclized by a disulfide bond between two cysteine ​​residues, has a greater binding affinity to LanCL and is more effective in vivo in an animal model of influenza A respiratory infection compared to its non-cyclized counterpart QEQLERALNSS.Thus, in one embodiment, the cyclic peptide comprises the amino acid sequence CQEQLERALNSSC.In one embodiment, the cyclic peptide consists of the amino acid sequence CQEQLERALNSSC.

[0050] The peptides described herein may be prepared by any suitable method known to those of skill in the art, illustrative examples of which include solution or solid phase synthesis using Fmoc or Boc protected amino acid residues, as well as recombinant techniques known in the art using standard microbial culture techniques, genetically engineered microorganisms and recombinant DNA techniques (Sambrook and Russell, Molecular Cloning: A Laboratory Manual (3rd ed.), 2001, CSHL Press).

[0051] In one embodiment, the peptides described herein are formed as pharma-ceutically acceptable salts. It should be understood that non-pharma-ceutically acceptable salts are also contemplated, since they may be useful as intermediates in the preparation of pharma-ceutically acceptable salts, or may be useful in storage or transportation. Suitable pharma- ceutically acceptable salts are known to those skilled in the art, and illustrative examples thereof include salts of pharma- ceutically acceptable inorganic acids (e.g., hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, boric acid, sulfamic acid, and hydrobromic acid), or salts of pharma- ceutically acceptable organic acids (e.g., acetic acid, propionic acid, butyric acid, tartaric acid, maleic acid, hydroxymaleic acid, fumaric acid, maleic acid, citric acid, lactic acid, mucic acid, gluconic acid, benzoic acid, succinic acid, oxalic acid, phenylacetic acid, methanesulfonic acid, toluenesulfonic acid, benzonesulfonic acid, salicylic acid, sulfanilic acid, aspartic acid, glutamic acid, edetic acid, stearic acid, palmitic acid, oleic acid, lauric acid, pantothenic acid, tannic acid, ascorbic acid, and valeric acid). Illustrative examples of suitable base salts include those formed with pharma- ceutically acceptable cations (e.g., sodium, potassium, lithium, calcium, magnesium, ammonium, and alkylammonium). Basic nitrogen-containing groups may be quaternized with lower alkyl halides (e.g., methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides), dialkyl sulfates (e.g., dimethyl and diethyl sulfates), and other agents.

[0052] Also disclosed herein are prodrugs comprising the peptides described herein or their pharmaceutically acceptable salts.As used herein, "prodrug" typically refers to a compound that can be metabolized in vivo to provide or release the active peptides described herein or their pharmaceutically acceptable salts.In one embodiment, the prodrug itself also shares the same or substantially the same therapeutic activity as the peptides described herein or their pharmaceutically acceptable salts, as described elsewhere herein.

[0053] In some embodiments, the peptides described herein or pharma- ceutically acceptable salts thereof may further comprise a C-terminal capping group. The term "C-terminal capping group" as used herein refers to a group that blocks the reactivity of the C-terminal carboxylic acid. A suitable C-terminal capping group forms an amide group or an ester with the C-terminal carboxylic acid, for example, the C-terminal capping group is -C(O)NHR a OR -C(O)OR b where C(O) is from the C-terminal carboxylic acid group and R a is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl or aryl; R b is alkyl, alkenyl, alkynyl, cycloalkyl, or aryl. In certain embodiments, the C-terminal capping group is -NH2, forming -C(O)NH2. In some embodiments, the peptide described herein or a pharma- ceutically acceptable salt thereof comprises a C-terminal polyethylene glycol (PEG). In one embodiment, the PEG has a molecular weight of 220-5500 Da, preferably 220-2500 Da, and more preferably 570-1100 Da.

[0054] In some embodiments, the peptides described herein or pharma- ceutically acceptable salts thereof may further comprise an N-terminal capping group. The term "N-terminal capping group" as used herein refers to a group that blocks the reactivity of the N-terminal amino group. A suitable N-terminal capping group is an acyl group that forms an amide group with the N-terminal amino group, for example, the N-terminal capping group is -NHC(O)R a where NH is from the N-terminal amino group and R a is alkyl, alkenyl, alkynyl, cycloalkyl, or aryl. In certain embodiments, the N-terminal capping group is -C(O)CH3 (acyl), forming -NHC(O)CH3.

[0055] In some embodiments, the peptides described herein or pharma- ceutically acceptable salts thereof may include C-terminal and N-terminal capping groups as described herein.It should be understood that the peptides disclosed herein do not include the full-length amino acid sequence of human growth hormone or non-human isoforms thereof.

[0056] Treatment and prevention methods As described elsewhere herein, the inventors have surprisingly found that the peptides described herein have advantageous properties that make them useful for therapeutic use, including the treatment of conditions associated with cellular aging, injury, and stress. Illustrative examples of such conditions include aging, pain, inflammatory conditions / inflammation, and microbial infections. Due to the activity observed in the peptides described herein, they are also useful as anti-aging compounds. Thus, the peptides described herein may be suitably used to treat, alleviate, or otherwise negate the severity of such conditions, including one or more symptoms thereof, in a subject in need thereof. Thus, the present disclosure extends to a method of treating a condition in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of a peptide described herein. Also provided is the use of the peptides described herein in the manufacture of a medicament for treating a condition in a subject in need thereof. Also provided is the peptide described herein for use in treating a condition in a subject in need thereof.

[0057] In one embodiment, the condition is selected from the group consisting of pain, inflammatory airway disease, microbial infection, respiratory infection, migraine, sarcopenia, impaired glucose tolerance, diabetes, obesity, metabolic diseases and conditions associated with obesity, osteoarthritis, muscle disorders, wasting diseases, aging, cachexia, anorexia, AIDS wasting syndrome, muscular dystrophies, neuromuscular diseases, amyotrophic lateral sclerosis (ALS), motor neuron diseases, diseases of the neuromuscular junction, ophthalmological conditions, conditions of the central nervous system including neurodegenerative conditions (e.g., Parkinson's disease, Alzheimer's disease), inflammatory myopathies, burns, wounds, injuries or trauma, conditions associated with elevated LDL cholesterol, conditions associated with impaired chondrocyte, proteoglycan or collagen production or quality, conditions associated with impaired formation or quality of cartilage tissue, conditions associated with impaired mass, form or function of muscle, ligament or tendon, conditions associated with inflammation, trauma or genetic abnormalities affecting muscle or connective tissue, and bone disorders.

[0058] As used herein, the terms "treating", "treatment", and the like are used interchangeably to mean to relieve, reduce, alleviate, improve or inhibit the severity of a disease or condition, including one or more symptoms thereof. As used herein, the terms "treating", "treatment", and the like are also used interchangeably to include preventing a disease or condition, including one or more symptoms thereof.

[0059] The terms "treating", "treatment" and the like also include preventing, relieving, reducing, alleviating, improving or suppressing the severity of a disease, condition and / or one or more symptoms thereof, at least for a period of time. It should be understood that the terms "treating", "treatment" and the like do not imply that a disease, condition or one or more symptoms thereof is permanently prevented, relieved, reduced, alleviated, improved or suppressed, and thus cover the temporary prevention, relief, reduction, alleviate, improvement or suppression of the severity of a disease, condition or one or more symptoms thereof.

[0060] As used herein, the term "subject" refers to a mammalian subject for whom treatment of a disease, condition, or one or more symptoms thereof is desired. Illustrative examples of suitable subjects include primates, particularly humans, companion animals (e.g., cats, dogs, etc.), working animals (e.g., horses, donkeys, etc.), livestock animals (e.g., sheep, cows, goats, pigs, etc.), laboratory test animals (e.g., rabbits, mice, rats, guinea pigs, hamsters, etc.) and domesticated wild animals (e.g., deer, dingoes, etc. in zoos and wildlife parks). In one embodiment, the subject is a human.

[0061] It should be understood that reference herein to a subject does not mean only that the subject has a disease, condition, or one or more symptoms thereof, but also includes a subject at risk of developing a disease, condition, or one or more symptoms thereof.

[0062] In one embodiment, the methods disclosed herein comprise administering to a human subject a peptide described herein, or a pharma- ceutically acceptable salt thereof.

[0063] It should be understood that the peptides described herein or their pharmaceutically acceptable salts are advantageously administered in a therapeutically effective amount. The phrase "therapeutically effective amount" typically refers to the amount necessary to achieve a desired response. It is understood by those skilled in the art that the therapeutically effective amount of peptides varies depending on several factors, illustrative examples of which include the health and physical condition of the subject being treated, the taxonomic group of the subject being treated, the severity of the disease, condition or symptom being treated, the dosage form of the composition comprising the peptides described herein or their pharmaceutically acceptable salts, the route of administration, and any combination thereof.

[0064] The therapeutically effective amount typically falls within a relatively broad range that can be determined through routine testing by those skilled in the art. Illustrative examples of suitable therapeutically effective amounts of the peptides described herein and their pharma- ceutically acceptable salts for administration to human subjects include about 0.001 mg / kg body weight to about 1 g / kg body weight, preferably about 0.001 mg / kg body weight to about 50 g / kg body weight, more preferably about 0.01 mg / kg body weight to about 1.0 mg / kg body weight.In one embodiment disclosed herein, a therapeutically effective amount of a peptide and / or a pharma- ceutically acceptable salt thereof described herein is from about 0.001 mg / kg body weight to about 1 g / kg body weight per dose (e.g., 0.001 mg / kg, 0.005 mg / kg, 0.01 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.15 mg / kg, 0.2 mg / kg, 0.25 mg / kg, 0.3 mg / kg, 0.35 mg / kg, 0.4 mg / kg, 0.45 mg / kg, 0.5 mg / kg, 0.55 mg / kg, 0.5 mg / kg, 0.6 mg / kg, 0.7 mg / kg, 0.8 mg / kg, 0.9 mg / kg, 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, 21 mg / kg, 22 mg / kg, 23 mg / kg, 24 mg / kg, 25 mg / kg, 26 mg / kg, 27 mg / kg, 28 mg / kg, 29 mg / kg, 30 mg / kg, 31 mg / kg, 32 mg / kg, 33 mg / kg, 34 mg / kg, 35 mg / kg, 36 mg / kg, 37 mg / kg, 38 mg / kg, 39 mg / kg, 40 mg / kg, 41 mg / kg, 42 mg / kg, 43 mg / kg, 44 mg / kg, 45 mg / kg, 46 mg / kg, 47 mg / kg, 48 mg / kg, 49 mg / kg, 50 mg / kg g, 0.6mg / kg, 0.65mg / kg, 0.7mg / kg, 0.75mg / kg, 0.8mg / kg, 0.85mg / kg, 0.9mg / kg, 0.95mg / kg, 1mg / kg, 1.5mg / kg, 2mg / kg, 2.5mg / kg, 3mg / kg, 3.5m g / kg, 4mg / kg, 4.5mg / kg, 5mg / kg, 5.5mg / kg, 6mg / kg, 6.5mg / kg, 7mg / kg, 7.5mg / kg, 8mg / kg, 8.5mg / kg, 9mg / kg, 9.5mg / kg, 10mg / kg, 10.5mg / kg, 11m g / kg, 11.5mg / kg, 12mg / kg, 12.5mg / kg, 13mg / kg, 13.5mg / kg, 14mg / kg, 14.5mg / kg, 15mg / kg, 15.5mg / kg, 16mg / kg, 16.5mg / kg, 17mg / kg, 17.5mg / k g, 18mg / kg, 18.5mg / kg, 19mg / kg, 19.5mg / kg, 20mg / kg, 20.5mg / kg, 21mg / kg, 21.5mg / kg, 22mg / kg, 22.5mg / kg, 23mg / kg, 23.5mg / kg, 24mg / kg, 24. 5mg / kg, 25mg / kg, 25.5mg / kg, 26mg / kg, 26.5mg / kg, 27mg / kg, 27.5mg / kg, 28mg / kg, 28.5mg / kg, 29mg / kg, 29.5mg / kg, 30mg / kg, 35mg / kg, 40mg / kg, 45mg / kg, 50mg / kg, 55mg / kg, 60mg / kg, 65mg / kg, 70mg / kg, 75mg / kg, 80mg / kg, 85mg / kg, 90mg / kg, 95mg / kg, 100mg / kg, 105mg / kg, 110mg / kg body weight, etc.).In one embodiment, the therapeutically effective amount of the peptide or pharma- ceutically acceptable salt thereof described herein is about 0.001 mg / kg body weight to about 50 mg / kg body weight. In one embodiment, the therapeutically effective amount of the peptide or pharma- ceutically acceptable salt thereof described herein is about 0.01 mg / kg body weight to about 100 mg / kg body weight. In one embodiment, the therapeutically effective amount of the peptide or pharma- ceutically acceptable salt thereof described herein is about 0.1 mg / kg body weight to about 10 mg / kg body weight, preferably about 0.1 mg / kg body weight to about 5 mg / kg body weight, more preferably about 0.1 mg / kg body weight to about 1.0 mg / kg body weight. Dosage regimens may be adjusted to obtain the optimal therapeutic response. For example, several divided doses may be administered daily, weekly, monthly or at other suitable time intervals, or the dose may be proportionally reduced as the exigencies of the situation dictate.

[0065] pain As described elsewhere herein, the inventors have found that the peptides described herein have effective analgesic properties, including the relief of neuropathic pain. Thus, in one embodiment, the condition is pain. In one embodiment, the condition is neuropathic pain.

[0066] Without being bound by theory or particular application mode, neuropathic pain is typically characterized as pain resulting from injury or disease damage to nerve tissue or nerve cells themselves, or dysfunction within nerve tissue. Pain may be peripheral, central, or a combination thereof, and in other words, the term "neuropathic pain" typically refers to any pain syndrome that is developed or caused by a primary lesion or dysfunction in the peripheral or central nervous system. Neuropathic pain is also distinguished in that it usually does not respond effectively to treatment with common analgesics such as opioids. In contrast, nociceptive pain is characterized as pain resulting from stimulation of nociceptors by noxious or potentially harmful stimuli that may cause damage or injury to tissue. Nociceptive pain usually responds to common analgesics such as opioids.

[0067] The term "analgesia" is used herein to describe a state of reduced pain perception, including loss of pain sensation, as well as a state of reduced or lost sensitivity to noxious stimuli. Such a state of reduced or lost pain sensation is typically induced by administration of a pain control agent(s) and occurs without loss of consciousness, as is commonly understood in the art. Suitable methods for determining whether a compound can provide an analgesic effect are known to those skilled in the art, illustrative examples of which include the use of animal models of neuropathic pain, such as chronic constriction injury, spinal nerve ligation and partial sciatic nerve ligation (see Bennett et al., 2003; Curr. Protoc. Neurosci., Chapter 9, Unit 9.14), and animal models of nociceptive pain, such as formalin, carrageenan or complete Freund's adjuvant (CFA)-induced inflammatory pain. Other suitable pain models are discussed by Gregory et al. (2013, J. Pain.; 14(11); An overview of animal models of pain: disease models and outcome measures).

[0068] As known to those skilled in the art, there are many possible causes of neuropathy and neuropathic pain.Therefore, it should be understood that it is contemplated herein to treat or prevent neuropathic pain regardless of its cause.In some embodiments, neuropathic pain is the result of disease or condition that affects nerves (primary neuropathy) and / or neuropathy caused by systemic disease (secondary neuropathy), and illustrative examples thereof include diabetic neuropathy; shingles-associated neuropathy; fibromyalgia; multiple sclerosis, stroke, spinal cord injury; chronic postoperative pain, phantom limb pain, Parkinson's disease; uremia-associated neuropathy; amyloidosis neuropathy; HIV sensory neuropathy; hereditary motor sensory neuropathy (HMSN); hereditary sensory neuropathy (HSN); hereditary sensory autonomic neuropathy; hereditary neuropathy with ulcer injury; nitrofurantoin neuropathy; Tomaculus neuropathy; neuropathy caused by nutritional deficiency; neuropathy caused by renal failure and complex regional pain syndrome. Other illustrative examples of conditions that can cause neuropathic pain include repetitive activities such as typing or working on an assembly line, some antiretroviral drugs such as ddC (zalcitabine) and ddI (didanosine), antibiotics (metronidazole, an antibiotic used for Crohn's disease, isoniazid, used for tuberculosis), gold compounds (used for rheumatoid arthritis), some chemotherapy drugs (such as vincristine), and many others. Compounds such as alcohol, lead, arsenic, mercury, and organophosphate pesticides are also known to cause peripheral neuropathy. Some peripheral neuropathies are associated with infectious processes (such as Guillain-Barre syndrome). Other illustrative examples of neuropathic pain include thermal or mechanical hyperalgesia, thermal or mechanical allodynia, diabetic pain, neuropathic pain affecting the oral cavity (e.g., trigeminal neuropathic pain, atypical toothache (phantom toothache), burning mouth syndrome), fibromyalgia, and entrapment pain.

[0069] In one embodiment disclosed herein, the neuropathic pain is selected from the group consisting of diabetic neuropathy; shingles-associated neuropathy; fibromyalgia; multiple sclerosis, stroke, spinal cord injury; chronic post-operative pain, phantom limb pain, Parkinson's disease; uremia-associated neuropathy; amyloidosis neuropathy; HIV sensory neuropathy; hereditary motor and sensory neuropathy (HMSN); hereditary sensory neuropathy (HSN); hereditary sensory and autonomic neuropathy; hereditary neuropathy associated with ulcer lesions; nitrofurantoin neuropathy; Tomaculus neuropathy; neuropathy caused by nutritional deficiency; neuropathy caused by renal failure; trigeminal neuropathic pain; atypical toothache (phantom toothache); burning mouth syndrome; complex regional pain syndrome; repetitive strain injury; drug-induced peripheral neuropathy; peripheral neuropathy associated with infection; allodynia; hyperesthesia; hyperalgesia; burning pain and lancinating pain.

[0070] In some embodiments, neuropathic pain may be accompanied by numbness, weakness and loss of reflexes. Pain may become severe and disabling. "Hyperalgesia" refers to an increased response to a normally painful stimulus. A state of hyperalgesia involves pain caused by a normally non-painful stimulus. The term "hyperesthesia" refers to excessive physical sensitivity, especially of the skin. The term "allodynia" as used herein refers to pain resulting from a non-noxious stimulus, i.e., pain from a stimulus that does not normally induce pain. Illustrative examples of allodynia include thermal allodynia (pain from cold or hot stimuli), tactile allodynia (pain from light pressure or touch), mechanical allodynia (pain from strong pressure or pinprick), and the like.

[0071] Neuropathic pain can be acute or chronic, and in this context, it should be understood that the time course of neuropathy can vary based on its underlying cause. For example, in cases of trauma, the onset of neuropathic pain or neuropathic pain symptoms is acute or sudden, whereas most severe symptoms develop over time and may persist for years. A chronic course over weeks to months usually indicates a toxic or metabolic neuropathy. Chronic, slowly progressive neuropathy may have a course over many years, such as painful diabetic neuropathy or most inherited neuropathies, conditions of chronic inflammatory demyelinating polyneuropathy (CIDP). Neuropathic conditions with symptoms that relapse and remitting include Guillain-Barré syndrome.

[0072] In some embodiments, the neuropathic pain results from a condition characterized by neurological hypersensitivity, such as fibromyalgia or irritable bowel syndrome.

[0073] In other embodiments, neuropathic pain results from disorders associated with abnormal nerve regeneration resulting in nerve hypersensitivity, including mastalgia, interstitial cystitis, vulvodynia, and cancer chemotherapy-induced neuropathy.

[0074] In some embodiments, the neuropathic pain is associated with surgery, pre-operative pain and post-operative pain, particularly post-operative neuropathic pain.

[0075] microbial infection Microbial infections caused by bacterial, viral and fungal pathogens remain a major global health problem with significant socio-economic losses. While treatment of bacterial infections often relies on antibiotics, the standard approach for viral infections remains supportive care and symptom relief. While such treatments have shown some success, emerging and re-emerging pathogens continue to plague human and non-human populations, at least in part as a result of mutations that give rise to novel strains with enhanced infectivity and / or resistance to existing pharmacological interventions. Lack of timely availability of antiviral drugs, including vaccines, also complicates the containment of global viral epidemics.

[0076] There are over 200 serologically known strains of viruses that cause infections, including respiratory infections, of which the most common are rhinoviruses (30–50%). Others include coronaviruses (10–15%), influenza viruses (5–15%), human parainfluenza viruses, human respiratory syncytial viruses, adenoviruses, enteroviruses and metapneumoviruses. More than 30 coronaviruses have been identified, but only three or four are known to cause respiratory infections in humans. Furthermore, coronaviruses are generally difficult to culture in vitro, making it difficult to study their function and develop appropriate treatments. Coronaviruses are enveloped, positive-stranded RNA viruses that bud from the endoplasmic reticulum-Golgi intermediate compartment or the cis-Golgi network. Coronaviruses infect humans and animals. Human coronaviruses, 229E, OC43, and the recently identified severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; see Zhu N et al., N Engl J Med., 2020), are known to be major causes of respiratory infections and can cause pneumonia, especially in the elderly, newborns, and immunocompromised individuals. Illustrative examples of coronaviruses that cause respiratory infections are described in U.S. Patent Publication No. 20190389816, the contents of which are incorporated herein by reference in their entirety.

[0077] Another widespread viral infection is caused by human rhinovirus (HRV), a member of the Enterovirus genus in the Picornaviridae family. HRV infects the upper and lower respiratory tract, including the nasal mucosa, paranasal sinuses, and middle ear, with infection causing symptoms similar to those of the common cold. Infection is usually self-limiting and restricted to the upper respiratory tract.

[0078] Some viral infections are asymptomatic in some people but contagious in others. In these cases, widespread transmission of the virus is possible because infected people do not appear sick. Transmission can be particularly harmful in places where susceptible groups live in close contact, such as schools, hospitals, and nursing homes.

[0079] Currently, only a few antiviral drugs are approved for the treatment or prevention of respiratory viral infections, including influenza or the common cold. These include oseltamivir phosphate (brand name Tamiflu®), zanamivir (brand name Relenza®), peramivir (brand name Rapivab®) and baloxavir marboxil (brand name Xofluza®). Treatment of respiratory infections is usually based on managing symptoms (e.g., sneezing, stuffy nose, rhinorrhea, eye irritation, sore throat, cough, headache, fever, chills), usually with over-the-counter oral antihistamines, aspirin, cough suppressants and nasal decongestants. Symptomatic treatment usually involves antihistamines and / or vasoconstrictor decongestants, many of which have unwanted side effects such as drowsiness.

[0080] Without being bound by theory or a particular mode of application, the inventors have surprisingly found that the peptides described herein can be used to treat microbial infections, including alleviating at least some of the symptoms of infections, such as respiratory infections.

[0081] Respiratory tract infections (RTIs) are typically defined as any infectious disease of the upper or lower respiratory tract. Upper respiratory tract infections (URTIs) include colds, laryngitis, pharyngitis / tonsillitis, acute rhinitis, acute rhinosinusitis and acute otitis media. Lower respiratory tract infections (LRTIs) include acute bronchitis, bronchitis, pneumonia and tracheitis. In primary care, antibiotics are commonly prescribed for RTIs in adults and children. RTIs account for 60% of all antibiotic prescriptions in general practice and represent a significant cost to the health system (NICE Clinical Guidelines, No. 69; Centre for Clinical Practice at NICE (UK), London: National Institute for Health and Clinical Excellence (UK); 2008).

[0082] Pathogens causing infections of the upper and / or lower respiratory tract in human and non-human subjects are known to those skilled in the art and include bacteria and viruses, illustrative examples of which are described by Charlton et al. (Clinical Microbiology Reviews; 2018, 32(1):e00042-18), Popescu et al. (Microorganisms. 2019; 7(11):521) and Kikkert, M. (J Innate Immun. 2020; 12(1):4-20), the contents of which are incorporated herein by reference in their entireties. In one embodiment, the respiratory infection is a viral infection.

[0083] Viruses that cause infections of the respiratory tract (upper and / or lower respiratory tract) in humans and non-human subjects are known to those of skill in the art, and illustrative examples thereof include picornaviruses, coronaviruses, influenza viruses, parainfluenza viruses, respiratory syncytial viruses, adenoviruses, enteroviruses, and metapneumoviruses. Thus, in one embodiment disclosed herein, the virus is selected from the group consisting of picornaviruses, coronaviruses, influenza viruses, parainfluenza viruses, respiratory syncytial viruses, adenoviruses, enteroviruses, and metapneumoviruses. In one embodiment, the virus is an influenza virus. In other embodiments, the virus is a coronavirus. Illustrative examples of coronaviruses that cause respiratory infections are known to those of skill in the art, and illustrative examples thereof include SARS-CoV-2, previously described in Zhu N et al. (N Engl J Med. 2020) and US Patent Publication No. 20190389816, the contents of which are incorporated herein by reference in their entirety. In one embodiment, the virus is SARS-CoV-2.

[0084] The peptides described herein may be particularly useful for treating respiratory infections in subjects with underlying diseases that exacerbate respiratory infections. Such underlying diseases are known to those skilled in the art, and illustrative examples thereof include chronic obstructive pulmonary disease, asthma, cystic fibrosis, emphysema and lung cancer. In one embodiment, the subject has an additional respiratory condition selected from the group consisting of chronic obstructive pulmonary disease, asthma, cystic fibrosis and lung cancer. In other embodiments, the subject is immunocompromised, whether as a result of treatment (e.g., by chemotherapy, radiation therapy) or not (e.g., by HIV infection).

[0085] Viral replication in humans usually begins 2-6 hours after initial contact. In some cases, patients are infectious for 2-3 days before the onset of symptoms. Symptoms usually begin about 2-5 days after initial infection. Respiratory infections, such as the common cold, are most contagious during the first 2-3 days after symptoms appear. There is currently no known treatment to shorten the duration of a common cold, but symptoms usually resolve spontaneously within 7-10 days, although symptoms may persist for up to 3 weeks. The virus may still be infectious until symptoms have completely resolved.

[0086] As described elsewhere herein, the inventors have also surprisingly found that the peptides described herein are effective in limiting viral replication in vivo and reducing hyperinflammation and severe cases during IAV infection.

[0087] Inflammatory Airway Disease In embodiments disclosed herein, the condition is an inflammatory airway disease. Inflammatory airway diseases, such as chronic obstructive pulmonary disease (COPD), asthma, chronic bronchitis, emphysema, cystic fibrosis, lung cancer, and bronchopulmonary dysplasia, are among the most prevalent diseases in the world. In particular, the prevalence of asthma has increased over the past two decades, and it now affects up to 10% of the population in most developed countries. COPD is the sixth leading cause of death in the world, affecting approximately 4-6% of people over the age of 45. It goes without saying that inflammatory airway diseases are a major economic burden for society, considering both direct and indirect costs.

[0088] Asthma and COPD are identified by the presence of characteristic symptoms and functional abnormalities, and airway obstruction is a prerequisite for both diseases. Airway obstruction in asthma is typically reversible, whereas COPD is typically characterized by abnormalities in expiratory flow that do not change significantly over an observation period of several months. Both airway diseases are associated with lung inflammation caused by a variety of triggers, including environmental allergens and carcinogens, occupational sensitizers, tobacco smoke, asbestos, and silica. However, it should be noted that some non-smoking asthmatics develop irreversible airway obstruction similar to that of COPD.

[0089] Chronic obstructive pulmonary disease is a growing medical problem that is expected to worsen with the aging population and increasing global use of tobacco products. Smoking cessation is the only effective preventive measure. Employers are in a unique position to assist their employees in smoking cessation. Because lung function continues to decline during long asymptomatic periods, many patients first seek medical attention at an advanced stage or when experiencing an acute exacerbation. To maintain patients' quality of life and reduce the medical costs associated with this chronic disease, clinicians need to accurately diagnose the condition and appropriately manage patients over the long term of the disease course.

[0090] As Devine, FJ (2008; Am Health Drug Benefits; 1(7):34-42) points out, COPD is a poorly reversible lung disease and one of the leading causes of morbidity and mortality worldwide. Contrary to trends in other major chronic diseases in the United States, the prevalence and mortality of COPD continues to rise, doubling between 1970 and 2002, and women now outnumber men in COPD. The majority of COPD events are caused by smoking, making it an essentially preventable disease. Most people with COPD are middle-aged or older. Few effective treatments for COPD have been found. The only measure known to reduce disease development is smoking cessation.

[0091] Asthma is a diverse and multifactorial disease with variable and mostly reversible obstruction of the respiratory pathways based on a chronic bronchial inflammatory response (Horak et al., 2016; Wien Klin Wochenschr., 128(15):541-554). Asthma symptoms (cough, phlegm, nasal discharge, wheezing, chest tightness or shortness of breath) are variable and generally correlate with expiratory flow limitation. Due to its diversity, many different phenotypes are ascribed to asthma, including allergic asthma, nonallergic asthma, childhood asthma / recurrent obstructive bronchitis, late-onset asthma, asthma with fixed airflow obstruction, obesity-related asthma, occupational asthma, asthma in the elderly and severe asthma.

[0092] Asthma treatment (pharmacological and non-pharmacological interventions) is mainly based on symptom management (cycles of assessment, adjustment and reassessment) and is usually associated with suppression of asthma exacerbations. From a pharmacological point of view, the gold standard of asthma treatment is generally low-dose inhaled corticosteroids, often combined with on-demand short-acting beta-2-agonists (SABAs). Other treatments include LTRAs (leukotriene receptor antagonists), combinations of low-dose inhaled corticosteroids and long-acting beta-2-agonists (LABAs). However, existing treatments can cause side effects, especially during long-term use. Common side effects of preventive medicines (e.g., inhaled corticosteroids) are hoarseness, sore mouth and throat, and fungal infections of the throat.

[0093] The inventors have surprisingly found that the peptides described herein are capable of ameliorating at least some of the inflammatory mediators of inflammatory airway diseases.

[0094] Inflammatory airway diseases are known to those skilled in the art, and illustrative examples thereof include chronic obstructive pulmonary disease (COPD), asthma, chronic bronchitis, emphysema, cystic fibrosis, lung cancer, and bronchopulmonary dysplasia. In one embodiment, the inflammatory airway disease is COPD. In one embodiment, the inflammatory airway disease is asthma. In one embodiment, the inflammatory airway disease is chronic bronchitis. In one embodiment, the inflammatory airway disease is emphysema. In one embodiment, the inflammatory airway disease is cystic fibrosis. In one embodiment, the inflammatory airway disease is associated with lung cancer. In one embodiment, the inflammatory airway disease is bronchopulmonary dysplasia.

[0095] The methods described herein may be particularly useful for treating inflammatory airway disease in subjects susceptible to diseases that exacerbate inflammatory airway disease.Such underlying diseases are known to those skilled in the art, and illustrative examples thereof include, for example, respiratory infections caused by viruses, bacteria or other pathogens.In other embodiments, the subject is immunocompromised, whether as a result of treatment (e.g., by chemotherapy, radiotherapy) or not (e.g., by HIV infection).

[0096] Route of administration The peptides and pharmaceutically acceptable salts thereof as described herein may be administered to a subject by any suitable route that allows delivery of a therapeutically effective amount of the peptide or pharmaceutically acceptable salt thereof to the subject as described herein. Suitable administration routes are known to those skilled in the art, and illustrative examples thereof include enteral administration routes (e.g., oral and rectal administration), parenteral administration routes, typically by injection or microinjection (e.g., intramuscular, subcutaneous, intravenous, epidural, intraarticular, intraperitoneal, intracisternal or intrathecal administration), and local (transdermal or transmucosal) administration routes (e.g., buccal, sublingual, intravaginal, intranasal, or by inhalation, insufflation, suppository or nebulization). In one embodiment, the administration route is by inhalation or insufflation. The peptides and pharmaceutically acceptable salts thereof as described herein may also be administered to a subject suitably as sustained release dosage forms to provide sustained release of the active ingredient(s) over an extended period of time. The term "controlled release" typically refers to a release of an active ingredient to provide a constant or substantially constant concentration of the active ingredient in a subject over a period of time (e.g., from about 8 hours up to about 12 hours, up to about 14 hours, up to about 16 hours, up to about 18 hours, up to about 20 hours, up to a day, up to a week, up to a month, or more than a month). Controlled release of the active ingredient can optionally begin within minutes of administration or after a lag time after administration. Suitable controlled release dosage forms are known to those skilled in the art, and illustrative examples thereof are described by Anal, AK (2010; Controlled-Release Dosage Forms; Pharmaceutical Sciences Encyclopedia; 11:1-46).

[0097] Without being bound by theory or a particular mode of application, it may be desirable to select a route of administration based on the severity of the disease, condition, or one or more symptoms thereof, as described herein. In one embodiment disclosed herein, the peptides described herein or pharma- ceutically acceptable salts thereof are administered enterally to the subject. In one embodiment disclosed herein, the peptides described herein or pharma- ceutically acceptable salts thereof are administered orally to the subject. In one embodiment disclosed herein, the peptides described herein or pharma- ceutically acceptable salts thereof are administered parenterally to the subject. In other embodiments disclosed herein, the peptides described herein or pharma- ceutically acceptable salts thereof are administered topically to the subject. In other embodiments disclosed herein, the peptides described herein or pharma- ceutically acceptable salts thereof are administered to the subject by inhalation. In other embodiments disclosed herein, the peptides described herein or pharma- ceutically acceptable salts thereof are administered to the subject by insufflation.

[0098] As described elsewhere herein, "topical" administration typically refers to application of an active ingredient to a surface of the body, such as the skin or mucosa, preferably in the form of a cream, lotion, foam, gel, ointment, nose drop, eye drop, ear drop, transdermal patch, transdermal film (e.g., sublingual film), and the like. Topical administration also includes administration via the mucous membranes of the respiratory tract by inhalation or insufflation. In one embodiment disclosed herein, topical administration is selected from the group consisting of transdermal administration and transmucosal administration. In one embodiment, the peptide or pharma- ceutically acceptable salt thereof described herein is administered to a subject transdermally. In one embodiment, the peptide or pharma- ceutically acceptable salt thereof described herein is administered to a subject by inhalation, insufflation, or nebulization.

[0099] In one embodiment, the method comprises administering a peptide described herein, or a pharma- ceutically acceptable salt thereof, to a human by inhalation or insufflation. In other embodiments, the method comprises administering a peptide described herein, or a pharma- ceutically acceptable salt thereof, to a non-human subject by inhalation or insufflation. In yet other embodiments, the method comprises administering a peptide described herein, or a pharma- ceutically acceptable salt thereof, to a non-human subject selected from the group consisting of a cat, a dog, and a horse.

[0100] In one embodiment, the method comprises orally administering a peptide described herein or a pharma- ceutically acceptable salt thereof to a human. In other embodiments, the method comprises orally administering a peptide described herein or a pharma- ceutically acceptable salt thereof to a non-human subject. In yet other embodiments, the method comprises orally administering a peptide described herein or a pharma- ceutically acceptable salt thereof to a non-human subject selected from the group consisting of a cat, a dog, and a horse.

[0101] Illustrative examples of topical administration are described elsewhere herein. In one embodiment, the topical administration is transdermal administration.

[0102] In one embodiment disclosed herein, the peptides described herein or pharma- ceutically acceptable salts thereof are administered to a subject as a sustained release dosage form, illustrative examples of which are described elsewhere herein. In one embodiment, the method comprises administering the peptides described herein or pharma- ceutically acceptable salts thereof to a human as a sustained release dosage form. In other embodiments, the method comprises administering the peptides described herein or pharma- ceutically acceptable salts thereof to a non-human subject as a sustained release dosage form. In yet other embodiments, the method comprises administering the peptides described herein or pharma- ceutically acceptable salts thereof to a non-human subject selected from the group consisting of cats, dogs, and horses as a sustained release dosage form.

[0103] As described elsewhere herein, several (i.e., multiple) divided doses may be administered daily, weekly, monthly or at other suitable time intervals, or the dose may be proportionally reduced depending on the exigencies of the situation. When multiple doses of therapeutic course are necessary or desired, it may be beneficial to administer the peptides disclosed herein via more than one route. For example, it may be desirable to administer a first dose parenterally (e.g., via intramuscular, intravenous, subcutaneous, epidural, intraarticular, intraperitoneal, intracisternal or intrathecal administration routes) to induce a rapid or acute therapeutic effect in the subject, and then administer subsequent (e.g., second, third, fourth, fifth, etc.) doses enterally (e.g., orally or rectally), by inhalation or insufflation, and / or topically (e.g., via transdermal or transmucosal administration routes) to ensure sustained availability of the active ingredient over an extended period following the acute phase of treatment. Alternatively, it may be desirable to administer a dose enterally (e.g., orally or rectally) and then administer subsequent (e.g., second, third, fourth, fifth, etc.) doses parenterally (e.g., via intramuscular, intravenous, subcutaneous, epidural, intraarticular, intraperitoneal, intracisternal or intrathecal routes of administration), by inhalation or by insufflation, and / or topically (e.g., via transdermal or transmucosal routes of administration). Alternatively, it may be desirable to administer a dose topically (e.g., via transdermal or transmucosal routes of administration) and then administer subsequent (e.g., second, third, fourth, fifth, etc.) doses parenterally (e.g., via intramuscular, intravenous, subcutaneous, epidural, intraarticular, intraperitoneal, intracisternal or intrathecal routes of administration), by inhalation or by insufflation, and / or enterally (e.g., orally or rectally).

[0104] It should also be understood that any combination of two or more administration routes may be used according to the methods disclosed herein if multiple administration routes are desired. Illustrative examples of suitable combinations include, but are not limited to, (in order of administration): (a) parenteral-enteral; (b) parenteral-topical; (c) parenteral-enteral-topical; (d) parenteral-topical-enteral; (e) enteral-parenteral; (f) enteral-topical; (g) enteral-topical-parenteral; (h) enteral-parenteral-topical; (i) topical-parenteral; (j) topical-enteral; (k) topical-parenteral-enteral; (l) topical-enteral-parenteral; (m) parenteral-enteral-topical-parenteral; (n) parenteral-enteral-topical-enteral, etc.

[0105] Pharmaceutical Compositions The peptides described herein or pharma- ceutically acceptable salts thereof may be formulated for administration to a subject as a neat chemical. However, in certain embodiments, it may be preferred to formulate the peptides described herein or pharma- ceutically acceptable salts thereof as pharmaceutical compositions, including veterinary compositions. Thus, in other aspects disclosed herein, there is provided a peptide described herein for use in treating a condition in a subject in need thereof, as described herein.

[0106] As described elsewhere herein, the peptides and pharma- ceutically acceptable salts thereof described herein may be administered with one or more other active ingredients suitable for the underlying disease to be treated, either sequentially or in combination (e.g., as a combination drug). For example, the compositions disclosed herein may be formulated for administration with inhaled corticosteroids commonly used to treat asthma, either sequentially or in combination (e.g., as a combination drug). Other suitable combination or adjunctive therapies are known to those skilled in the art, and the choice will depend on the underlying disease or its symptoms.

[0107] In one embodiment, the composition further comprises a pharma- ceutically acceptable carrier, excipient, or diluent, as described elsewhere herein.

[0108] The peptides and pharma- ceutically acceptable salts thereof described herein may be suitably prepared as pharmaceutical compositions or unit dosage forms for use as solids (e.g., tablets or filled capsules) or liquids (e.g., solutions, suspensions, emulsions, elixirs, or filled capsules) for oral use; in the form of ointments, suppositories, or enemas for rectal administration; in the form of sterile injectable solutions for parenteral use (e.g., intramuscular, subcutaneous, intravenous, epidural, intraarticular, and intrathecal administration); or in the form of ointments, lotions, creams, gels, patches, sublingual strips, or films, etc., for parenteral (e.g., topical, buccal, sublingual, vaginal) administration. In one embodiment, the peptides and pharma- ceutically acceptable salts thereof described herein are formulated for topical (e.g., transdermal) delivery. Suitable transdermal delivery systems are known to those of skill in the art, and illustrative examples thereof are described by Prausnitz and Langer (2008; Nature Biotechnol.; 26(11):1261-1268), the contents of which are incorporated herein by reference. In other embodiments, the peptides and pharma- ceutically acceptable salts thereof described herein are formulated for sublingual or buccal delivery. Suitable sublingual and buccal delivery systems are known to those of skill in the art, and illustrative examples thereof are described by Bala et al. (2013; Int. J. Pharm. Investig.; 3(2):67-76), the contents of which are incorporated herein by reference.

[0109] Suitable pharmaceutical compositions and unit dosage forms thereof may contain conventional ingredients in conventional proportions, with or without additional active compounds or ingredients, and such unit dosage forms may contain any suitable effective amount of active ingredient consistent with the intended daily dose range. The peptides described herein and their pharmaceutically acceptable salts may be formulated for administration in a wide variety of enteral, topical and / or parenteral dosage forms. Suitable dosage forms may contain, as active ingredients, a combination of two or more of the peptides described herein or their pharmaceutically acceptable salts.

[0110] In one embodiment, the composition is formulated for oral administration to a human. In other embodiments, the composition is formulated for oral administration to a non-human subject. In yet other embodiments, the composition is formulated for oral administration to a non-human subject selected from the group consisting of cats, dogs and horses.

[0111] In other embodiments, the composition is formulated for parenteral administration to a human. In other embodiments, the composition is formulated for parenteral administration to a non-human subject. In yet other embodiments, the composition is formulated for parenteral administration to a non-human subject selected from the group consisting of cats, dogs, and horses. In one embodiment, the parenteral administration is subcutaneous administration.

[0112] In other embodiments, the composition is formulated for topical administration to a human. In other embodiments, the composition is formulated for topical administration to a non-human subject. In yet other embodiments, the composition is formulated for topical administration to a non-human subject selected from the group consisting of cats, dogs and horses. In one embodiment, the topical administration is transdermal.

[0113] In other embodiments, the composition is formulated for administration by inhalation or insufflation to a human. In other embodiments, the composition is formulated for administration by inhalation or insufflation to a non-human subject. In yet other embodiments, the composition is formulated for administration by inhalation or insufflation to a non-human subject selected from the group consisting of cats, dogs and horses.

[0114] In other embodiments, the composition is formulated as a sustained release dosage form for administration to a human. In other embodiments, the composition is formulated as a sustained release dosage form for administration to a non-human subject. In yet other embodiments, the composition is formulated as a sustained release dosage form for administration to a non-human subject selected from the group consisting of cats, dogs and horses. Illustrative examples of suitable sustained release dosage forms are described elsewhere herein.

[0115] For preparing the pharmaceutical compositions described herein, pharma- ceutically acceptable carriers can be either solid or liquid. Illustrative examples of solid form preparations include powders, tablets, pills, capsules, cachets, suppositories and dispersible granules. A solid carrier can be one or more substances that can also act as diluents, flavorings, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents or encapsulating materials. In the case of powders, the carrier can be a finely divided solid that is in a mixture with the finely divided active ingredient. In the case of tablets, the active ingredient can be mixed with a carrier having the necessary binding capacity in a suitable ratio and compressed into the desired shape and size.

[0116] In some embodiments, powders and tablets contain from 5 or 10% to about 70% of the active compound. Illustrative examples of suitable carriers include magnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low melting wax, cocoa butter, and the like. The term "preparation" is intended to include a formulation of the active compound with an encapsulating material, providing a capsule in which the active ingredient is surrounded by a carrier, with or without a carrier. Similarly, cachets and lozenges are contemplated herein. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid forms suitable for oral administration.

[0117] To prepare suppositories, a low melting wax, such as fatty acid glycerides or cocoa butter, is first melted and stirred to disperse the active ingredient homogeneously therein, The molten homogeneous mixture is then poured into suitable sized molds and allowed to cool and solidify.

[0118] Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or sprays containing, in addition to the active ingredient, such carriers as are known in the art to be appropriate.

[0119] Liquid form preparations include solutions, suspensions and emulsions, for example, water or water-propylene glycol solutions.For example, parenteral injection liquid preparations can be formulated as liquid solutions, such as aqueous polyethylene glycol solutions.

[0120] The peptides described herein and their pharma- ceutically acceptable salts can be formulated for parenteral administration (e.g., injection, e.g., bolus injection or continuous infusion), and can be presented in unit dosage form in ampoules, prefilled syringes, small-volume infusions, or multi-dose containers with added preservatives.The compositions can take the form of suspensions, solutions, or emulsions in oily or aqueous vehicles, and can contain formulating agents such as suspending, stabilizing, and / or dispersing agents.Alternatively, the active compound(s) can be in the form of powder obtained by aseptic isolation of sterile solid or lyophilization from solution, to be constituted with a suitable vehicle, e.g., sterile pyrogen-free water, before use.

[0121] Aqueous solutions suitable for oral use can be prepared by dissolving the active component in water and adding suitable colorants, flavors, stabilizing, and thickening agents as desired.

[0122] Aqueous suspensions suitable for oral use may be prepared by dispersing the finely divided active ingredient in water with a viscous material such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose or other known suspending agents.

[0123] Also contemplated herein is the solid form preparation that is intended to be converted immediately before use into the liquid form preparation for oral administration.Such liquid form preparations include solutions, suspensions and emulsions.These preparations may contain, in addition to the active ingredient, coloring agents, flavoring agents, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizers, etc.

[0124] For topical administration to epidermis, the peptides described herein and their pharma- ceutically acceptable salts can be formulated as ointments, creams or lotions, or as transdermal patches.Ointments and creams can be formulated, for example, with an aqueous or oily base, with suitable thickening and / or gelling agents added.Lotions can be formulated with an aqueous or oily base, and generally also contain one or more emulsifiers, stabilizers, dispersants, suspending agents, thickening agents or coloring agents.

[0125] Formulations suitable for topical administration to the mouth include lozenges comprising the active ingredient in a flavored base, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert base such as gelatin and glycerin or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

[0126] The solution or suspension is directly applied to the nasal cavity by conventional means, for example, by using a dropper, pipette or spray.The formulation can be provided in single-dose or multi-dose form.In the latter case of a dropper or pipette, this can be achieved by the patient administering an appropriate predetermined volume of the solution or suspension.In the case of a spray, this can be achieved, for example, by a metered-dose spray pump or an inhaler.To improve nasal delivery and retention, the peptide used in the present invention can be encapsulated with cyclodextrin or formulated with an agent that is expected to enhance delivery and retention to the nasal mucosa.

[0127] Administration to the respiratory tract may also be accomplished by an aerosol formulation in which the active ingredient is provided in a pressurized pack with a suitable propellant including a chlorofluorocarbon (CFC) (e.g., dichlorodifluoromethane, trichlorofluoromethane, or dichlorotetrafluoroethane), carbon dioxide, or other suitable gas. The aerosol may conveniently also contain a surfactant such as lecithin. The dose of drug may be controlled by providing a metered valve.

[0128] Alternatively or additionally, the active ingredient can be provided in the form of a dry powder, for example a powder mixture of the compound in a suitable powder base such as lactose, starch, starch derivatives such as hydroxypropylmethylcellulose and polyvinylpyrrolidone (PVP). Conveniently, the powder carrier forms a gel in the nasal cavity. The powder composition can be presented in unit dose form, for example in capsules or cartridges of gelatin or the like, or blister packs from which the powder can be administered by an inhaler.

[0129] In formulations intended for administration to the respiratory tract, including intranasal dosage forms, the peptides will generally have a small particle size, for example of the order of 1-10 microns or less, which may be obtained by means known in the art, for example by micronization.

[0130] When desired, and as described elsewhere herein, formulations adapted to give slow or sustained release of the active ingredient may be employed.

[0131] In one embodiment, the pharmaceutical preparations described herein are preferably in unit dosage form.In such form, the preparation is subdivided into unit doses containing a suitable amount of active ingredient.The unit dosage form may be a packaged preparation, the package may contain discrete amounts of the preparation (e.g., packaged tablets, capsules, powders in vials or ampoules).The unit dosage form may also be a capsule, tablet, cachet or lozenge itself, or the appropriate number of any of these in packaged form.

[0132] In one embodiment, the compositions disclosed herein are formulated for oral administration to humans. In yet other embodiments, the compositions disclosed herein are formulated for oral administration to non-humans. In a further embodiment, the compositions disclosed herein are formulated for oral administration to non-humans selected from the group consisting of cats, dogs and horses.

[0133] In one embodiment, the compositions disclosed herein are formulated for administration by inhalation or insufflation to a human. In yet other embodiments, the compositions disclosed herein are formulated for administration by inhalation or insufflation to a non-human. In a further embodiment, the compositions disclosed herein are formulated for administration by inhalation or insufflation to a non-human selected from the group consisting of cats, dogs and horses.

[0134] In other embodiments, the peptides and pharma- ceutically acceptable salts thereof described herein are formulated for oral administration to a human subject. In other embodiments, the peptides and pharma- ceutically acceptable salts thereof described herein are formulated for oral administration to a non-human subject. In yet other embodiments, the peptides and pharma- ceutically acceptable salts thereof described herein are formulated for oral administration to a non-human subject selected from the group consisting of cats, dogs, and horses.

[0135] In other embodiments, the peptides and pharma- ceutically acceptable salts thereof described herein are formulated for topical administration to a human subject. In still other embodiments, the peptides and pharma- ceutically acceptable salts thereof described herein are formulated for topical administration to a non-human subject. In other embodiments, the peptides and pharma- ceutically acceptable salts thereof described herein are formulated for topical administration to a non-human subject selected from the group consisting of cats, dogs, and horses. In one embodiment, the topical administration is transdermal.

[0136] In other embodiments, the peptides and pharma- ceutically acceptable salts thereof described herein are formulated for administration by inhalation or insufflation to a human subject. In yet other embodiments, the peptides and pharma- ceutically acceptable salts thereof described herein are formulated for administration by inhalation or insufflation to a non-human subject. In other embodiments, the peptides and pharma- ceutically acceptable salts thereof described herein are formulated for administration by inhalation or insufflation to a non-human subject selected from the group consisting of cats, dogs, and horses.

[0137] In other embodiments, the peptides and pharma- ceutically acceptable salts thereof described herein are formulated for administration to a human subject as a sustained release dosage form. In yet other embodiments, the peptides and pharma- ceutically acceptable salts thereof described herein are formulated for administration to a non-human subject as a sustained release dosage form. In other embodiments, the peptides and pharma- ceutically acceptable salts thereof described herein are formulated for administration to a non-human subject as a sustained release dosage form, wherein the non-human subject is selected from the group consisting of cats, dogs, and horses. In one embodiment, the sustained release dosage form is formulated for parenteral administration.

[0138] As described elsewhere herein, several (i.e., multiple) divided doses may be administered daily, weekly, monthly or at other suitable time intervals, or the dose may be proportionally reduced depending on the exigencies of the situation. When multiple doses of therapeutic course are necessary or desired, the compositions disclosed herein may be suitably formulated for administration via said multiple routes. For example, it may be desirable to administer a first dose parenterally (e.g., intramuscularly, intravenously, subcutaneously, etc.) to induce a rapid or acute therapeutic effect in the subject, and then administer subsequent (e.g., second, third, fourth, fifth, etc.) doses parenterally (e.g., enterally and / or topically) to provide sustained availability of the active ingredient for an extended period following the acute phase of treatment. Thus, in one embodiment, the peptides and compositions disclosed herein are formulated for parenteral administration to a subject (i.e., as a parenteral dosage form) as a first dose, and formulated for non-parenteral administration to a subject (e.g., as an enteral and / or topical dosage form) after the first dose. In one embodiment, the parenteral administration is selected from the group consisting of intramuscular administration, subcutaneous administration, and intravenous administration, hi a further embodiment, the parenteral administration is subcutaneous administration.

[0139] In other embodiments, enteral administration is oral administration.Therefore, in one embodiment, the peptide and composition disclosed herein is formulated for parenteral administration to subject as a first dose, and after the first dose, is formulated for oral administration to subject (i.e., as oral dosage form).

[0140] In other embodiments, enteral administration is topical administration.Thus, in one embodiment, the peptide and composition disclosed herein is formulated for parenteral administration to subject as a first dose, and after the first dose, is formulated for topical administration to subject (i.e., as an oral dosage form).In one embodiment, topical administration is transdermal administration.

[0141] In other embodiments, it may be desirable to administer a first dose parenterally (e.g., intramuscularly, intravenously, subcutaneously, etc.) to induce a rapid or acute therapeutic effect in the subject, followed by subsequent administration (e.g., second, third, fourth, fifth, etc.) of a sustained release dosage form as described elsewhere herein to provide sustained release of the active ingredient over an extended period following the acute phase of treatment. Thus, in other embodiments, the peptides and compositions disclosed herein are formulated for parenteral administration to a subject as a first dose, and then formulated as a sustained release dosage form that is administered to the subject after the first dose. In one embodiment, the sustained release dosage form is formulated for parenteral administration.

[0142] It may also be desirable to administer a first dose enterally (e.g., orally or rectally) and then administer subsequent doses (e.g., second, third, fourth, fifth, etc.) topically (e.g., transdermally). Thus, in one embodiment, the peptides and compositions disclosed herein are formulated for enteral administration to a subject (i.e., as an enteral (oral or rectal) dosage form) as a first dose, and are formulated for topical administration to a subject (e.g., as a transdermal or transmucosal dosage form) after the first dose. In other embodiments, the peptides and compositions disclosed herein are formulated for topical administration selected from the group consisting of transdermal and transmucosal administration. In further embodiments, the peptides and compositions disclosed herein are formulated for transdermal administration.

[0143] In yet other embodiments, it may be desirable to administer the peptides or compositions disclosed herein enterally (e.g., orally or rectally) as a first dose, as described elsewhere herein, and then administer subsequent doses (e.g., second, third, fourth, fifth, etc.) as sustained release dosage forms. Thus, in one embodiment, the peptides and compositions disclosed herein are formulated for enteral administration as a first dose and formulated for administration as sustained release dosage forms, where the sustained release dosage forms are formulated for administration subsequent to the first dose. In one embodiment, the enterally administered dose is formulated for oral administration. In other embodiments, the sustained release dosage forms are formulated for parenteral administration.

[0144] In one embodiment, it may be desirable to administer the peptide or composition disclosed herein locally (e.g., orally or rectally) as a first dose, as described elsewhere herein, and then administer subsequent doses (e.g., second, third, fourth, fifth, etc.) as sustained release dosage forms. Thus, in one embodiment, the peptides and compositions disclosed herein are formulated for local administration as a first dose, and formulated for administration as sustained release dosage forms, where the sustained release dosage forms are formulated for administration subsequent to the first local administration dose. In one embodiment, the local administration dose is formulated for transdermal administration. In other embodiments, the sustained release dosage forms are formulated for parenteral administration.

[0145] The invention will now be described with reference to the following examples which illustrate certain preferred aspects of the invention, however it is to be understood that the specificity of the following description of the invention does not supersede the generality of the preceding description of the invention. EXAMPLES

[0146] Example 1: LanCL binding assay Gel-based analysis of cross-linked proteins Dried pellets of LANCL1 pre-photolabeled with photoprobes in the presence of different peptides or PBS / DMSO vehicle were resuspended in 30 μL of SDS loading buffer (Bio Rad's XT Sample Buffer with 2.5% v / v 2-mercaptoethanol) and heated (60 °C, 30 min). Proteins were separated using SDS-PAGE (4-15% Criterion™ TGX Stain-Free™ Protein Gel, Bio Rad) and analyzed by in-gel fluorescence scanning using a ChemiDoc™ MP Imaging System (Bio Rad) with a green LED light as the excitation source and a BP600 / 20 nm emission filter. After in-gel fluorescence scanning, gels were stained with Coomassie blue to ensure equal amounts of protein samples were loaded in each lane and imaged on the ChemiDoc™ MP Imaging System. Photoincorporation of each photoprobe into LANCL1 was quantitatively assessed by measuring the fluorescence intensity of the corresponding gel band using Image lab software (Bio Rad) and normalizing this value to the intensity value of the Coomassie blue-stained LANCL1 gel band to adjust for loading differences.

[0147] result As shown in Table 2, the peptide was found to specifically bind to LanCL1 and displace the known LanCL1 ligand, PAL-CRSVEGSCGF (SEQ ID NO: 22), from recombinant LanCL1 (rLanCL1). 50 The substitution values ​​are shown in Table 2. Note that the cyclized peptide of SEQ ID NO: 38 had an unexpectedly greater binding affinity to rLanCL1 compared to its linear counterpart, SEQ ID NO: 37. Similarly, the cyclized peptide of SEQ ID NO: 40 had a better binding affinity to rLanCL1 compared to its linear counterpart, SEQ ID NO: 39.

[0148] [Table 2]

[0149] Example 2: Respiratory epithelial cell viability By interacting with LANCL1, the peptides described in this patent have been shown to play a role in protecting cells from the harmful effects of chemical or oxidative stress. An assay was developed in which cells were stressed with a fixed dose of the chemotherapeutic drug taxol, resulting in a 50% inhibition of cell viability compared to untreated cells. The peptides were then added in increasing concentrations to the cell culture medium and assessed for their ability to restore viability to taxol-treated cells.

[0150] Briefly, A549 cells were cultured at 50000 A549 cells / well in opaque-walled multiwell plates in 100 μL of culture medium per well of a 96-well plate (DMEM medium 11960-044 Thermoscientific, +10% FBS 10270-106 Gibco, Thermoscientific, +1% Na Pyruvate S8636-100ML, Sigma, +1% Glutamax 35050061, Thermoscientific, +1% Penicillin-Streptomycin 11074440001, Sigma). Control wells containing medium without cells were used to obtain values ​​for background luminescence. Cells were incubated overnight at 37°C in 5% CO2.

[0151] Taxol (T7402-5MG, Sigma-Aldrich) was added to each well as a 10 mM solution in DMSO at a final concentration of 350 μM, which gives a 50% inhibition of proliferation compared to vehicle alone. 100 μL of medium + DMSO + peptide or medium + taxol + peptide (each concentration) was added to each well and incubated at 37° C., 5% CO2 for 16 hours.

[0152] Cell morphology, viability, and confluency were assessed by phase contrast microscopy. The number of metabolically active cells was then quantified using the CellTiter-Glo® Luminescent Cell Viability Assay (G7571, Promega - a homogeneous method that determines the number of viable cells in culture based on the quantification of ATP present) according to the manufacturer's instructions. A 100 μL volume of CellTiter-Glo® reagent was added to the 100 μL volume of cell culture medium present in each well, the contents were mixed on an orbital shaker for 2 minutes to induce cell lysis, and the plate was incubated at room temperature for 10 minutes to stabilize the luminescence signal, after which luminescence was recorded using a CLARIOstar multiwell luminometer (BMG Labtech) with an integration time of 0.5 seconds.

[0153] result As shown in Table 3, the peptides were found to restore viability of A549 cells treated in vitro with a dose of taxol that reduced proliferation by 50% compared to untreated cells. Consistent with the LanCL1 binding data in Table 2 above, the cyclized peptide of SEQ ID NO: 38 was found to restore A549 viability, but its linear counterpart SEQ ID NO: 37 did not. Similarly, the cyclized peptide of SEQ ID NO: 10 was found to restore A549 viability, but its linear counterpart SEQ ID NO: 11 did not. Unexpectedly, relatively short peptides of 3-, 4-, 5- and 6-amino acid length (SEQ ID NOs: 39, 42 and 59-61) were also found to partially restore A549 viability. The peptide of SEQ ID NO: 40 (a cyclized variant of SEQ ID NO: 39) also restored A549 viability. The peptide of SEQ ID NO:9 (a linear fragment of SEQ ID NO:1) also rescued taxol-induced loss of cell viability (see also FIG. 2).

[0154] To assess whether the effect of the peptides was dependent on LanCL1 expression, A549 cells were treated with LanCL1 siRNA (100 nM) for 48 h to knock down LanCL1 expression. The cells were then treated with taxol (IC 50 The cells were incubated in the presence of 1, 5, 25, 50 and 100 μM of either vehicle alone (dimethyl sulfoxide; DMSO) or peptide of SEQ ID NO: 1 (diluted in DMSO) in the presence of 100 μM of taxol (approximately 350 μM). Transfection with control siRNA (SiCTL) or siRNA against LanCL1 (SiLanCL1) did not alter the viability of A549 cells. As shown in Figure 1, peptide of SEQ ID NO: 1 did not significantly affect the viability of non-transfected A549 cells (NT) or A549 cells transfected with SiCTL in the absence of taxol. In cells transfected with SiLanCL1, peptide of SEQ ID NO: 1 inhibited A549 proliferation at higher doses.

[0155] In the presence of 350 μM taxol, the presence of peptide SEQ ID NO: 1 rescued the loss of viability of non-transfected A549 cells (NT) or A549 cells transfected with SiCTL. This effect represents a protective effect on epithelial cells. In contrast, peptide SEQ ID NO: 1 did not rescue the negative effect of taxol on A549 viability.

[0156] These data indicate that peptides comprising the amino acid sequence of formula (I) are able to rescue the negative effects of taxol-induced stress on epithelial cell viability, and that this rescue effect is dependent on LanCL1.

[0157] [Table 3]

[0158] Example 3: Influenza A Infection Mouse Model Six- to eight-week-old C57BL / 6 male mice were housed in the Specific Pathogen Free Physical Containment Level 2 (PC2) Animal Research Facility at Monash Medical Centre. All experimental procedures were approved by the Hudson Animal Ethics Committee and were performed in accordance with the approved guidelines. The IAV strain used in this study was HKx31 (H3N2), a high-yield reassortant of A / PR / 8 / 34 (H1N1) that retains the surface glycoproteins of A / Aichi / 2 / 1968 (H3N2). HKx31 was propagated in 10-day-old embryonated chicken eggs by standard procedures and titrated on Madin-Darby Canine Kidney (MDCK) cells.

[0159] For the viral infection study, groups of eight male C57BL / 6 mice were randomized. Mice were lightly anesthetized and given 10 5 Mice were infected intranasally with PFU of HKx31(H3N2), previously shown to induce severe disease (Rosli et al., 2019; Tate et al., 2016). Mice were treated with peptides described herein (5 or 20 mg / kg; as indicated) via the intranasal route at the indicated time points. Control mice were treated with PBS only. Mice were weighed daily and assessed for visual signs of clinical disease, including lethargy, ruffled fur, labored breathing, and hunched behavior. Animals that lost more than 20% of their original weight or showed severe clinical symptoms of disease were euthanized. Bronchoalveolar lavage (BAL) fluid was obtained immediately after euthanasia by flushing the lungs three times with 1 mL of PBS. Lungs were then excised and immediately frozen in liquid nitrogen. Titers of infectious virus in lung homogenates were measured by standard plaque assay using MDCK cells.

[0160] Quantification of cytokines in mouse BAL fluid and serum For cytokine detection, BAL fluid was collected and stored at -80°C. IL-6, MCP-1 / CCL2, IFNγ, IL-10, IL-12p70 and TNFα protein levels were determined by cytokine bead array (CBA) using a mouse inflammation kit (Becton Dickinson). Mouse IFNα levels were determined by sandwich ELISA using mouse monoclonal clone F18 (Thermo Scientific) and rabbit polyclonal antibody (PBL) (Thomas et al., 2014). Mouse IFNβ levels were determined by sandwich ELISA using mouse monoclonal clone 7F-D3 (Abcam) and rabbit polyclonal antibody (PBL) (Thomas et al., 2014). Mouse IFNλ 2 / 3 was quantified by ELISA (R&D Systems).

[0161] Recovery and characterization of leukocytes from mice For flow cytometry analysis, BAL cells were treated with red blood cell lysis buffer (Sigma Aldrich) and cell number and viability were assessed by trypan blue exclusion using a hemocytometer. BAL cells were incubated with Fc block (2.4G2; eBiosciences) and then analyzed for Ly6C, Ly6G, CD11c and IA. b The cells were stained with a fluorochrome-labeled monoclonal antibody against MHC-II (BD Biosciences, USA). Neutrophils (Ly6G + ), macrophages (CD11c + IA b低 ), dendritic cells (DC; CD11c + IA b高 ), inflammatory macrophages (Ly6G-Ly6C + ) were quantified by flow cytometry as previously described ( Rosli et al., 2019 ; Tate et al., 2016 ). Live cells (propidium iodide negative) were analyzed using a BD FACS Canto II flow cytometer (BD Biosciences) and FlowJo software (BD Biosciences).

[0162] Assessment of pulmonary edema and vascular leakage Lung wet / dry weight ratio was used as an index of fluid accumulation in the lungs. After mice were euthanized, lungs were surgically dissected, blotted dry, and immediately weighed (wet weight). Lung tissue was then dried in a 55°C oven for 72 hours and reweighed as dry weight. Wet to dry weight ratios were calculated for each animal to assess tissue edema (Tate et al., 2009; Tate et al., 2010). Protein concentration in cell-free BAL supernatants was measured by adding Bradford protein dye (Tate et al., 2009; Tate et al., 2010). A standard curve was generated using bovine serum albumin, and optical density (OD) was measured at 595 nm.

[0163] result As shown in Table 4, treatment with the cyclic peptide of SEQ ID NO: 1 (single dose of 10 mg / kg) routinely reduced polymorphonuclear cell (PMN) infiltration, viral titers and IL-6 levels in bronchial lavage fluid caused by viral infection. At a single dose of 10 mg / kg, peptides of SEQ ID NO: 1, 2, 9, 29, 37-39, 42, 56 and 59-61 were as effective as peptide of SEQ ID NO: 38 in reducing PMN infiltration in BAL fluid, whereas peptides of SEQ ID NO: 23, 40, 41, 50, 57 and 58 were relatively less effective than peptide of SEQ ID NO: 38 in reducing PMN infiltration in BAL fluid at a single dose of 10 mg / kg. Treatment with any one of the peptides of SEQ ID NO: 43, 44, 47-49 and 52 showed no change in PMN infiltration in BAL fluid at a single 10 mg / kg dose (activity scores: 0=inactive; 1=less active than peptide of SEQ ID NO: 38; 2=equal to or greater activity than peptide of SEQ ID NO: 38).

[0164] Compared to SEQ ID NO:38, treatment with any one of peptides SEQ ID NO:1, 9, 23, 29, 37, 38, 42, 50, 52, 56 and 59-61 (10 mg / kg single dose) was shown to be effective in reducing viral titers on day 3.

[0165] Although cytokine profiles varied in lung and serum samples, the data show that treatment with any one of peptides SEQ ID NO:1, 9, 23, 29, 37, 38, 42, 44, 47, 49, 50, 52, 56 and 59-61 was effective in reducing IL-6 levels in BAL fluid at levels comparable to peptide SEQ ID NO:38.

[0166] [Table 4]

[0167] Example 4: In vivo model of neuropathic pain This study was performed to evaluate the analgesic effect of the peptides described herein on neuropathic pain in vivo using the Chung rat nerve constriction model. Briefly, adult male Sprague-Dawley rats, 8-9 weeks old and weighing 220-250 g at the time of surgery, were purchased from Charles River UK Ltd.

[0168] Animals were housed in groups of four in an air-conditioned room with a 12-h light / dark cycle. Food and water were available ad libitum. Animals were allowed to acclimate to the experimental environment by remaining on an elevated metal mesh for at least 40 min for 3 days. Baseline paw withdrawal thresholds (PWTs) were assessed using a series of calibrated von Frey hairs for 3 consecutive days prior to surgery and reassessed 6–8 days after surgery and before drug administration on days 12–14 after surgery.

[0169] Each rat was anesthetized with 5% isoflurane mixed with oxygen (2 L per min) and then injected intramuscularly (i.m.) with ketamine 90 mg / kg + xylazine 10 mg / kg. The back was shaved and sterilized with povidone-iodine. The animals were placed in supine position and a paramedian incision was made in the skin overlying the L4-6 levels. The L5 spinal nerve was carefully isolated and tightly ligated with 6 / 0 silk suture. After complete hemostasis, the wound was closed in layers. A single dose of antibiotic (amoxipen, 15 mg / rat, i.p.) was routinely administered for postoperative infection prophylaxis. The animals were placed in a temperature-controlled recovery chamber until fully awake and then returned to their home cage.

[0170] Vehicle (1% DMSO in PBS) or peptide was administered intramuscularly (im) into the leg contralateral to the injury site by a second experimenter. Rats challenged with neuropathic pain conditions were randomly divided into five experimental groups: 1 mL / kg vehicle, 0.1, 0.5, 1 and 5 mg / kg peptide.

[0171] Each group had 8 animals. Animals were placed in individual Perspex boxes on an elevated metal mesh for at least 40 min prior to testing. Starting with the least stressful filament (approximately 1 g), each filament was applied perpendicular to the center of the ventral surface of the forepaw for 6 s until it was slightly bent. If the stimulus caused the animal to withdraw or lift the forepaw, the hair of the stress immediately below the one tested was used. If no response was observed, the hair of the stress immediately above was tested. The minimum amount of stress required to elicit a reliable response (positive in 3 out of 5 trials) was recorded as the value of PWT.

[0172] Drug testing was performed 12-14 days after surgery. PWT was assessed before and 1, 2, and 4 h after drug or vehicle administration. Animals were returned to their home cages and allowed to rest (approximately 30-60 min) between two adjacent test time points. Peptides were administered by a single intramuscular injection (IM) into the ipsilateral limb at doses of approximately 0.1 mg / kg body weight to approximately 5 mg / kg body weight.

[0173] result As shown in Table 5 below, peptides of SEQ ID NOs: 1, 2, 3, 10, 24 and 37, including the 6-mer peptide of SEQ ID NO: 39, reduced neuropathic pain in the Chung model following oral, subcutaneous and / or intramuscular administration (oral doses of 2-10 mg / kg, subcutaneous doses of 0.1-3 mg / kg, and intramuscular doses of 0.5-5 mg / kg).

[0174] [Table 5]

[0175] Example 5: In vivo model of systemic encephalomyocarditis virus (EMCV) infection In preliminary experiments using a mouse model of systemic encephalomyocarditis virus (EMCV) infection, a reduction in the number of neutrophils and inflammatory macrophages was observed in the peritoneal cavity after intraperitoneal administration of peptides of SEQ ID NO: 1, 37 and 38 (Table 6). This observation correlated with a reduction in circulating MCP-1, a cytokine that promotes the migration and activation of both of these immune cells.

[0176] [Table 6]

[0177] Example 6: In vivo model of neuropathic pain (II) The spinal nerve ligation (Chung) model was prepared as described in Example 4 above. Briefly, 64 adult male Sprague-Dawley rats, 8-9 weeks old and weighing 250-350 g at the time of surgery, were purchased from Charles River UK Ltd. The animals were housed in groups of four in an air-conditioned room with a 12-h light / dark cycle. Food and water were available ad libitum. The animals were allowed to acclimate to the experimental environment by being placed on an elevated metal mesh for at least 40 min for three days. Baseline paw withdrawal thresholds (PWTs) were determined using a series of calibrated von Frey hairs for three consecutive days prior to surgery and reassessed prior to drug administration on the 7th day after surgery and on the 12th to 14th day after surgery.

[0178] Each rat was anesthetized with 5% isoflurane mixed with oxygen (2 L per min) and then injected intramuscularly (i.m.) with ketamine 60 mg / kg + xylazine 10 mg / kg. The back was shaved and sterilized with povidone-iodine. The animals were placed in supine position and a paramedian incision was made in the skin overlying the L4-6 levels. The L5 spinal nerve was carefully isolated and tightly ligated with 6 / 0 silk suture. After complete hemostasis, the wound was closed in layers. A single dose of antibiotics (amoxipen, 15 mg / rat, i.p.) was routinely administered for postoperative infection prophylaxis. The animals were placed in a temperature-controlled recovery chamber until fully awake and then returned to their home cage.

[0179] Animals with a confirmed neuropathic pain state were randomly divided into four experimental groups: vehicle (first in 5% DMSO, then in 0.9% saline), 3 mg / kg LAT9997, 3 mg / kg LAT9997x1, and 3 mg / kg LAT1233x1. Each group contained six animals.

[0180] RSVEGS (SEQ ID NO:9; LAT9997), SVEGS (SEQ ID NO:62; LAT9997x1) and ALNSS (SEQ ID NO:63; LAT1233x1) were first dissolved in 5% DMSO and then in 0.9% saline, which also served as vehicle controls. All compounds were provided by GenScript to Lateral Pharma. All vehicles / compounds were administered intravenously at 1 mL / kg body weight.

[0181] Paw withdrawal threshold (PWT) Animals were placed in individual Perspex boxes on an elevated metal mesh for at least 40 min. Starting with the least stressful filament (1 gram (g)), each vFH filament was applied perpendicular to the center of the ventral surface of the forepaw for 6 s until it was slightly bent. If the stimulus caused the animal to withdraw or lift the forepaw, the filament of the next lower stress than the one tested was used. If no response was observed, the filament of the next higher stress was tested. The minimum amount of stress required to elicit a reliable response (positive in two out of three trials) was recorded as the value of PWT.

[0182] PWT was assessed once daily for 3 days before surgery (preD1, preD2, D0) and on day 7 after surgery to monitor the development of mechanical allodynia.

[0183] All drug tests were performed on days 13–17 after surgery. PWT was assessed before (BL), 1 h, and 2 h after drug or vehicle administration.

[0184] One-way analysis of variance (ANOVA) (IBM statistics SPSS, Version 27) was used for statistical analysis to compare PWT of different groups at the same time point. Fisher's least significant difference (LSD) post-hoc test was used to compare drug-treated and control groups, when appropriate. Paired Student's t-test (Microsoft Excel 365) was used to compare values ​​of the same group at different time points. To characterize drug-induced changes in PWT relative to vehicle, vehicle values ​​were subtracted from the appropriate drug values. The significance level was set at P<0.05.

[0185] result PWT in naive rats (before surgery) ranged from 10.0 to 15.0 g. The mean PWT of the ipsilateral (left) and contralateral (right) hindlimbs in the vehicle group on the day before surgery was 14.17 ± 0.83 g and 15.00 ± 0.00 g, respectively. The mean PWT of the LAT9997 group was 15.00 ± 0.00 g for both left and right hindlimbs, and the mean PWT of the LAT9997x1 and LAT1233x1 groups was 15.00 ± 0.00 g for both left and right hindlimbs. There were no statistically significant differences between groups (P > 0.05, one-way ANOVA).

[0186] On the 7th day after surgery, the ipsilateral PWT of the ligated nerve was significantly lower than the values ​​determined before surgery (6.00 ± 0.52 g in the vehicle group; 5.67 ± 0.33 g in the LAT9997 group; 6.33 ± 0.33 g in the LAT9997x1 group; and 5.33 ± 0.42 g in the LAT1233x1 group; P < 0.001 for all groups compared with preoperative values, paired Student's t-test). The contralateral PWT was not significantly affected by surgery (14.17 ± 0.83 g in the LAT1233x1 group; 15.00 ± 0.00 g in all other groups; P > 0.05 for all groups compared with preoperative values, paired Student's t-test).

[0187] Effect of vehicle (5% DMSO) on PWT Before administration of vehicle (5% DMSO) on the test day, the PWT of the (ipsilateral) hindpaw was significantly lower compared to the contralateral hindpaw: 3.33 ± 0.42 g ipsilaterally and 14.17 ± 0.83 g contralaterally (see Figures 3 and 4). After vehicle treatment, the ipsilateral PWT was not significantly affected from 1 to 4 h post-administration, being 3.67 ± 0.61 g, 3.67 ± 0.61 g, and 4.00 ± 0.89 g at the 1, 2, and 4 h time points, respectively (all P > 0.05, compared to pre-administration levels, paired Student's t-test, see Figure 3 and Table 7). On the contralateral side, the PWT was not affected (all 14.17 ± 0.83 g at all time points, see Figure 4 and Table 8).

[0188] Effect of LAT9997 on PWT At 3 mg / kg, LAT9997 induced a significant increase in the PWT of the ipsilateral hind paw in Chung model rats (see Figure 3 and Table 7). This effect was significant from 1 h post-dose: 3.33 ± 0.42 g before dosing vs. 7.83 ± 1.72 g at 1 h post-dose (P < 0.05, compared to pre-dose levels, paired Student's t-test). At 2 h post-dose, PWT further increased to 9.67 ± 1.73 g (P < 0.01, compared to pre-dose levels, paired Student's t-test). At 4 h post-dose, PWT slightly decreased to 8.17 ± 1.60 g (P < 0.05, compared to pre-dose levels, paired Student's t-test). PWT was significantly different from that recorded from the vehicle group at 2 and 4 h post-dose (both P < 0.05, one-way ANOVA).

[0189] Contralateral PWT did not change over the entire observation period (14.17±0.83 g pre-dose, 15.00±0.00 g at 1, 2, and 4 hours post-dose). Contralateral PWT was not significantly different from the vehicle group at any time point post-dose (P>0.05, one-way ANOVA, see Figure 4 and Table 8).

[0190] Effect of LAT1233x1 on PWT At 3 mg / kg, LAT1233x1 also induced a rapid and significant increase in the PWT of the ipsilateral hind paw of Chung model rats, starting 1 h after administration: 3.33 ± 0.42 g before administration, compared to 10.67 ± 1.67 g at 1 h after administration (P < 0.01, compared to pre-administration levels, paired Student's t-test). At 2 h after administration, the PWT increased slightly to 11.50 ± 1.80 g (P < 0.01, compared to pre-administration levels, paired Student's t-test). At 4 h after administration, the PWT decreased slightly to 10.17 ± 1.17 g (P < 0.01, compared to pre-administration levels, paired Student's t-test). At all time points after administration, the PWT was significantly different from that recorded from the vehicle group (all P < 0.01, one-way ANOVA; see Figure 3 and Table 7).

[0191] Contralateral PWT did not change significantly over the entire observation period (15.00±0.00 g before administration, 15.00±0.00 g, 14.17±0.83 g, and 15.00±0.00 g at 1, 2, and 4 hours after administration, respectively). Contralateral PWT was not significantly different from the vehicle group at any time point after administration (P>0.05, one-way ANOVA, see Figure 4 and Table 8).

[0192] [Table 7]

[0193] [Table 8]

[0194] [Table 9] TIFF2024525959000010.tif101159

Claims

1. A peptide capable of binding to the Lanthionine Synthetase C-like (LanCL) protein, wherein the peptide has an amino acid sequence of formula (I): X １ -X ２ -X ３ -X ４ -X ５ -X ６ (I) [wherein, (a) X １ is selected from the group consisting of lysine, arginine, and histidine, or X １ is absent, (b) X ２ is selected from the group consisting of alanine, valine, leucine, isoleucine, proline, phenylalanine, cysteine, tyrosine and serine, (c) X ３ is selected from the group consisting of glycine, alanine, valine, leucine and isoleucine, (d) X ４ is selected from the group consisting of serine, cysteine, threonine, asparagine, arginine, glutamine, tyrosine, aspartic acid, lysine, glutamic acid, proline, and histidine, or X ４ is absent, (e) X ５ is selected from the group consisting of serine, cysteine, threonine, asparagine, arginine, glutamine, tyrosine, lysine, histidine and glycine, or X ５ does not exist, and (f) X ６ is selected from the group consisting of serine, cysteine, threonine, asparagine, glutamine, tyrosine and histidine, or X ６ does not exist. ] including, the peptide is 3 to 12 amino acids in length, the amino acid sequence of the peptide does not include CRSRPVESSC, CRSVEGSCG or CRIIHNNNC, and the peptide is not a linear peptide containing the amino acid sequence EQLERALNSS.

2. The peptide according to claim 1, wherein the peptide includes the amino acid sequence SVEGS.

3. The peptide according to claim 1, wherein the peptide consists of the amino acid sequence SVEGS.

4. The peptide according to claim 1, wherein the peptide includes the amino acid sequence RSVEGS.

5. The peptide according to claim 1, wherein the peptide consists of the amino acid sequence RSVEGS.

6. The peptide according to claim 1, wherein the peptide includes an amino acid sequence selected from the group consisting of RAL, RALN, RALNS, RALNSS, RSV, RSVE, RSVEG, RSVEGS, RP, RPVE, RPVES, RPRESS, RII, RIIH, RIIHN and RIIHNN.

7. The peptide according to claim 1, wherein the peptide consists of an amino acid sequence selected from the group consisting of RAL, RALN, RALNS, RALNSS, RSV, RSVE, RSVEG, RSVEGS, RP, RPVE, RPVES, RPRESS, RII, RIIH, RIIHN and RIIHNN.

8. The peptide according to claim 1, wherein the peptide includes the amino acid sequence RALNSS.

9. The peptide according to claim 1, wherein the peptide consists of the amino acid sequence RALNSS. X １ The peptide according to claim 1, wherein X is absent.

10.

11. The peptide according to claim 10, wherein the peptide includes the amino acid sequence ALNSS.

12. The peptide according to claim 1, wherein one or more of the amino acids of formula (I) are D-amino acids.

13. A pharmaceutical composition comprising the peptide according to any one of claims 1 to 12.

14. The pharmaceutical composition according to claim 13, for treating a condition in a subject.

15. The pharmaceutical composition according to claim 14, wherein the condition is pain.

16. The pharmaceutical composition according to claim 15, wherein the condition is neuropathic pain.

17. The pharmaceutical composition according to claim 14, wherein the condition is an inflammatory airway disease.

18. The pharmaceutical composition according to claim 17, wherein the inflammatory airway disease is chronic obstructive pulmonary disease.

19. The pharmaceutical composition according to claim 14, wherein the condition is a respiratory infection.

20. The pharmaceutical composition according to claim 19, wherein the respiratory infection is a respiratory virus infection.

21. The pharmaceutical composition according to claim 20, wherein the virus is an influenza virus or a coronavirus.

22. Use of the peptide according to any one of claims 1 to 12 in the manufacture of a medicament for treating a condition in a subject.

23. The use according to claim 22, wherein the condition is selected from the group consisting of pain, inflammatory airway disease and respiratory infection.

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