Polypeptide analogues derived from annexin A1

Abstract

The present disclosure relates to polypeptides and polypeptide analogs derived from Annexin A1, and compositions comprising said polypeptides or polypeptide analogs for treating inflammatory and ischemic conditions.

Description

[Technical field]

[0001] The present disclosure relates to polypeptides and polypeptide analogs derived from Annexin A1, and compositions comprising said polypeptides or polypeptide analogs for treating inflammatory and ischemic conditions. [Background technology]

[0002] The annexin superfamily consists of 13 calcium phospholipid-binding proteins with significant biological and structural homology. Annexins are structurally divided into a highly conserved core domain and a variable N-terminal domain. Annexin A1 (ANXA1, 37 kDa-SEQ ID NO: 1) is an anti-inflammatory protein that inhibits blood-borne polymorphonuclear leukocyte (PMN) extravasation into surrounding tissues. The protein binds to the N-formyl peptide receptor (FPR)2 or FPR-L1 receptor, where it initiates a series of signaling events. Following inflammatory stimuli, migration of blood-borne polymorphonuclear leukocytes (PMN) into surrounding tissues occurs. PMN migration or extravasation is regulated by mediators such as adhesion molecules, cytokines, and proteases that control pro- and anti-inflammatory processes. PMNs have a high destructive capacity and are potentially self-damaging. Therefore, it is important to control PMN extravasation and inflammatory responses.

[0003] For therapeutic purposes as an anti-inflammatory agent, the complete annexin A1 protein has many disadvantages with respect to its functional fragments or modified forms. The large size of the protein makes it more difficult to deliver by techniques that are possible with smaller polypeptides (e.g., transdermal or transmucosal). When used to treat ocular inflammation, the smaller the size of the molecule, the better it is expected to penetrate the corneal epithelium. Also, susceptibility to proteolysis is a particular concern for all peptide pharmaceuticals, especially the larger ones, and especially when oral delivery (preferred by many patients) is intended.

[0004] Although several annexin A1 derivatives lacking key regions N-terminal to the polypeptide have been shown to lack significant activity in several assays of inflammation and mediator release, the full-length N-terminal N-acetyl annexin A1 (2-26) was deemed biologically active in several systems. Several peptides derived primarily from the unique N-terminal portion of the annexin A1 protein have been shown to have anti-inflammatory properties. One of the most extensively studied annexin A1 peptides is peptide Ac2-26, which mimics the 2nd to 26th amino acids of the 54 amino acid N-terminal region. Like the Ac1-188 fragment (and the native protein), it has N-terminal acetylation, which increases its stability and in some cases its half-life. Annexin A1 and its N-terminal peptide (Ac2-26) have been shown to exert most of their anti-inflammatory effects via the FPR2 / lipoxin A4 (FPR2 / Alx) receptor. In vivo, Ac2-26 peptide has been shown to exert anti-inflammatory effects in models of myocardial ischemia-reperfusion (I / R), mesenteric I / R, glycogen peritonitis and IL1 air pouch, and was reported to significantly reduce neutrophil recruitment to the site of injury / inflammation. The anti-inflammatory properties of this peptide are not limited to acute models of inflammation. In an arthritis model, intra-articular administration of Ac2-26 peptide was shown to reduce disease severity through reduction of neutrophil recruitment.

[0005] Shorter versions of the Ac2-26 peptide, such as peptides Ac2-12 and Ac2-6, have also been shown to elicit some anti-inflammatory effects in acute models of inflammation. Longer polypeptides with anti-inflammatory effects have been disclosed, such as those corresponding to amino acid residues 2-48 and 11-48 (WO2012 / 174397). Studies conducted by multiple laboratories have shown that a peptide derived from a region completely independent of the N-terminus of the annexin A1 protein, more precisely from amino acids 247-253 in the third repeat of the core region of the protein, called anti-flamin-2 (AF2), also has anti-inflammatory properties. Summary of the Invention

[0006] The present disclosure relates to annexin A1 polypeptide analogs, conjugates comprising annexin A1 polypeptides and branched amino acid probes (BAPs), and compositions comprising annexin A1 polypeptide analogs or conjugates thereof for treating ischemic and / or inflammatory conditions.

[0007] One aspect of the present disclosure relates to a polypeptide or polypeptide analog comprising at least the sequence AMVSEFLKQAWFIENEEQEYVQTVKS (SEQ ID NO: 2), or a functional variant thereof, wherein the polypeptide consists of a polypeptide selected from the group consisting of SEQ ID NO: 2 to SEQ ID NO: 24, or a functional variant thereof.

[0008] Another aspect of the disclosure relates to a polypeptide conjugate comprising the above-described polypeptide or polypeptide analog and one or more branched amino acid probes.

[0009] Another aspect of the disclosure relates to pharmaceutical compositions comprising the above-described polypeptides or polypeptide analogs.

[0010] A further aspect of the present disclosure relates to a polypeptide or polypeptide analog for use in the treatment of an ischemic and / or inflammatory condition. [Brief description of the drawings]

[0011] [Figure 1] In vitro phagocytosis by monocytes isolated from human blood. Results are shown as mean ± SEM from four experiments, each using monocytes from a different healthy volunteer. These data show that compound 1 (AnxA1 2-29(V24L)), compound 2 (AnxA1 2-34(V24L)), and compound 3 (AnxA1 2-39(V24L)) induce phagocytosis to a greater extent than AnxA1 2-50 (2.5-3 fold increase compared to vehicle). For details, see Example 1. [Diagram 2]In vitro chemotactic responses by polymorphonuclear leukocytes (PMN). These data show that compounds 1, 2, and 3 increase PMN chemotaxis compared to vehicle (1.6-3 fold increase). For details, see Example 2. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] The present disclosure relates to polypeptide analogs of annexin A1, conjugated forms of the polypeptide analogs, and compositions comprising the polypeptide analogs of annexin A1 or conjugated forms thereof. The disclosed polypeptide analogs, conjugates, and compositions are particularly effective for use in treating ischemic and inflammatory conditions.

[0013] Full-length annexin A1 (Homo sapiens) has the following sequence: >sp|P04083|ANXA1_HUMAN Annexin A1 OS=Homo sapiens GN=ANXA1 PE=1 SV=2 M AMVSEFLKQAWFIENEEQEYVQTVKS SKGGPGSAVSPYPTFNPSSDVAALHKAIMVKGVDEATIIDILTKRNNAQRQQIKAAYLQETGKPLDETLKKALTGHLEEVVLALLKTPAQFDADELRAAMKGLGTDEDTLIEILASRTNKEIRDINRVYREELKRDLAKDITSDTSGDFRNALLSLAKGDRSEDFGVNEDLADSDARALYEAGERRKGTDVNVFNTILTTRSYPQLRRVFQKYTKYSKHDMNKVLDLELKGDIEKCLTAIVKCATSKPAFFAEKLHQAMKGVGTRHKALIRIMVSRSEIDMNDIKAFYQKMYGISLCQAILDETKGDYEKILVALCGGN (SEQ ID NO: 1)

[0014] It is a first aspect to provide a polypeptide or polypeptide analogue comprising at least the sequence AMVSEFLKQAWFIENEEQEYVQTVKS (SEQ ID NO: 2), or a functional variant thereof, said polypeptide being selected from the group consisting of: AMVSEFLKQAWFIENEEQEYVQTVKS (SEQ ID NO:2) AMVSEFLKQAWFIENEEQEYVQTVKSS (SEQ ID NO: 3) AMVSEFLKQAWFIENEEQEYVQTVKSSK (SEQ ID NO: 4) AMVSEFLKQAWFIENEEQEYVQTVKSSKG (SEQ ID NO:5) AMVSEFLKQAWFIENEEQEYVQTVKSSKGG (SEQ ID NO: 6) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGP (SEQ ID NO: 7) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPG (SEQ ID NO: 8) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGS (SEQ ID NO: 9) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSA (SEQ ID NO: 10) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAV (SEQ ID NO: 11) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVS SEQ ID NO: 12) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSP (SEQ ID NO: 13) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPY (SEQ ID NO: 14) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYP (SEQ ID NO: 15) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPT (SEQ ID NO: 16) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTF (SEQ ID NO: 17) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTFN (SEQ ID NO: 18) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTFNP (SEQ ID NO: 19) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTFNPS (SEQ ID NO: 20) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTFNPSSD (SEQ ID NO: 21) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTFNPSSDVAALH (SEQ ID NO: 22) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTFNPSSDVAALHK (SEQ ID NO: 23) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTFNPSSDVAALHKA (SEQ ID NO: 24), or a functional variant thereof.

[0015] Also provided herein is a polypeptide selected from the group consisting of SEQ ID NO: 2 to SEQ ID NO: 24, or a functional variant thereof.

[0016] In a preferred embodiment, functional variants do not encompass functional fragments of the disclosed polypeptides or polypeptide analogs. Preferably, the polypeptides or polypeptide analogs of the invention have the length (i.e., the number of consecutive amino acids) specified in the SEQ ID NO.

[0017] Also disclosed herein is a functional variant of a polypeptide or polypeptide analog comprising at least the sequence AMVSEFLKQAWFIENEEQEYVQTVKS (SEQ ID NO:2), or a functional variant thereof, wherein the polypeptide consists of a functional variant of a polypeptide selected from the group consisting of any one of SEQ ID NO:2 to SEQ ID NO:24.

[0018] A polypeptide or polypeptide analogue defined herein (including functional variants thereof) in one embodiment activates and / or stimulates one or more of formyl peptide receptor 1 (FPR1), formyl peptide receptor 2 (FPR2), and formyl peptide receptor 3 (FPR3).

[0019] A functional polypeptide as defined herein, in one embodiment, is a ligand and / or agonist of one or more of formyl peptide receptor 1 (FPR1), formyl peptide receptor 2 (FPR2), and formyl peptide receptor 3 (FPR3).

[0020] The term "agonist" in this context refers to a polypeptide as defined herein that is capable of binding to, or in some embodiments at least to some extent, and / or activating a receptor, or in some embodiments at least to some extent, activating a receptor. Thus, for example, an FPR2 agonist is capable of binding to and / or activating FPR2.

[0021] An agonist can be an agonist of several different types of receptors and therefore can bind to and / or activate several different types of receptors. The agonist can also be a selective agonist that binds to and activates only one type of receptor. The term "antagonist" in this context refers to a substance that can inhibit the effect of a receptor agonist.

[0022] A full agonist binds to (has affinity for) a receptor, activates the receptor, and exhibits full efficacy at that receptor. A "partial agonist" in this context is a peptide that can bind to and activate a given receptor, but has only partial efficacy at the receptor relative to a full agonist. A partial agonist can act as an antagonist if it competes with a full agonist for receptor occupancy, resulting in a net decrease in receptor activation compared to the effect or activation observed with a full agonist alone.

[0023] A "selective agonist" in this context is a compound that is selective and therefore binds to and activates primarily one type of receptor. Thus, a selective FPR2 agonist is selective for FPR2.

[0024] A polypeptide or polypeptide analog according to the present disclosure, in one embodiment, is capable of binding to and activating one or several formyl peptide receptors to some degree and may have different binding affinities and / or different receptor activation efficacies for different receptors. Affinity refers to the number and size of intermolecular forces between a peptide ligand and its receptor, and the residence time of the ligand at its receptor binding site, and receptor activation efficacies refers to the ability of a peptide ligand to produce a biological response upon binding to a target receptor and the quantitative magnitude of this response. In some embodiments, such differences in affinity and receptor activation efficacies may be due to, for example, the EC for stimulating ligand binding in cells expressing one or more types of receptors described herein, or in tissues expressing various types of receptors. 50 The binding efficiency of the ligand is determined by receptor binding / activation studies routine in the art by generating binding efficiency values ​​and Emax values. High affinity means a lower concentration of ligand required to obtain 50% binding of the receptor compared to a ligand peptide with a lower affinity, and high receptor activation efficiency means a peptide with a lower affinity and / or receptor activation efficiency (high EC 50 A lower concentration of peptide required to obtain a 50% receptor activation response (lower EC50 value).

[0025] In one embodiment, the polypeptide may have different affinities and / or receptor activation efficacies for two or more receptors selected from FPR1, FPR2, and FPR3.

[0026] The receptor activation potency of a polypeptide agonist is also known as p(A 50 ) value.

[0027] In certain embodiments, a functional polypeptide has binding affinity and / or receptor efficacy for formyl peptide receptor 2 (FPR2), which may be tested using conventional methods or as outlined in the Examples.

[0028] In one particular embodiment, the polypeptide is capable of binding to and activating FPR2, hi a further embodiment, the peptide is a full agonist of FPR2.

[0029] In one embodiment, the polypeptide or polypeptide analog comprises at least the sequence AMVSEFLKQAWFIENEEQEYVQTVKS (SEQ ID NO: 2), or a functional variant thereof, wherein said polypeptide consists of a polypeptide selected from the group consisting of any one of SEQ ID NO: 2 to SEQ ID NO: 24, or a functional variant thereof, and is capable of one or more of the following: a) binding to one or more of the formyl peptide receptors, including FPR1, FPR2, and FPR3; and / or b) activating and / or stimulating one or more of the formyl peptide receptors, including FPR1, FPR2, and FPR3; and / or c) binding to and / or activating FPR2, and / or d) activating immune cells, and / or e) activating leukocytes, such as phagocytic leukocytes, e.g. monocytes; f) activating leukocytes, such as polymorphonuclear leukocytes (PMN), e.g. neutrophils; and / or g) activating neutrophils and / or monocytes; and / or h) activating leukocyte effector functions, such as one or more of the following: induction of neutrophil chemotaxis, recruitment of neutrophil complement receptor 3 (CR3), and activation of neutrophil NADPH-oxidase; and / or i) inducing phagocytosis in leukocytes, such as phagocytic leukocytes, e.g. monocytes, and / or j) Inducing chemotaxis in leukocytes such as polymorphonuclear leukocytes (PMN), e.g. neutrophils.

[0030] Variants of the polypeptides defined herein can in principle have one or more substitutions at one or more positions. Individual amino acid residues within the disclosed sequences can be substituted with any given proteinogenic or non-proteinogenic amino acid.

[0031] In one embodiment of the present disclosure, a functional variant of a polypeptide consisting of a polypeptide selected from the group consisting of any one of SEQ ID NOs: 2 to 24 has at least 75% sequence identity to any one of SEQ ID NOs: 2 to 24, for example, at least 80% sequence identity, for example, at least 85% sequence identity, for example, at least 90% sequence identity, for example, at least 95% sequence identity to any one of SEQ ID NOs: 2 to 24.

[0032] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 2 to SEQ ID NO: 24, having one or more amino acid substitutions. A single amino acid substitution means that an amino acid differs between the original and variant sequences at one position.

[0033] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 2 to SEQ ID NO: 24 having one amino acid substitution.

[0034] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 2 to SEQ ID NO: 24 having two amino acid substitutions.

[0035] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 2 to SEQ ID NO: 24 having three amino acid substitutions.

[0036] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 2 to SEQ ID NO: 24 having four amino acid substitutions.

[0037] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 2 to SEQ ID NO: 24 having five amino acid substitutions.

[0038] In one embodiment, the one or more amino acid substitutions are conservative amino acid substitutions. In one embodiment, the one or more amino acid substitutions are non-conservative amino acid substitutions.

[0039] The genetic code specifies 20 standard amino acids that are naturally incorporated into polypeptides (protein building blocks): Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Tyr, Thr, Trp, Val, and these are incorporated into proteins by a unique synthetic mechanism: Sec (selenocysteine, or U) and Pyl (pyrrolysine, O), all of which are L stereoisomers.

[0040] Besides the 22 standard or natural amino acids, there are many non-naturally occurring amino acids (non-proteinogenic or non-standard). They are not found in proteins or are not produced directly and isolated by standard cellular mechanisms. Non-standard amino acids are usually formed by modifications to standard amino acids, such as post-translational modifications. Examples of non-naturally occurring amino acid residues include Abu, Aib, Nle (norleucine), DOrn (D-ornithine, deguanylated arginine), Nal (β-2-naphthyl-alanine), D-Nal (β-2-naphthyl-D-alanine), DArg, DTrp, DPhe and DVal.

[0041] Any amino acid according to the present disclosure may be in the L or D configuration. If nothing is specified, reference to the L isomeric form is preferably meant.

[0042] The term "polypeptide" also includes post-translational modifications introduced by chemical or enzymatic catalysis, as known in the art. Such post-translational modifications can be introduced prior to cleavage, if necessary. Functional equivalents can also include chemical modifications such as ubiquitination, labeling (e.g., radionuclides, various enzymes, etc.), pegylation (derivatization with polyethylene glycol), or the insertion (or substitution by chemical synthesis) of amino acids that do not normally occur in human proteins (e.g., ornithine).

[0043] Polypeptides as defined herein having N-terminal alkylation and C-terminal esterification are also encompassed by the present disclosure.Functional equivalents also include glycosylation and covalent or aggregate conjugates formed with the same molecule, including dimers or unrelated chemical moieties.Such functional equivalents are prepared by linking functional groups to groups found within the fragments, including at either or both the N-terminus and C-terminus, by means known in the art.

[0044] In one embodiment, the alanine residue at position 10 of any one of SEQ ID NO:2 to SEQ ID NO:24 is substituted with any other standard or non-standard amino acid.

[0045] In one embodiment, the alanine residue at position 10 of any one of SEQ ID NOs: 2 to 24 is substituted with an amino acid residue independently selected from the group consisting of leucine, aspartic acid, methionine, glutamic acid, isoleucine, and arginine.

[0046] In one embodiment, the residue at position 10 of any one of SEQ ID NO:2 to SEQ ID NO:24 is not alanine, ie, it is any amino acid except alanine.

[0047] In one embodiment, the valine residue at position 21 of any one of SEQ ID NO:2 to SEQ ID NO:24 is substituted with any other standard or non-standard amino acid.

[0048] In one embodiment, the valine residue at position 21 of any one of SEQ ID NOs: 2 to 24 is substituted with an amino acid residue independently selected from the group consisting of leucine, aspartic acid, methionine, glutamic acid, isoleucine, and lysine.

[0049] In one embodiment, the residue at position 21 of any one of SEQ ID NO:2 to SEQ ID NO:24 is not a valine, ie, any amino acid except valine.

[0050] In one embodiment, the alanine residue at position 10 and the valine residue at position 21 of any one of SEQ ID NOs:2-24 are both substituted with an amino acid residue independently selected from the group consisting of leucine, aspartic acid, methionine, glutamic acid, isoleucine, arginine, and lysine; e.g., leucine, such as the group consisting of leucine, aspartic acid, and methionine. In one embodiment, the residue at position 10 and the residue at position 21 of any one of SEQ ID NOs:2-24 are identical.

[0051] In one embodiment, the valine residue at position 24 of any one of SEQ ID NO:2 to SEQ ID NO:24 is substituted with any other standard or non-standard amino acid.

[0052] In one embodiment, the residue at position 24 of any one of SEQ ID NO:2 to SEQ ID NO:24 is not a valine, ie, any amino acid except valine.

[0053] In one embodiment, the valine residue at position 24 of any one of SEQ ID NOs: 2 to 24 is substituted with an amino acid residue independently selected from the group consisting of glycine, alanine, serine, threonine, cysteine, leucine, isoleucine, methionine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, histidine, lysine, and arginine.

[0054] In a preferred embodiment, the valine residue at position 24 of any one of SEQ ID NO:2 to SEQ ID NO:24 is substituted with leucine. In some embodiments, the Val24Leu mutation increases the stability of the protease.

[0055] In one embodiment, the valine residue at position 35 of any one of SEQ ID NO:2 to SEQ ID NO:24 is substituted with any other standard or non-standard amino acid.

[0056] In one embodiment, the valine residue at position 35 of any one of SEQ ID NOs: 2 to 24 is substituted with an amino acid residue independently selected from the group consisting of glycine, alanine, serine, threonine, cysteine, leucine, isoleucine, methionine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, histidine, lysine, and arginine.

[0057] In one embodiment, the residue at position 35 of any one of SEQ ID NO:2 to SEQ ID NO:24 is a lysine.

[0058] In one embodiment, the residue at position 35 of any one of SEQ ID NO:2 to SEQ ID NO:24 is not a valine, ie, any amino acid except valine.

[0059] In one embodiment, the leucine residue at position 50 of any one of SEQ ID NO:2 to SEQ ID NO:24 is substituted with any other standard or non-standard amino acid.

[0060] In one embodiment, the leucine residue at position 50 of any one of SEQ ID NO: 23 to SEQ ID NO: 24 is substituted with an amino acid residue independently selected from the group consisting of glycine, alanine, serine, threonine, cysteine, valine, isoleucine, methionine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, histidine, lysine, and arginine.

[0061] In one embodiment, the residue at position 50 of any one of SEQ ID NO:23-24 is any amino acid except leucine.

[0062] In one embodiment, a polypeptide selected from the group consisting of: AMVSEFLKQX1WFIENEEQEYX2QTX3KS (SEQ ID NO:25), AMVSEFLKQX1WFIENEEQEYX2QTX3KSS (SEQ ID NO:26), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSK (SEQ ID NO:27), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKG (SEQ ID NO: 28), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGG (SEQ ID NO:29), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGP (SEQ ID NO: 30), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPG (SEQ ID NO:31), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPGS (SEQ ID NO:32), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPGSA (SEQ ID NO: 33), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPGSAX4 (SEQ ID NO: 34), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPGSAX4S (SEQ ID NO: 35), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPGSAX4SP (SEQ ID NO: 36), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPGSAX4SPY (SEQ ID NO: 37), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPGSAX4SPYP (SEQ ID NO: 38), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPGSAX4SPYPT (SEQ ID NO: 39), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPGSAX4SPYPTF (SEQ ID NO: 40), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPGSAX4SPYPTFN (SEQ ID NO: 41), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPGSAX4SPYPTFNP (SEQ ID NO: 42), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPGSAX4SPYPTFNPS (SEQ ID NO: 43), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPGSAX4SPYPTFNPSSD (SEQ ID NO: 44), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPGSAX4SPYPTFNPSSDVAAX5H (SEQ ID NO: 45), AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPGSAX4SPYPTFNPSSDVAAX5HK (SEQ ID NO: 46), and AMVSEFLKQX1WFIENEEQEYX2QTX3KSSKGGPGSAX4SPYPTFNPSSDVAAX5HKA (SEQ ID NO: 47), wherein X1 is selected from the group consisting of alanine, leucine, aspartic acid, methionine, glutamic acid, isoleucine, and arginine; wherein X2 is selected from the group consisting of valine, leucine, aspartic acid, methionine, glutamic acid, isoleucine, and lysine; wherein X3 is selected from the group consisting of valine, glycine, alanine, serine, threonine, cysteine, leucine, isoleucine, methionine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, histidine, lysine, and arginine, and is preferably leucine; wherein X4 is selected from the group consisting of valine, glycine, alanine, serine, threonine, cysteine, leucine, isoleucine, methionine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, histidine, lysine, and arginine, and is preferably lysine; and wherein X5 is selected from the group consisting of leucine, glycine, alanine, serine, threonine, cysteine, valine, isoleucine, methionine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, histidine, lysine, and arginine; or functional variants thereof having one or more amino acid substitutions, such as having 1, 2, 3, 4 or 5 amino acid substitutions at any position other than X1, X2, X3, X4, and X5.

[0063] In another embodiment, a polypeptide selected from the group consisting of: AMVSEFLKQAWFIENEEQEYVQTLKS (SEQ ID NO:48), AMVSEFLKQAWFIENEEQEYVQTLKSS (SEQ ID NO:49), AMVSEFLKQAWFIENEEQEYVQTLKSSK (SEQ ID NO:50), AMVSEFLKQAWFIENEEQEYVQTLKSSKG (SEQ ID NO:51), AMVSEFLKQAWFIENEEQEYVQTLKSSKGG (SEQ ID NO:52), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGP (SEQ ID NO:53), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPG (SEQ ID NO:54), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGS (SEQ ID NO:55), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSA (SEQ ID NO:56), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAV (SEQ ID NO:57), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVS (SEQ ID NO:58), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSP (SEQ ID NO:59), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPY (SEQ ID NO: 60), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYP (SEQ ID NO: 61), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPT (SEQ ID NO: 62), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPTF (SEQ ID NO: 63), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPTFN (SEQ ID NO: 64), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPTFNP (SEQ ID NO: 65), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPTFNPS (SEQ ID NO: 66), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPTFNPSSD (SEQ ID NO: 67), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPTFNPSSDVAALH (SEQ ID NO: 68), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPTFNPSSDVAALHK (SEQ ID NO: 69), and AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPTFNPSSDVAALHKA (SEQ ID NO: 70), or a functional variant thereof is provided.

[0064] In one embodiment of the present disclosure, a functional variant of a polypeptide consisting of a polypeptide selected from the group consisting of any one of SEQ ID NOs: 48 to 70 has at least 75% sequence identity to any one of SEQ ID NOs: 48 to 70, for example, at least 80% sequence identity, for example, at least 85% sequence identity, for example, at least 90% sequence identity, for example, at least 95% sequence identity to any one of SEQ ID NOs: 48 to 70.

[0065] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 48 to SEQ ID NO: 70, having one or more amino acid substitutions. A single amino acid substitution means that an amino acid differs between the original and variant sequences at one position.

[0066] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 48 to SEQ ID NO: 70 having one amino acid substitution.

[0067] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 48 to SEQ ID NO: 70 having two amino acid substitutions.

[0068] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 48 to SEQ ID NO: 70 having three amino acid substitutions.

[0069] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 48 to SEQ ID NO: 70 having four amino acid substitutions.

[0070] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 48 to SEQ ID NO: 70 having five amino acid substitutions.

[0071] In specific embodiments, the polypeptide or polypeptide analog comprises a peptide selected from the group consisting of: AMVSEFLKQAWFIENEEQEYVQTLKSSK (SEQ ID NO:50), AMVSEFLKQAWFIENEEQEYVQTLKSSKG (SEQ ID NO:51), AMVSEFLKQAWFIENEEQEYVQTLKSSKGG (SEQ ID NO:52), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGP (SEQ ID NO:53), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPG (SEQ ID NO:54), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGS (SEQ ID NO:55), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSA (SEQ ID NO:56), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAV (SEQ ID NO:57), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVS (SEQ ID NO:58), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSP (SEQ ID NO:59), and AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPY (SEQ ID NO: 60), or a functional variant thereof is provided.

[0072] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 50 to SEQ ID NO: 60, having one or more amino acid substitutions. A single amino acid substitution means that an amino acid differs between the original and variant sequences at one position.

[0073] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 50 to SEQ ID NO: 60 having one amino acid substitution.

[0074] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 50 to SEQ ID NO: 60 having two amino acid substitutions.

[0075] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 48 to SEQ ID NO: 70 having three amino acid substitutions.

[0076] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 50 to SEQ ID NO: 60 having four amino acid substitutions.

[0077] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 50 to SEQ ID NO: 60 having five amino acid substitutions.

[0078] In a preferred embodiment, the polypeptide or polypeptide analog is selected from the group: AMVSEFLKQAWFIENEEQEYVQTLKSSK (SEQ ID NO:50), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGS (SEQ ID NO:55), and AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPY (SEQ ID NO: 60), or a functional variant thereof.

[0079] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO: 50, SEQ ID NO: 55, and SEQ ID NO: 60, having one or more amino acid substitutions. A single amino acid substitution means that an amino acid differs between the original and variant sequences at one position.

[0080] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO:50, SEQ ID NO:55, and SEQ ID NO:60 having one amino acid substitution.

[0081] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO:50, SEQ ID NO:55, and SEQ ID NO:60 having two amino acid substitutions.

[0082] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO:50, SEQ ID NO:55, and SEQ ID NO:60 having three amino acid substitutions.

[0083] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO:50, SEQ ID NO:55, and SEQ ID NO:60 having four amino acid substitutions.

[0084] In one embodiment of the present disclosure, a functional variant of a polypeptide as defined herein is a polypeptide selected from the group consisting of any one of SEQ ID NO:50, SEQ ID NO:55, and SEQ ID NO:60 having five amino acid substitutions.

[0085] In one embodiment, the one or more amino acid substitutions are conservative amino acid substitutions. In one embodiment, the one or more amino acid substitutions are non-conservative amino acid substitutions.

[0086] In one embodiment of the present disclosure, the polypeptide or polypeptide analog, including any functional variant as defined herein, is acetylated.Any suitable residue in the polypeptide backbone or side chain may be acetylated.In one embodiment, the polypeptide is acetylated at N-terminus.

[0087] In one embodiment of the disclosure, a polypeptide or polypeptide analog, including any functional variant defined herein, is acetylated (COCH3 or Ac-) at the N-terminal amino acid residue (e.g., position 1, in one embodiment, alanine).

[0088] In one embodiment of the disclosure, a polypeptide or polypeptide analog, including any functional variant defined herein, is amidated (-NH2) at the C-terminus.

[0089] In one embodiment of the disclosure, the polypeptide, polypeptide analog or functional variant thereof comprises: a. binds to one or more of the formyl peptide receptors, including formyl peptide receptor 1 (FPR1), formyl peptide receptor 2 (FPR2), and formyl peptide receptor 3 (FPR3); and / or b. activating and / or stimulating one or more of the formyl peptide receptors, including formyl peptide receptor 1 (FPR1), formyl peptide receptor 2 (FPR2), and formyl peptide receptor 3 (FPR3); and / or c. is a ligand and / or agonist of one or more of the formyl peptide receptors, including formyl peptide receptor 1 (FPR1), formyl peptide receptor 2 (FPR2), and formyl peptide receptor 3 (FPR3), and / or d. binds to, activates, and / or is an agonist of FPR2; and / or e. Activating immune cells; and / or f. activating leukocytes, such as phagocytic leukocytes, e.g., monocytes, and / or g. Activating leukocytes, such as polymorphonuclear leukocytes (PMNs), e.g., neutrophils, and / or h. activating neutrophils and / or monocytes; and / or i. activating leukocyte effector functions, such as one or more of the following: induction of neutrophil chemotaxis, recruitment of neutrophil complement receptor 3 (CR3), and activation of neutrophil NADPH-oxidase; and / or j. Inducing phagocytosis in leukocytes, such as phagocytic leukocytes, e.g., monocytes; and / or k. Inducing chemotaxis in leukocytes such as polymorphonuclear leukocytes (PMNs), e.g., neutrophils.

[0090] Branched Amino Acid Probe (BAP) It is also an aspect of the present disclosure to provide a polypeptide or polypeptide analog of the present disclosure modified by the addition of one or more "branched amino acid probes". Branched amino acid probes (BAPs) are disclosed in WO2015 / 162485. In one embodiment, the polypeptide or polypeptide analog of the present disclosure is a conjugate comprising the above-mentioned polypeptide and one or more branched amino acid probes.

[0091] In some embodiments, the polypeptide or polypeptide analog conjugates provided herein have certain improved properties compared to the corresponding native or unconjugated polypeptide. In one embodiment, the BAP conjugates provided herein have increased binding affinity and / or activation of one or more relevant receptors, such as FPR. In another embodiment, the BAP conjugates provided herein are more stable, such as less susceptible to proteases. Furthermore, in one embodiment, the BAP conjugates have higher solubility.

[0092] In one aspect, a polypeptide conjugate is provided comprising a polypeptide or polypeptide analog comprising at least the sequence AMVSEFLKQAWFIENEEQEYVQTVKS (SEQ ID NO:2), or a functional variant thereof, wherein said polypeptide comprises a polypeptide selected from the group consisting of SEQ ID NO:2 to SEQ ID NO:24, or a functional variant thereof, and one or more branched amino acid probes consisting of 2 to 9 amino acid residues, wherein the branch point of the branched amino acid probe is formed by an amide bond between the ε-amino group of one amino acid residue and the carboxyl group of another amino acid residue, all other bonds are regular peptide bonds, and wherein one or more alpha amino groups of the branched amino acid probe are optionally acetylated.

[0093] In one aspect, a polypeptide conjugate is provided comprising a polypeptide or polypeptide analog comprising at least the sequence AMVSEFLKQAWFIENEEQEYVQTVKS (SEQ ID NO: 2), or a functional variant thereof, wherein the polypeptide comprises a polypeptide selected from the group consisting of SEQ ID NO: 2 to SEQ ID NO: 24, or a functional variant thereof, and one or more branched amino acid probes; wherein the branched amino acid probe comprises a first aminoalkyl amino acid residue; the first aminoalkyl amino acid residue (AAA) is optionally covalently linked to a second aminoalkyl amino acid residue, or to a second and a third aminoalkyl amino acid residue, so as to form a linear chain of two or three aminoalkyl amino acid residues; the side chain of one or more of the first, second and / or third aminoalkyl amino acid residues is modified by attachment to the side chain amino group of a molecule independently selected from the group consisting of AAAq-AAA, (aa3)p-AAAq, AAAq-(aa3)p, [(aa3)-AAA]p and [AAA-(aa3)]p, wherein q is a number selected from 0, 1, 2, and 3; p is a number selected from 1, 2, and 3; AAA is an aminoalkyl amino acid residue; and (aa3) is an amino acid residue independently selected from Arg, His, Gly, and Ala; the first aminoalkyl amino acid residue is covalently attached to the N-terminus of the polypeptide or polypeptide analog, covalently attached to the C-terminus of the polypeptide or polypeptide analog, and / or covalently attached to a side chain amino group of an aminoalkyl amino acid residue within the polypeptide or polypeptide analog; However, the branched amino acid probe must consist of 2 to 9 amino acid residues.

[0094] It is a further aspect to provide a polypeptide conjugate comprising a polypeptide selected from the group consisting of SEQ ID NO:2 to SEQ ID NO:24, SEQ ID NO:25 to SEQ ID NO:47, and SEQ ID NO:48 to SEQ ID NO:70, or a functional variant thereof, and one or more branched amino acid probes consisting of 2 to 9 amino acid residues, wherein the branch point of the branched amino acid probe is formed by an amide bond between the ε-amino group of one amino acid residue and the carboxyl group of another amino acid residue, all other bonds are regular peptide bonds, and one or more alpha amino groups of the branched amino acid probe are optionally acetylated.

[0095] In one aspect, there is provided a polypeptide conjugate comprising a polypeptide selected from the group consisting of SEQ ID NO:2 to SEQ ID NO:24, SEQ ID NO:25 to SEQ ID NO:47, and SEQ ID NO:48 to SEQ ID NO:70, or a functional variant thereof, and one or more branched amino acid probes, wherein the branched amino acid probe comprises a first aminoalkyl amino acid residue, the first aminoalkyl amino acid residue (AAA) is optionally covalently linked to a second aminoalkyl amino acid residue, or to a second and a third aminoalkyl amino acid residue, so as to form a linear chain of two or three aminoalkyl amino acid residues; the side chain of one or more of the first, second and / or third aminoalkyl amino acid residues is modified by attachment to the side chain amino group of a molecule independently selected from the group consisting of AAAq-AAA, (aa3)p-AAAq, AAAq-(aa3)p, [(aa3)-AAA]p and [AAA-(aa3)]p, wherein q is a number selected from 0, 1, 2, and 3; p is a number selected from 1, 2, and 3; AAA is an aminoalkyl amino acid residue; and (aa3) is an amino acid residue independently selected from Arg, His, Gly, and Ala; the first aminoalkyl amino acid residue is covalently attached to the N-terminus of the polypeptide or polypeptide analog, covalently attached to the C-terminus of the polypeptide or polypeptide analog, and / or covalently attached to a side chain amino group of an aminoalkyl amino acid residue within the polypeptide or polypeptide analog; However, the branched amino acid probe must consist of 2 to 9 amino acid residues.

[0096] In one embodiment, the N-terminal amino acid residue of the molecule is acetylated at the alpha amino group.

[0097] In one embodiment, the first aminoalkyl amino acid residue is linked by a peptide bond (amide) formed by reaction of a carboxylic acid or derivative thereof of the first aminoalkyl amino acid with the alpha amino group of the N-terminal amino acid residue of the polypeptide, by a peptide bond to the C-terminal amino acid residue of the polypeptide formed by reacting the alpha amino group of the aminoalkyl amino acid residue with a carboxylic acid or derivative thereof of the C-terminal amino acid residue, and / or is linked to an aminoalkyl amino acid residue in the polypeptide by an amide formed by reaction of a carboxylic acid or derivative thereof of the first aminoalkyl amino acid residue with the alkyl amino group of the aminoalkyl amino residue.

[0098] As defined herein, in one embodiment, an amino acid residue and / or peptide covalently attached to a further amino acid residue refers to the presence of a peptide bond, while in another embodiment, an amino acid residue covalently attached to the side chain amino group of an aminoalkyl amino acid residue in said polypeptide refers to the presence of an amide bond.

[0099] A peptide bond (amide bond) is a covalent chemical bond that forms between two molecules when a carboxyl group of one molecule reacts with an amino group of the other, causing the release of a molecule of HO. The process usually occurs between amino acids.

[0100] When the branched amino acid probe is covalently attached to the N-terminus of the polypeptide, the N-terminal amino alkyl amino acid residue of the backbone of the branched amino acid probe is preferably acetylated.

[0101] When the branched amino acid probe is covalently attached to the side chain amino group of an aminoalkyl amino acid residue in the polypeptide, the N-terminal aminoalkyl amino acid residue of the backbone of the branched amino acid probe is preferably acetylated.

[0102] When the branched amino acid probe is covalently attached to the C-terminus of the polypeptide, the C-terminal amino alkyl amino acid residue of the backbone of the branched amino acid probe is preferably a carboxylic acid, aldehyde, ester, or amide, such as a primary amide, and is most preferably amidated.

[0103] The aminoalkyl amino acid residue (or AAA) and the amino acid residue (aa3) according to the present disclosure may each be the same (identical) or different (non-identical).

[0104] Aminoalkylamino acid residues According to the present disclosure, each branched amino acid probe comprises at least one, such as two, such as three, aminoalkyl amino acid residue(s).

[0105] According to the present disclosure, an "aminoalkyl amino acid residue" (or AAA) is an amino acid having conventional amine (-NH) and carboxylic acid (-COOH) functional groups and a side chain covalently attached to the first (α-) carbon atom, the side chain containing an aminoalkyl group (-C n H 2n NH2).

[0106] Thus, an aminoalkyl amino acid residue (or AAA) is an aminoalkyl group (-C n H 2nNH2), which in one embodiment represents a side chain aminoalkyl group.

[0107] In one embodiment, the side chain alkyl group is methyl (CH3-), ethyl (C2H5-), propyl (C3H7-), butyl (C4H9-), pentyl (C5H 11 -), hexyl (C6H 13 -), heptyl (C7H15-), octyl (C8H 17 -), nonyl (C9H 19 -), Decyl (C 10 H 21 -), undecyl (C 11 H 23 -), and dodecyl (C 12 H 25 When an alkyl residue having a specific number of carbons is named, all geometric isomers having that number of carbons are intended to be encompassed, thus, for example, "butyl" is intended to include n-butyl, sec-butyl, isobutyl, and t-butyl.

[0108] In one embodiment, the side chain amino group (NH2) of the aminoalkyl amino acid residue is an amine of methylamine, an amine of ethylamine, an amine of propylamine, an amine of n-butylamine, an amine of pentylamine, an amine of n-hexylamine, an amine of heptylamine, an amine of octylamine, an amine of nonylamine, an amine of decylamine, an amine of undecylamine, or an amine of dodecylamine.

[0109] In one embodiment, the side chain aminoalkyl group is methylamine (-CH2NH2), ethylamine (-C2H4NH2), propylamine (-C3H6NH2), n-butylamine (-C4H8NH2), pentylamine (-C5H 10 NH2), n-Hexylamine (-C6H 12 NH2), heptylamine (-C7H 14 NH2), Octylamine (-C8H 16 NH2), nonylamine (-C9H 18NH2), decylamine (-C 10 H 20 NH2), undecylamine (-C 11 H 22 NH2), and dodecylamine (-C 12 H 24 NH2).

[0110] In one embodiment of the disclosure, the side chain amino group (NH2) of the first, second and / or third aminoalkyl amino acid residues contained in each BAP is each modified by attaching a molecule thereto.

[0111] In one embodiment, the side chain amino group of the aminoalkyl amino acid residue in each BAP is β (beta) amino group (one methylene in the side chain, methylamine), γ (gamma) amino group (two methylenes in the side chain, ethylamine), δ (delta) amino group (three methylenes in the side chain, propylamine), = ornithine ε (epsilon) amino group (four methylenes in the side chain, n-butylamine), = lysine ζ (zeta) amino group (five methylenes in the side chain, pentylamine), η (eta) amino group (six methylenes in the side chain, n-hexylamine), θ (theta) amino group (seven methylenes in the side chain, heptylamine), iotaamino group (8 methylenes in the side chain, octylamine), κ (kappa) amino group (nine methylenes in the side chain, nonylamine), Lambda amino group (10 methylenes in the side chain, decylamine), μ (mu) amino group (11 methylenes in the side chain, undecylamine), and It is selected from the group consisting of ν (nu) amino groups (12 methylenes in the side chain, dodecylamine).

[0112] For example, the ε-amino group is covalently bonded to the fifth carbon starting from (and including) the α-carbon, which is covalently bonded to a carboxyl (C═OOH) group.

[0113] An aminoalkylamino acid residue in which the side chain is an n-butylamine and the side chain amino group is an ε (epsilon) amino group is lysine (Lys, K).

[0114] Similarly, the δ-amino group is covalently attached to the fourth carbon starting from the α-carbon.

[0115] An aminoalkylamino acid residue in which the side chain is a propylamine and the side chain amino group is a δ (delta) amino group is ornithine (Orn).

[0116] Ornithine is formed in cells by the deguanidination of arginine. Although it is not used in protein synthesis in vivo, it is a participant in several enzymatic pathways and may play a role in nitrogen balance in vivo, as it can be enzymatically guanidinated to form arginine.

[0117] Any amino acid according to the present disclosure may be in the L or D configuration. If nothing is specified, reference to the L isomeric form is preferably meant.

[0118] Thus, in one embodiment, the aminoalkyl amino acid residues are individually in the L or D configuration. In one embodiment, the aminoalkyl amino acid residues are in the L configuration.

[0119] In one embodiment, the aminoalkyl amino acid residues contained in the branched amino acid probe are individually selected from the group consisting of lysine and ornithine.

[0120] In one embodiment, the aminoalkyl amino acid residue is selected from the group consisting of lysine and D-lysine, hi certain embodiments, the aminoalkyl amino acid residue is a lysine residue.

[0121] In one embodiment, the aminoalkyl amino acid residue is selected from the group consisting of ornithine and D-ornithine.

[0122] In one embodiment of the disclosure, there is provided a polypeptide or polypeptide analogue as defined herein, and one or more branched amino acid probes, the branched amino acid probe comprises a first amino acid residue selected from lysine and ornithine; the first amino acid residue is optionally covalently linked to a second amino acid residue or a second and a third amino acid residue selected from lysine or ornithine to form a linear chain of two or three lysine or ornithine residues; The side chain of one or more of the first, second and / or third lysine or ornithine residues is at the δ-amino group (ornithine) or at the ε-amino group (lysine); Lys q -Lys, (aa3) p -Lys q , Lys q -(aa3) p , [(aa3)-Lys] p , [Lys-(aa3)] p , Orn q -Orn, (aa3) p -Orn q , Orn q -(aa3) p , [(aa3)-Orn] p and [Orn-(aa3)] p , Orn p -Lys p , Lys p -Orn p , [Orn-Lys] p and [Lys-Orn] p and wherein the modified molecule is independently selected from the group consisting of wherein q is a number selected from 0, 1, 2, and 3; p is a number selected from 1, 2, and 3; and (aa3) is an amino acid residue independently selected from Arg, His, Gly, and Ala; the first lysine or ornithine residue is covalently linked to the N-terminus of the polypeptide or polypeptide analog, covalently linked to the C-terminus of the polypeptide or polypeptide analog, and / or covalently linked to an ε-amino group of a lysine residue or a δ-amino group of an ornithine residue within the polypeptide or polypeptide analog; However, the branched amino acid probe must consist of 2 to 9 amino acid residues.

[0123] In one embodiment there is provided a polypeptide or polypeptide analogue as defined herein and one or more branched amino acid probes, the branched amino acid probe comprises a first lysine residue, the first lysine residue is optionally covalently linked to a second lysine residue or to a second and a third lysine residue to form a linear chain of two or three lysine residues; The side chain of one or more of the first, second, and / or third lysine residues is at the ε-amino group of the lysine. q -Lys, (aa3) p -Lys q , Lys q -(aa3) p , [(aa3)-Lys] p , [Lys-(aa3)] p wherein q is a number selected from 0, 1, 2, and 3; p is a number selected from 1, 2, and 3; and (aa3) is an amino acid residue independently selected from Arg, His, Gly, and Ala; the first lysine residue is covalently linked to the N-terminus of the polypeptide or polypeptide analog, covalently linked to the C-terminus of the polypeptide or polypeptide analog, and / or covalently linked to an ε-amino group of a lysine or a δ-amino group of an ornithine residue within the polypeptide or polypeptide analog; However, the branched amino acid probe must consist of 2 to 9 amino acid residues.

[0124] Probe branch In one embodiment, a branched amino acid probe (BAP) according to the present disclosure consists of 2 to 9 amino acid residues.

[0125] In one embodiment of the present disclosure, each of the one or more branched amino acid probes consists of 2 to 3 amino acid residues, such as 3 to 4 amino acid residues, for example, 4 to 5 amino acid residues, such as 5 to 6 amino acid residues, for example, 6 to 7 amino acid residues, such as 7 to 8 amino acid residues, for example, 8 to 9 amino acid residues.

[0126] In one embodiment of the disclosure, each of the one or more branched amino acid probes consists of 2 amino acid residues, such as 3 amino acid residues, for example, 4 amino acid residues, such as 5 amino acid residues, for example, 6 amino acid residues, such as 7 amino acid residues, for example, 8 amino acid residues, such as 9 amino acid residues. In a particular embodiment of the disclosure, each of the one or more branched amino acid probes consists of 3 amino acid residues.

[0127] In one embodiment of the present disclosure, the branched amino acid probe comprises a first aminoalkyl amino acid residue (also designated AAA1), which is attached to a polypeptide or polypeptide analogue as defined herein to provide a polypeptide conjugate according to the present disclosure.

[0128] In one embodiment of the disclosure, the first aminoalkyl amino acid of the one or more branched amino acid probe(s) is covalently attached to the N-terminus of a polypeptide or polypeptide analog of the disclosure, covalently attached to the C-terminus of the polypeptide, and / or to the side chain amino group of an aminoalkyl amino acid residue within the polypeptide.

[0129] In one embodiment of the present disclosure, the branched amino acid probe comprises a first aminoalkyl amino acid residue, in one embodiment the side chain of which is modified by attachment of a molecule as defined herein to the side chain amino group.

[0130] In one embodiment of the present disclosure, the first aminoalkyl amino acid of the branched amino acid probe is acetylated at the alpha amino group. In one embodiment of the present disclosure, the N-terminus of the first aminoalkyl amino acid residue of the branched amino acid probe is acetylated.

[0131] In one embodiment of the present disclosure, when a branched amino acid probe comprising said first aminoalkyl amino acid residue is covalently attached to the N-terminus of a polypeptide defined herein, or when a branched amino acid probe comprising said first aminoalkyl amino acid residue is covalently attached to the side chain amino group of an aminoalkyl amino acid residue within said polypeptide, the N-terminus of the first aminoalkyl amino acid residue of the branched amino acid probe is acetylated.

[0132] In one embodiment of the present disclosure, the C-terminus of the first aminoalkyl amino acid residue of the branched amino acid probe is a carboxylic acid, an aldehyde, an ester, or an amide, such as a primary amide (CONH2). In a preferred embodiment of the present disclosure, the C-terminus of the first aminoalkyl amino acid residue is amidated.

[0133] In one embodiment of the present disclosure, when a branched amino acid probe comprising said first aminoalkyl amino acid residue is covalently attached to the C-terminus of a polypeptide defined herein, the C-terminus of the first aminoalkyl amino acid residue of the branched amino acid probe is an amide.

[0134] In one embodiment of the disclosure, the first aminoalkyl amino acid residue is covalently linked to a second aminoalkyl amino acid residue to form a linear chain of two aminoalkyl amino acid residues. In one embodiment of the disclosure, the alpha-amino group of the second aminoalkyl amino acid residue of the branched amino acid probe is acetylated. In one embodiment, the N-terminus of the second aminoalkyl amino acid residue of the branched amino acid probe is acetylated.

[0135] In one embodiment of the present disclosure, the C-terminus of the second aminoalkyl amino acid residue of the branched amino acid probe is a carboxylic acid, an aldehyde, an ester, or an amide, such as a primary amide (CONH2). In a preferred embodiment of the present disclosure, the C-terminus of the second aminoalkyl amino acid residue is amidated.

[0136] In one embodiment of the disclosure, the first aminoalkyl amino acid residue is covalently linked to a second and a third (covalently linked) aminoalkyl amino acid residue to form a linear chain of three aminoalkyl amino acid residues.

[0137] In one embodiment of the disclosure, the alpha-amino group of the third aminoalkyl amino acid residue of the branched amino acid probe is acetylated. In one embodiment of the disclosure, the N-terminus of the third aminoalkyl amino acid residue of the branched amino acid probe is acetylated.

[0138] In one embodiment of the present disclosure, the C-terminus of the third aminoalkyl amino acid residue of the branched amino acid probe is a carboxylic acid, an aldehyde, an ester, or an amide, such as a primary amide (CONH2). In a preferred embodiment of the present disclosure, the C-terminus of the third aminoalkyl amino acid residue is amidated.

[0139] In one embodiment of the disclosure, a first aminoalkyl amino acid residue has both a second and a third aminoalkyl amino acid residue covalently attached to its amine group. In one embodiment of the disclosure, a first aminoalkyl amino acid residue has both a second and a third aminoalkyl amino acid residue covalently attached to its carboxylic acid group. In one embodiment, a first aminoalkyl amino acid residue has a second aminoalkyl amino acid residue attached to its amine group and a third aminoalkyl amino acid residue attached to its carboxylic acid group.

[0140] The second and third aminoalkyl amino acid residues may be designated AAA2 and AAA3, respectively.

[0141] In one embodiment of the disclosure, each of the first, second and / or third aminoalkyl amino acid residues is methylamine (-CH2NH2), ethylamine (-C2H4NH2), propylamine (-C3H6NH2), n-butylamine (-C4H8NH2), pentylamine (-C5H 10 NH2), n-Hexylamine (-C6H 12 NH2), heptylamine (-C7H 14 NH2), Octylamine (-C8H 16 NH2), nonylamine (-C9H 18 NH2), decylamine (-C 10 H 20 NH2), undecylamine (-C 11 H 22 NH2), and dodecylamine (-C 12 H 24 NH2).

[0142] In one embodiment of the present disclosure, each of the first, second and / or third aminoalkyl amino acid residues of the branched amino acid probe is individually selected from the group consisting of lysine, D-lysine, ornithine and D-ornithine.

[0143] In one embodiment of the present disclosure, each of the first, second and third aminoalkyl amino acid residues of the branched amino acid probe is a lysine residue (including L-lysine and D-lysine).

[0144] In one embodiment of the present disclosure, the first, second or third aminoalkyl amino acid residue of the branched amino acid probe is acetylated at the alpha amino group (Ac-AAA)(COCH3).

[0145] In one embodiment of the present disclosure, the first, first and second, and the first, second and third aminoalkyl amino acid residues of the branched amino acid probe are each an aminoalkyl amino acid backbone (AAA1, AAA 1-2 , AAA 1-3 ) is called.

[0146] In one embodiment of the present disclosure, the first, first and second, and first, second and third aminoalkyl amino acid residues are each a lysine residue. In one embodiment, the first, first and second, and first, second and third lysine residues of the branched amino acid probe are each a lysine backbone (Lys1, Lys2, Lys3, Lys4, Lys5, Lys6, Lys7, Lys8, Lys9, Lys10, Lys11, Lys12, Lys13, Lys14, Lys15, Lys16, Lys17, Lys18, Lys19, Lys20, Lys21, Lys22, Lys23, Lys24, Lys25, Lys26, Lys27, Lys28, Lys29, Lys210, Lys211, Lys22, Lys23, Lys24, Lys25, Lys26, Lys27 1-2 , Lys 1-3 ) is called.

[0147] In one embodiment of the present disclosure, the first lysine residue, or the second lysine residue, or the third lysine residue of the lysine backbone of the branched amino acid probe is acetylated at the alpha-amino group (Ac-Lys).

[0148] In one embodiment of the disclosure, the side chain of one of the first, second and / or third aminoalkyl amino acid residues is modified by attaching a molecule as defined herein to the side chain amino group.

[0149] In one embodiment of the present disclosure, the branched amino acid probe comprises a first aminoalkyl amino acid residue, the side chain of which is modified by attaching a molecule as defined herein to the side chain amino group.

[0150] In one embodiment of the present disclosure, the branched amino acid probe comprises a first and a second aminoalkyl amino acid residue, the side chain of the first aminoalkyl amino acid residue being modified by attachment of a molecule as defined herein to the side chain amino group.

[0151] In one embodiment of the present disclosure, the branched amino acid probe comprises a first and a second aminoalkyl amino acid residue, the side chain of the second aminoalkyl amino acid residue being modified by attachment of a molecule as defined herein to the side chain amino group.

[0152] In one embodiment of the present disclosure, the branched amino acid probe comprises a first and a second aminoalkyl amino acid residue, the side chains of which are modified by attachment of a molecule as defined herein to the side chain amino group.

[0153] In one embodiment of the disclosure, the side chains of two of the first, second and / or third aminoalkyl amino acid residues are modified by attaching a molecule as defined herein to the side chain amino group.

[0154] In one embodiment of the disclosure, all three side chains of the first, second and third aminoalkyl amino acid residues are modified by attaching a molecule as defined herein to the side chain amino groups.

[0155] In one embodiment of the disclosure, the side chain of i) the first aminoalkyl amino acid residue, ii) the second aminoalkyl amino acid residue, iii) the third aminoalkyl amino acid residue, iv) the first and second aminoalkyl amino acid residues, v) the first and third aminoalkyl amino acid residues, vi) the second and third aminoalkyl amino acid residues, or vii) the first, second and third aminoalkyl amino acid residues is modified by attaching a molecule as defined herein to the side chain amino group.

[0156] In one embodiment of the disclosure, the first lysine residue, or the second lysine residue, or the third lysine residue, or the first and second lysine residues, or the first and third lysine residues, or the second and third lysine residues, or the first, second and third lysine residues of the lysine backbone of the branched amino acid is modified by attaching a molecule to the ε-amino group.

[0157] In one embodiment of the disclosure, one or more side chains of each of the first, second and / or third aminoalkyl amino acid residues are AAA q -AAA, (aa3) p -AAA q , AAA q -(aa3) p , [(aa3)-AAA] p and [AAA-(aa3)] p where q is a number selected from 0, 1, 2, and 3, p is a number selected from 1, 2, and 3, AAA is an amino alkyl amino acid residue, and (aa3) is an amino acid residue independently selected from Arg, His, Gly, and Ala. In one embodiment, the N-terminal AAA or (aa)3 of the molecule is acetylated at the alpha amino group.

[0158] In one embodiment of the disclosure, one or more side chains of each of the first, second and / or third aminoalkyl amino acid residues are Lys q -Lys, (aa3) p -Lys q , Lys q -(aa3) p , [(aa3)-Lys] p , [Lys-(aa3)] p , Orn q -Orn, (aa3) p -Orn q , Orn q -(aa3) p , [(aa3)-Orn] p and [Orn-(aa3)]p , Orn p -Lys p , Lys p -Orn p , [Orn-Lys] p and [Lys-Orn] p wherein q is a number selected from 0, 1, 2, and 3, p is a number selected from 1, 2, and 3, and (aa3) is an amino acid residue independently selected from Arg, His, Gly, and Ala. In one embodiment, the N-terminal Lys, Orn, or (aa)3 of the molecule is acetylated at the alpha amino group.

[0159] In one embodiment of the disclosure, one or more side chains of each of the first, second and / or third aminoalkyl amino acid residues are selected from the group consisting of Lys, q -Lys, (aa3) p -Lys q , Lys q -(aa3) p , [(aa3)-Lys] p and [Lys-(aa3)] p where q is a number selected from 0, 1, 2, and 3, p is a number selected from 1, 2, and 3, Lys is a lysine residue selected from L-Lys and D-Lys, and (aa3) is an amino acid residue independently selected from Arg, His, Gly, and Ala. In one embodiment, the N-terminal Lys or (aa)3 of the molecule is acetylated at the alpha amino group.

[0160] In one embodiment of the disclosure, one or more side chains of each of the first, second and / or third lysine residues of the lysine backbone are Lys q -Lys, (aa3) p -Lys q , Lys q -(aa3) p , [(aa3)-Lys] pand [Lys-(aa3)] p where q is a number selected from 0, 1, 2, and 3, p is a number selected from 1, 2, and 3, Lys is a lysine residue selected from L-Lys and D-Lys, and (aa3) is an amino acid residue independently selected from Arg, His, Gly, and Ala. In one embodiment, the N-terminal Lys or (aa)3 of the molecule is acetylated at the alpha amino group.

[0161] In one embodiment of the disclosure, one or more side chains of each of the first, second, and / or third lysine residues of the lysine backbone are Lys q -Lys, where q is a number selected from 0, 1, 2, and 3. In one embodiment, the N-terminal Lys of the molecule is acetylated at the alpha amino group.

[0162] In one embodiment of the present disclosure, the molecules covalently attached to the ε-amino group of one or more lysine residues of the lysine backbone of the branched amino acid probe are independently Lys q -Lys, (aa3) p -Lys q , Lys q -(aa3) p , [(aa3)-Lys] p and [Lys-(aa3)] p wherein q is a number selected from 0, 1, 2, and 3, p is a number selected from 1, 2, and 3, and (aa3) is an amino acid residue independently selected from Arg, His, Gly, and Ala. In one embodiment, the N-terminal Lys or (aa)3 of the molecule is acetylated at the alpha amino group.

[0163] Thus, in one embodiment of the present disclosure, the first lysine residue, or the second lysine residue, or the third lysine residue, or the first and second lysine residues, or the first and third lysine residues, or the second and third lysine residues, or the first, second and third lysine residues of the branched amino acid probe are each Lys q -Lys, (aa3) p -Lys q , Lys q -(aa3) p , [(aa3)-Lys] p and [Lys-(aa3)] p wherein q is a number selected from 0, 1, 2, and 3, p is a number selected from 1, 2, and 3, and (aa3) is an amino acid residue independently selected from Arg, His, Gly, and Ala. In one embodiment, the N-terminal Lys or (aa)3 of the molecule is acetylated at the alpha amino group.

[0164] In certain embodiments of the present disclosure, (aa3) is an amino acid residue independently selected from Gly and Ala. In further embodiments, (aa3) is Gly.

[0165] In one embodiment of the disclosure, the molecule covalently attached to the side chain amino group(s) of the first, second, and / or third alkylamine amino acid residues is acetylated at the alpha amino group of the N-terminal amino acid residue.

[0166] In one embodiment of the present disclosure, the molecules are independently Ac-AAA q -AAA, Ac-(aa3) p -AAA q , Ac-AAA q -(aa3) p , Ac-[(aa3)-AAA] p and Ac-[AAA-(aa3)] p and / or AAA q -AAA, (aa3) p -AAA q, AAA q -(aa3) p , [(aa3)-AAA] p and [AAA-(aa3)] p is selected from the group consisting of:

[0167] In one embodiment of the present disclosure, the molecules are independently Ac-Orn q -Orn, Ac-(aa3) p -Orn q , Ac-Orn q -(aa3) p , Ac-[(aa3)-Orn] p , Ac-[Orn-(aa3)] p , Ac-Orn p -Lys p , Ac-Lys p -Orn p , Ac-[Orn-Lys] p and Ac-[Lys-Orn] p , and / or Orn q -Orn, (aa3) p -Orn q , Orn q -(aa3) p , [(aa3)-Orn] p and [Orn-(aa3)] p , Orn p -Lys p , Lys p -Orn p , [Orn-Lys] p and [Lys-Orn] p is selected from the group consisting of:

[0168] Thus, the molecule, in one embodiment of the present disclosure, is independently Ac-Lys q -Lys, Ac-(aa3) p -Lys q , Ac-Lys q -(aa3) p , Ac-[(aa3)-Lys] p and Ac-[Lys-(aa3)] p , and / or Lys q -Lys, (aa3) p -Lysq , Lys q -(aa3) p , [(aa3)-Lys] p and [Lys-(aa3)] p is selected from the group consisting of:

[0169] In certain embodiments of the present disclosure, the molecule covalently attached to the side chain amino group(s) is Ac-AAA q -AAA or AAA q -AAA, where q is a number selected from 0, 1, 2, and 3.

[0170] Thus, in one embodiment of the present disclosure, the branched amino acid probe consists of 2-9 aminoalkyl amino acid residues, which in one embodiment are individually selected from the group consisting of lysine, D-lysine, ornithine, and D-ornithine.

[0171] In certain embodiments of the present disclosure, the molecule covalently attached to the side chain amino group(s) is Ac-Lys q -Lys or Lys q -Lys, where q is a number selected from 0, 1, 2, and 3.

[0172] Thus, in one embodiment of the present disclosure, the branched amino acid probe consists of 2 to 9 lysine residues.

[0173] In one embodiment of the disclosure, the branched amino acid probe comprises up to 1, 2, 3 or 4 amino acids selected from Arg, His, Gly and Ala(aa3), with the remaining amino acids being aminoalkyl amino acid residues. In another embodiment, the branched amino acid probe comprises up to one Arg residue, and / or up to one His residue, and / or up to one Gly residue, and / or up to one Ala residue.

[0174] In one embodiment of the disclosure, the molecule covalently attached to the side chain amino group(s) of one or more of the first, second and / or third aminoalkyl amino acid residues is AAA, Ac-AAA, AAA-AAA, Ac-AAA-AAA, AAA-AAA-AAA, Ac-AAA-AAA-AAA, AAA-AAA-AAA-AAA, Ac-AAA-AAA-AAA-AAA, AAA-Gly-AAA, Ac-AAA-Gly-AAA, AAA-AAA-Gly, Ac-AAA-AAA-Gly, AAA-Ala-AAA, Ala-A ... and selected from the group consisting of c-AAA-Ala-AAA, AAA-AAA-Ala, Ac-AAA-AAA-Ala, AAA-Ala, Ac-AAA-Ala, AAA-His-AAA, Ac-AAA-His-AAA, AAA-AAA-His, Ac-AAA-AAA-His, AAA-His, Ac-AAA-His, AAA-Arg-AAA, Ac-AAA-Arg-AAA, AAA-AAA-Arg, Ac-AAA-AAA-Arg, AAA-Arg and Ac-AAA-Arg, where AAA is an aminoalkyl amino acid residue as specified herein. Each of the AAA, Gly, Ala, His and Arg amino acid residues above may be in the L or D configuration.

[0175] In one embodiment of the disclosure, the molecule covalently attached to the side chain amino group(s) of one or more of the first, second and / or third aminoalkyl amino acid residues is selected from the group consisting of Lys, Ac-Lys, Lys-Lys, Ac-Lys-Lys, Lys-Lys-Lys, Ac-Lys-Lys-Lys, Lys-Lys-Lys-Lys, Ac-Lys-Lys-Lys, Lys-Gly-Lys, Ac-Lys-Gly-Lys, Lys-Lys-Gly, Ac-Lys-Lys-Gly, Lys-Gly, Ac-Lys-Lys-Gly, Lys-Gly, Ac-Lys-Gly, y, Lys-Ala-Lys, Ac-Lys-Ala-Lys, Lys-Lys-Ala, Ac-Lys-Lys-Ala, Lys-Ala, Ac-Lys-Ala, Lys-His-Lys, Ac-Lys-His-Lys, Lys-Lys-His, Ac- selected from the group consisting of Lys-Lys-His, Lys-His, Ac-Lys-His, Lys-Arg-Lys, Ac-Lys-Arg-Lys, Lys-Lys-Arg, Ac-Lys-Lys-Arg, Lys-Arg and Ac-Lys-Arg.

[0176] In certain embodiments of the present disclosure, the molecule covalently attached to the ε-amino group(s) of one or more of the first, second and / or third lysine residues is selected from the group consisting of Lys, Ac-Lys, Lys-Lys, Ac-Lys-Lys, Lys-Lys-Lys, Ac-Lys-Lys-Lys, Lys-Lys-Lys-Lys, Ac-Lys-Lys-Lys, Lys-Gly-Lys, Ac-Lys-Gly-Lys, Lys-Lys-Gly, Ac-Lys-Lys-Gly, Ly ... ys-Ala-Lys, Ac-Lys-Ala-Lys, Lys-Lys-Ala, Ac-Lys-Lys-Ala, Lys-Ala, Ac-Lys-Ala, Lys-His-Lys, Ac-Lys-His-Lys, Lys-Lys-His, Ac-L selected from the group consisting of ys-Lys-His, Lys-His, Ac-Lys-His, Lys-Arg-Lys, Ac-Lys-Arg-Lys, Lys-Lys-Arg, Ac-Lys-Lys-Arg, Lys-Arg and Ac-Lys-Arg.

[0177] In certain embodiments of the present disclosure, the branched amino acid probe comprises or consists of a first lysine residue selected from Lys and D-Lys, which is optionally N-terminally acetylated or C-terminally amidated, and which has at its ε-amino group: Lys, Ac-Lys, Lys-Lys, Ac-Lys-Lys, Lys-Lys-Lys, Ac-Lys-Lys-Lys, Lys-Lys-Lys-Lys, Ac-Lys-Lys-Lys, Lys-Gly-Lys, Ac-Lys-Gly-Lys, Lys-Lys-Gly, Ac-Lys-Lys, and Ac-Lys-Gly, Lys-Gly, Ac-Lys-Gly, Lys-Ala-Lys, Ac-Lys-Ala-Lys, Lys-Lys-Ala, Ac-Lys-Lys-Ala, Lys-Ala, Ac-Lys-Ala, Lys-His-Lys, Ac-Lys-His-Lys, Lys-Lys-His, Ac-Lys-Lys-His, Lys-His, Ac-Lys-His, Lys-Arg-Lys, Ac-Lys-Arg-Lys, Lys-Lys-Arg, Ac-Lys-Lys-Arg, Lys-Arg and Ac-Lys-Arg.

[0178] In certain embodiments of the present disclosure, the branched amino acid probe comprises or consists of first and second lysine residues selected from Lys and D-Lys, respectively, the second lysine residue being optionally N-terminally acetylated or C-terminally amidated, and i) the first lysine residue, ii) the second lysine residue, or iii) the first and second lysine residues each have one of the following amino acids at the ε-amino group of the lysine residue: Lys, Ac-Lys, Lys-Lys, Ac-Lys-Lys, Lys-Lys-Lys, Ac-Lys-Lys-Lys, Lys-Lys-Lys, Ac-Lys-Lys-Lys, Lys-Gly-Lys, Ac-Lys-G ly-Lys, Lys-Lys-Gly, Ac-Lys-Lys-Gly, Lys-Gly, Ac-Lys-Gly, Lys-Ala-Lys, Ac-Lys-Ala- Lys, Lys-Lys-Ala, Ac-Lys-Lys-Ala, Lys-Ala, Ac-Lys-Ala, Lys-His-Lys, Ac-Lys-His-Lys , Lys-Lys-His, Ac-Lys-Lys-His, Lys-His, Ac-Lys-His, Lys-Arg-Lys, Ac-Lys-Arg-Lys, Lys-Lys-Arg, Ac-Lys-Lys-Arg, Lys-Arg and Ac-Lys-Arg.

[0179] In certain embodiments of the present disclosure, the branched amino acid probe comprises or consists of a first, second and third lysine residue each selected from Lys and D-Lys, the third lysine residue being optionally N-terminally acetylated or C-terminally amidated, and i) the first lysine residue, ii) the second lysine residue, iii) the third lysine residue, iv) the first and second lysine residues, v) the first and third lysine residues, vi) the second and third lysine residues, or vii) the first, second and third lysine residues each having Lys, Ac-Lys, Lys-Lys, Ac-Lys-Lys, Lys-Lys-Lys, Ac-Lys-Lys-Lys, Lys-Lys-Lys-Lys, Ac ... -Lys-Lys-Lys-Lys, Lys-Gly-Lys, Ac-Lys-Gly-Lys, Lys-Lys-Gly, Ac-Lys-Lys-Gly, Lys-Gly, Ac-Lys -Gly, Lys-Ala-Lys, Ac-Lys-Ala-Lys, Lys-Lys-Ala, Ac-Lys-Lys-Ala, Lys-Ala, Ac-Lys-Ala, Lys-His- It is modified by binding a molecule selected from the group consisting of Lys, Ac-Lys-His-Lys, Lys-Lys-His, Ac-Lys-Lys-His, Lys-His, Ac-Lys-His, Lys-Arg-Lys, Ac-Lys-Arg-Lys, Lys-Lys-Arg, Ac-Lys-Lys-Arg, Lys-Arg and Ac-Lys-Arg.

[0180] In one embodiment of the present disclosure, the branched amino acid probe comprises or consists of the formula: Ac-(Ac-Lys-Lys)Lys1- (which is the same as Ac-(Ac-Lys-Lys)Lys-), where Lys1 is a first lysine residue that is acetylated and covalently attached to the N-terminus of a polypeptide as defined herein, and (Ac-Lys-Lys) is a molecule covalently attached to the ε-amino group of the first lysine residue, Lys1.

[0181] In one embodiment of the disclosure, Ac-(Ac-Lys-Lys)Lys- is covalently attached to the N-terminus of a polypeptide defined herein and / or to the side chain amino group of an aminoalkyl amino acid residue within said polypeptide.

[0182] In one embodiment of the disclosure, the branched amino acid probe comprises or consists of the formula: (Ac-Lys-Lys)Lys1-NH2 (same as (Ac-Lys-Lys)Lys1-NH2), where Lys1 is a first lysine residue that is amidated at the C-terminus (-NH2), and (Ac-Lys-Lys) is a molecule attached to the ε-amino group of the first lysine residue, Lys1. In one embodiment of the disclosure, (Ac-Lys-Lys)Lys1-NH2 is attached to the C-terminus of a polypeptide as defined herein.

[0183] In one embodiment of the disclosure, the branched amino acid probe comprises or consists of a formula selected from the group consisting of: (AAA)AAA1-, (AAA-AAA)AAA1-, (AAA-AAA-AAA)AAA1-, (AAA-AAA-AAA-AAA)AAA1-, (AAA-Gly-AAA)AAA1-, (AAA-AAA-Gly)AAA1-, (AAA-Gly)AAA1-, (AAA-Ala-AAA)AAA1-, (AAA-AAA-Ala)AAA1-, (AAA-Ala)AAA1-, (AAA-His-AAA)AAA1-, (AAA-AAA-His)AAA1-, (AAA-His)AAA1-, (AAA-Arg-AAA)AAA1-, (AAA-AAA-Arg)AAA1-, and (AAA-Arg)AAA1-. In one embodiment of the disclosure, the first amino alkyl amino acid residue (AAA1-) is N-terminally acetylated or C-terminally amidated.

[0184] In one embodiment of the present disclosure, the branched amino acid probe is (Lys)Lys1-, (Lys-Lys)Lys1-, (Lys-Lys-Lys)Lys1-, (Lys-Lys-Lys-Lys)Lys1-, (Lys-Gly-Lys)Lys1-, (Lys-Lys-Gly)Lys1-, (Lys-Gly)Lys1-, (Lys-Ala-Lys ... In one embodiment of the present disclosure, the first lysine residue (Lys-Lys-Ala)Lys1-, (Lys-Ala)Lys1-, (Lys-His-Lys)Lys1-, (Lys-Lys-His)Lys1-, (Lys-Arg-Lys)Lys1-, (Lys-Lys-Arg)Lys1-, and (Lys-Arg)Lys1-. In one embodiment of the present disclosure, the first lysine residue (Lys-) is N-terminally acetylated or C-terminally amidated.

[0185] In one embodiment of the present disclosure, the branched amino acid probe is selected from the group consisting of Ac-(Ac-Lys)Lys1-, Ac-(Ac-Lys-Lys)Lys1-, Ac-(Ac-Lys-Lys-Lys)Lys1-, Ac-(Ac-Lys-Lys-Lys-Lys)Lys1-, Ac-(Ac-Lys-Gly-Lys)Lys1-, Ac-(Ac-Lys-Lys-Gly)Lys1-, Ac-(Ac-Lys-Gly)Lys1-, Ac-(Ac-Lys-Ala-Lys)Lys1-, Ac -(Ac-Lys-Lys-Ala)Lys1-, Ac-(Ac-Lys-Ala)Lys1-, Ac-(Ac-Lys-His-Lys)Lys1-, Ac-(Ac-Lys-Lys-His)Lys1-, Ac-(Ac-Lys-His)Lys1-, Ac-(Ac-Lys-Arg-Lys)Lys1-, Ac-(Ac-Lys-Lys-Arg)Lys1-, and Ac-(Ac-Lys-Arg)Lys1-.

[0186] In one embodiment of the present disclosure, the branched amino acid probe is (Ac-Lys)Lysl-NH2, (Ac-Lys-Lys)Lysl-NH2, (Ac-Lys-Lys-Lys)Lysl-NH2, (Ac-Lys-Lys-Lys-Lys)Lysl-NH2, (Ac-Lys-Gly-Lys)Lysl-NH2, (Ac-Lys-Lys-Gly)Lysl-NH2, (Ac-Lys-Gly)Lysl-NH2, (Ac-Lys-Ala-Lys)Lysl-NH2, (Ac-Lys-Lys-Ala)Lysl-NH 2, comprising or consisting of a formula selected from the group consisting of (Ac-Lys-Ala)Lysl-NH2, (Ac-Lys-His-Lys)Lysl-NH2, (Ac-Lys-Lys-His)Lysl-NH2, (Ac-Lys-Arg-Lys)Lysl-NH2, (Ac-Lys-Lys-Arg)Lysl-NH2, and (Ac-Lys-Arg)Lysl-NH2.

[0187] More specifically, in one embodiment of the present disclosure, the branched amino acid probe comprises or consists of a formula selected from the group consisting of Ac-(Ac-Lys)Lys1-, Ac-(Ac-Lys-Lys)Lys1-, Ac-(Ac-Lys-Lys-Lys)Lys1-, Ac-(Ac-Lys-Lys-Lys-Lys)Lys1-, Ac-(Ac-Lys-Gly-Lys)Lys1-, Ac-(Ac-Lys-Lys-Gly)Lys1- and Ac-(Ac-Lys-Gly)Lys1-.

[0188] In one embodiment of the present disclosure, the branched amino acid probe comprises or consists of the formula: Ac-(Ac-Lys)Lys2-Lys1-, where Lys1 is a first lysine residue, Lys2 is a second lysine residue that is acetylated and covalently attached to Lys1 via a peptide bond, and (Ac-Lys) is a molecule covalently attached to the ε-amino group of the second lysine residue, Lys2.

[0189] In one embodiment of the present disclosure, the branched amino acid probe comprises or consists of the formula: Ac-Lys2-(Ac-Lys)Lys1-, where the molecule (Ac-Lys) is covalently attached to the ε-amino group of the first lysine residue, Lys1.

[0190] In one embodiment of the present disclosure, the branched amino acid probe(s) include Ac-(Ac-Lys)Lys-Lys-, (Ac-Lys)Lys-Lys-, Ac-(Lys)Lys-Lys-, (Lys)Lys-Lys-, (Ac-Lys)Lys-Lys-NH2, (Lys)Lys-Lys-NH2, Ac-Lys-(Ac-Lys)Lys-, Lys-(Ac-Lys)Lys-, Ac-Lys-(Lys)Lys-, Lys-(Lys)Lys- Lys-(Ac-Lys)Lys-NH2, Lys-(Lys)Lys-NH2, Ac-(Ac-Lys-Lys)-Lys-, (Ac-Lys-Lys)-Lys-, Ac-(Lys-Lys)-Lys- and (Lys-Lys)-Lys- (Ac-Lys-Lys)-Lys-NH2, and (Lys-Lys)-Lys-NH2.

[0191] In one embodiment of the present disclosure, the branched amino acid probe(s) is selected from the group consisting of Ac-(Ac-Lys)Lys-, Ac-(Lys)Lys-, (Ac-Lys)Lys-NH2, (Lys)Lys-NH2 and (Lys)Lys-.

[0192] In one embodiment of the present disclosure, the branched amino acid probes are Ac-(Ac-Lys)Lys2-Lys1-, Ac-(Ac-Lys-Lys)Lys2-Lys1-, Ac-(Ac-Lys-Gly)Lys2-Lys1-, Ac-(Ac-Lys-Lys-Lys)Lys2-Lys1-, Ac-(Ac-Lys-Lys-Lys-Lys)Lys2-Lys1-, Ac-Lys2-(Ac-Lys)-Lys1-, Ac-Lys2-(Ac-Lys)-Lys1-, Ac-Lys2-(Ac-Lys)-Lys1-, A c-Lys2-(Ac-Lys-Gly)-Lys1-, Ac-Lys2-(Ac-Lys-Lys-Lys)-Lys1-, Ac-Lys2-(Ac-Lys-Lys-Lys-Lys)-Lys1-, Ac-(Ac-Lys)Lys2 -(Ac-Lys-)-Lys1-, Ac-(Ac-Lys)Lys2-(Ac-Lys-Lys-)-Lys1-, and Ac-(Ac-Lys-Lys)Lys2-(Ac-Lys-Lys-)-Lys1-.

[0193] More specifically, in one embodiment of the present disclosure, the branched amino acid probe is Ac-(Ac-Lys)Lys2-Lys1-, Ac-(Ac-Lys-Lys)Lys2-Lys1-, Ac-(Ac-Lys-Gly)Lys2-Lys1-, Ac-Lys2-(Ac-Lys)-Lys1-, Ac-Lys2-(Ac-Lys-Lys)-Lys2-Lys s1-, Ac-Lys2-(Ac-Lys-Gly)-Lys1-, Ac-(Ac-Lys)Lys2-(Ac-Lys-)-Lys1-, Ac-(Ac-Lys)Lys2-(Ac-Lys-Lys-)-Lys1-, and Ac-(Ac-Lys-Lys)Lys2-(Ac-Lys-Lys-)-Lys1-.

[0194] In one embodiment of the present disclosure, the branched amino acid probe is Ac-Lys3-Lys 2-(Ac-Lys)Lys1-, Ac-Lys3-(Ac-Lys)Lys2-Lys1-, Ac-(Ac-Lys)Lys3-Lys2-Lys1-, Ac-Lys3-(Ac-Lys)Lys2-(Ac- Lys)Lys1-, Ac-(Ac-Lys)Lys3-(Ac-Lys)Lys2-Lys1-, and Ac-(Ac-Lys)Lys3-Lys2-(Ac-Lys)Lys1-.

[0195] In certain embodiments of the present disclosure, the branched amino acid probes are Ac-(Ac-Lys)Lys1-, Ac-(Ac-Lys-Lys)Lys1-, Ac-(Ac-Lys-Lys-Lys)Lys1-, Ac-(Ac-Lys-Lys-Lys-Lys)Lys1-, Ac-(Ac-Lys-Gly-Lys)Lys1-, Ac-(Ac-Lys-Lys-Gly)Lys1-, Ac-(Ac-Lys-Gly)Lys1-, Ac-(Ac-Lys-Lys2-Lys1-, Ac-(Ac-Lys-Lys) ...2-Lys2-Lys2-Lys2-Lys2-Lys2-Lys2-Lys2-Lys2-Lys2-Lys2-Lys2-Lys2-Lys2-L ys2-Lys1-, Ac-(Ac-Lys-Gly)Lys2-Lys1-, Ac-Lys2-(Ac-Lys)-Lys1-, Ac-Lys2-(Ac-Lys-Lys)-Lys1-, Ac-Lys2-(Ac-Lys-Gly)-Lys1-, A c-(Ac-Lys)Lys2-(Ac-Lys-)-Lys1-, Ac-(Ac-Lys)Lys2-(Ac-Lys-Lys-)-Lys1-, Ac-(Ac-Lys-Lys)Lys2-(Ac-Lys-Lys-)-Lys1-, Ac-Lys3- Lys 2- The branched amino acid probe is selected from the group consisting of (Ac-Lys)Lys1-, Ac-Lys3-(Ac-Lys)Lys2-Lys1-, Ac-(Ac-Lys)Lys3-Lys2-Lys1-, Ac-Lys3-(Ac-Lys)Lys2-(Ac-Lys)Lys1-, Ac-(Ac-Lys)Lys3-(Ac-Lys)Lys2-Lys1-, and Ac-(Ac-Lys)Lys3-Lys2-(Ac-Lys)Lys 1. In one embodiment, the branched amino acid probe is covalently attached to the N-terminus of a polypeptide as defined herein and / or to the side chain amino group of an aminoalkyl amino acid residue within the polypeptide.

[0196] In certain embodiments of the present disclosure, the branched amino acid probe consists of two or three lysine residues (selected from Lys and D-Lys).

[0197] In certain embodiments of the present disclosure, the branched amino acid probe consists of three lysine residues. In another embodiment, the branched amino acid probe consists of two lysine residues.

[0198] In certain embodiments of the present disclosure, the branched amino acid probe consists of first and second lysine residues selected from Lys and D-Lys, one or both of which are modified by attachment to the epsilon amino group of the first and / or second lysine residues a lysine residue selected from Lys and D-Lys, each of which is optionally acetylated at the alpha amino group.

[0199] In certain embodiments of the present disclosure, the branched amino acid probe consists of a first lysine residue selected from Lys and D-Lys, which is modified by attaching to the epsilon-amino group of the first lysine residue two lysine residues selected from Lys and D-Lys, each of which is optionally acetylated at the alpha amino group.

[0200] Linkage of BAP to polypeptides According to the present disclosure, the first aminoalkyl amino acid residue of each of the one or more branched amino acid probes is covalently attached to the N-terminus of a polypeptide or polypeptide analogue as defined herein, covalently attached to the C-terminus of the polypeptide or polypeptide analogue, and / or covalently attached to the side chain amino group of an aminoalkyl amino acid residue within the polypeptide or polypeptide analogue.

[0201] Attachment of one or more branched amino acid probes to a polypeptide or polypeptide analogue as defined herein results in a polypeptide / BAP conjugate, such as a polypeptide analogue / BAP conjugate.

[0202] The term covalently linked to the N-terminus of the polypeptide or polypeptide analog means that the first aminoalkyl amino acid residue of the branched amino acid probe is covalently linked to the alpha amino group of the most N-terminal amino acid residue of the polypeptide or polypeptide analog.

[0203] The term covalently linked to the C-terminus of the polypeptide or polypeptide analog means that the alpha amino group of the first aminoalkyl amino acid residue of the branched amino acid probe is covalently linked to the most C-terminal amino acid residue of the polypeptide or polypeptide analog.

[0204] It is further understood that the branched amino acid probe in one embodiment of the present disclosure is covalently attached to the side chain amino group of an aminoalkyl amino acid residue within the polypeptide or polypeptide analog of interest.

[0205] In one particular embodiment of the disclosure, the aminoalkyl amino acid residue in the polypeptide or polypeptide analog sequence is selected from the group consisting of ornithine and lysine residues, hi one particular embodiment of the disclosure, the aminoalkyl amino acid residue in the peptide sequence is a lysine residue.

[0206] In one embodiment of the disclosure, the first aminoalkyl amino acid residue of the branched amino acid probe is covalently linked to the δ-amino group of an ornithine residue in the polypeptide or polypeptide analog or to the ε-amino group of a lysine residue in the polypeptide or polypeptide analog.

[0207] In one embodiment of the present disclosure, the first aminoalkyl amino acid residue of the branched amino acid probe is covalently attached to the epsilon-amino group of a lysine residue in the polypeptide or polypeptide analog of interest.

[0208] An aminoalkyl amino acid residue within the peptide sequence is understood to mean that the aminoalkyl amino acid residue does not form part of the branched amino acid probe itself, but is a residue that is present within the existing amino acid sequence of the polypeptide or polypeptide analogue. The aminoalkyl amino acid residue may be located at any position in the polypeptide or polypeptide analogue.

[0209] According to the present disclosure, a polypeptide analog comprising one or more branched amino acid probes means that in one embodiment the polypeptide analog comprises one branched amino acid probe, such as two branched amino acid probes, three branched amino acid probes, e.g., four branched amino acid probes, five branched amino acid probes, e.g., six branched amino acid probes.

[0210] In principle, the polypeptide analogs of the present disclosure can include any number of branched amino acid probes, provided that the polypeptide analog can be covalently linked to a peptide (at the N-terminus, C-terminus, and / or to one or more aminoalkyl amino acid residues within the polypeptide).

[0211] In one embodiment of the present disclosure, the polypeptide analog comprises one branched amino acid probe.

[0212] In one embodiment of the present disclosure, the polypeptide analog comprises one branched amino acid probe, which is covalently attached to the N-terminus of the polypeptide.

[0213] In one embodiment of the disclosure, the polypeptide analog comprises one branched amino acid probe, which is covalently attached to the C-terminus of the polypeptide.

[0214] In one embodiment of the disclosure, a polypeptide analog comprises one branched amino acid probe that is covalently attached to the side chain amino group of an aminoalkyl amino acid residue in the polypeptide.

[0215] In one embodiment of the present disclosure, a polypeptide analog comprises more than one (two or more) branched amino acid probe(s). In embodiments in which a polypeptide analog comprises more than one branched amino acid probe, it is understood that the more than one branched amino acid probes may be individually the same (identical) or different (non-identical).

[0216] In one embodiment of the present disclosure, the polypeptide analog comprises two branched amino acid probes.

[0217] In one embodiment of the present disclosure, a polypeptide analog comprises two branched amino acid probes, one branched amino acid probe covalently attached to the N-terminus of the polypeptide and another branched amino acid probe covalently attached to the C-terminus of the polypeptide.

[0218] In one embodiment of the present disclosure, a polypeptide analog comprises two branched amino acid probes, one covalently attached to the N-terminus of a polypeptide and another covalently attached to the side chain amino group of an aminoalkyl amino acid residue within the polypeptide.

[0219] In one embodiment of the present disclosure, a polypeptide analog comprises two branched amino acid probes, one covalently attached to the C-terminus of a polypeptide and another covalently attached to the side chain amino group of an aminoalkyl amino acid residue within the polypeptide.

[0220] In one embodiment of the disclosure, a polypeptide analog comprises two branched amino acid probes, each of which is covalently attached to the side chain amino group of a different (or distinct) aminoalkyl amino acid residue within the polypeptide.

[0221] In one embodiment of the present disclosure, the polypeptide analog comprises three branched amino acid probes.

[0222] In one embodiment of the disclosure, a polypeptide analog comprises three branched amino acid probes, a first branched amino acid probe covalently attached to the N-terminus of the polypeptide, a second branched amino acid probe covalently attached to the C-terminus of the polypeptide, and a third branched amino acid probe covalently attached to the side chain amino group of an aminoalkyl amino acid residue in the polypeptide analog.

[0223] In one embodiment of the present disclosure, a polypeptide analog comprises three branched amino acid probes, a first branched amino acid probe covalently attached to the N-terminus of a polypeptide, and a second and third branched amino acid probes each covalently attached to the side chain amino group of a different aminoalkyl amino acid residue within the polypeptide.

[0224] In one embodiment of the present disclosure, a polypeptide analog comprises three branched amino acid probes, a first branched amino acid probe covalently attached to the C-terminus of a polypeptide, and a second and third branched amino acid probes each covalently attached to the side chain amino group of a different aminoalkyl amino acid residue within the polypeptide.

[0225] Treatment method In one embodiment, the present disclosure relates to polypeptide analogs of annexin A1, conjugated forms of the polypeptide analogs, and compositions comprising the polypeptide analogs of annexin A1 or their conjugated forms for use as medicaments and for treating ischemic and / or inflammatory conditions.

[0226] It is one aspect of the present disclosure to provide a polypeptide or polypeptide analog selected from the group consisting of SEQ ID NO:2 to SEQ ID NO:24, SEQ ID NO:25 to SEQ ID NO:47, and SEQ ID NO:48 to SEQ ID NO:70, or a functional variant thereof, for use as a pharmaceutical.

[0227] Furthermore, one aspect of the present disclosure relates to a polypeptide or polypeptide analog selected from the group consisting of SEQ ID NO:2 to SEQ ID NO:24, SEQ ID NO:25 to SEQ ID NO:47, and SEQ ID NO:48 to SEQ ID NO:70, or a functional variant thereof, for use in a method for treating an ischemic and / or inflammatory condition.

[0228] In certain embodiments, a polypeptide or polypeptide analog selected from the group consisting of: AMVSEFLKQAWFIENEEQEYVQTLKSSK (SEQ ID NO:50), AMVSEFLKQAWFIENEEQEYVQTLKSSKG (SEQ ID NO:51), AMVSEFLKQAWFIENEEQEYVQTLKSSKGG (SEQ ID NO:52), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGP (SEQ ID NO:53), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPG (SEQ ID NO:54), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGS (SEQ ID NO:55), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSA (SEQ ID NO:56), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAV (SEQ ID NO:57), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVS (SEQ ID NO:58), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSP (SEQ ID NO:59), and AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPY (SEQ ID NO: 60), or a functional variant thereof is provided.

[0229] In a preferred embodiment, the polypeptide or polypeptide analogue for use in the treatment of an ischemic and / or inflammatory condition is selected from the group consisting of: AMVSEFLKQAWFIENEEQEYVQTLKSSK (SEQ ID NO:50), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGS (SEQ ID NO:55), and AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPY (SEQ ID NO: 60), or a functional variant thereof.

[0230] Functional variants are disclosed elsewhere herein.

[0231] The present disclosure further relates to a polypeptide or polypeptide analog selected from the group consisting of SEQ ID NO:2 to SEQ ID NO:24, SEQ ID NO:25 to SEQ ID NO:47, and SEQ ID NO:48 to SEQ ID NO:70, or a functional variant thereof, for use in the manufacture of a medicament for treating an ischemic and / or inflammatory condition.

[0232] In another aspect, the disclosure provides a method for the treatment, prevention or alleviation of an ischemic and / or inflammatory condition, such a method according to the disclosure, in one embodiment, comprises one or more steps of administering or delivering to an individual in need thereof an effective amount of a polypeptide or polypeptide analog selected from the group consisting of SEQ ID NO:2-SEQ ID NO:24, SEQ ID NO:25-SEQ ID NO:47, and SEQ ID NO:48-SEQ ID NO:70, or a functional variant thereof, or a pharmaceutical composition comprising one or more such polypeptides or polypeptide analogs, as disclosed herein.

[0233] An individual in need as referred to herein is, in one embodiment, an individual who will benefit from administration of a polypeptide or polypeptide analog disclosed herein, or a pharmaceutical composition comprising said polypeptide or polypeptide analog according to the present disclosure. Such an individual, in one embodiment, suffers from or is at risk of suffering from a disease or condition. An individual, in one embodiment, is any human being, male or female, infant, middle-aged or elderly. The disorder to be treated or prevented in an individual, in one embodiment, is related to the age of the individual, the general health of the individual, the drugs used to treat the individual, and whether the individual has a history of suffering from a disease or disorder that may have induced or induced the condition in the individual.

[0234] The terms "treatment" and "treating" as used herein refer to the management and care of a patient for the purpose of combating a condition, disease or disorder. The terms are intended to include the full range of treatments, such as administration of a polypeptide analog of the present disclosure for a given condition from which the patient is suffering, with the aim of alleviating or relieving symptoms or complications, delaying the progression of a condition, partially arresting clinical symptoms, disease or disorder, curing or eliminating a condition, disease or disorder, and / or preventing or reducing the risk of contracting a condition, disease or disorder, and "preventing" or "prevention" is understood to refer to the management and care of a patient for the purpose of preventing the onset of a condition, disease or disorder, including administration of an active compound to prevent or reduce the risk of developing a condition or complication. The patient to be treated is preferably a mammal, particularly a human. However, treatment of animals such as mice, rats, dogs, cats, cows, horses, sheep, pigs, etc. is also within the scope of the present disclosure. Patients treated by the present disclosure can be of various ages, for example, adults, children, children under 16 years old, children between 6 and 16 years old, children between 2 and 16 years old, children between 2 months and 6 years old, or children between 2 months and 5 years old.

[0235] As used herein, references to "inflammation" or "inflammatory response or disease or condition" refer to any inflammatory response or disease, including, but not limited to, inflammation of the eye, gout, gouty arthritis, rheumatoid arthritis, asthma, reperfusion injury or damage, stroke, myocardial infarction, septic shock, or inflammatory skin disorders, such as psoriasis or eczema.

[0236] Medical Indication In one aspect, the disclosure provides a polypeptide or polypeptide analogue disclosed herein for use in treating an ischemic and / or inflammatory condition.

[0237] In some embodiments of the present disclosure, the treatment is prophylactic, ameliorative, and / or curative.

[0238] In some embodiments of the present disclosure, the ischemic and / or inflammatory condition is acute, subacute, or chronic.

[0239] In one embodiment of the disclosure, there is provided a polypeptide or polypeptide analogue as disclosed herein for use in treating an ischemic and / or inflammatory condition in tissue of one or more organs of a mammal.

[0240] In some embodiments of the disclosure, the mammal is a homo sapiens.

[0241] In some embodiments of the present disclosure, the organ is selected from the group consisting of kidney, liver, brain, heart, muscle, bone marrow, skin, skeleton, lung, airway, spleen, exocrine glands, bladder, endocrine glands, reproductive organs including the vaginal tract, eye, ear, vascular system, gastrointestinal tract including the small intestine, colon, rectum, anal canal, and prostate.

[0242] In some embodiments of the present disclosure, the ischemic condition is secondary ischemia.

[0243] In some embodiments of the present disclosure, the ischemia is secondary ischemia, which results from stroke, injury, septic shock, systemic hypotension, cardiac arrest due to heart attack, cardiac arrhythmia, atherosclerotic disease with thrombosis, embolism of the blood vessels of the heart or any organ, vasospasm, aortic aneurysm or aneurysm of other organs, coronary artery stenosis, myocardial infarction, angina pectoris, pericarditis, myocarditis, myxedema, or endocarditis.

[0244] The aortic aneurysm, in one embodiment, is a thoracic or abdominal or dissecting aortic aneurysm. Systemic hypotension, in one embodiment, is hypotension due to cardiac disease, hypotension due to systemic disease including infection or an allergic reaction, or hypotension due to one or more toxic compounds or toxins(es) or drug(s).

[0245] In some embodiments of the present disclosure, the ischemic condition is myocardial ischemia.

[0246] In some embodiments of the present disclosure, the ischemic and / or inflammatory condition is associated with surgery, such as major surgery, or with organ transplantation, such as solid organ transplantation.

[0247] In some embodiments of the present disclosure, the ischemic and / or inflammatory condition is selected from the group consisting of postoperative systemic inflammatory response syndrome (SIRS) and postoperative organ dysfunction, such as postoperative renal failure including, for example, acute kidney injury (AKI), nephrotoxicity and / or chronic renal failure (CRF).

[0248] In some embodiments of the present disclosure, the ischemic and / or inflammatory condition is reperfusion injury.

[0249] In some embodiments of the present disclosure, the inflammatory disease is selected from the group consisting of arthropathy (joint disease), rheumatoid arthritis (RA), gout, inflammatory diseases of the gastrointestinal system, and multiple sclerosis.

[0250] In some embodiments of the present disclosure, the ischemic and / or inflammatory condition is liver inflammation. In one embodiment of the present disclosure, the inflammatory disease is non-alcoholic steatohepatitis (NASH).

[0251] In some embodiments of the present disclosure, the ischemic and / or inflammatory condition in the tissue of one or more organs is caused by cardiac arrhythmia. In one embodiment of the present disclosure, the cardiac arrhythmia is a primary disease or is secondary to another condition in the individual, including acute infections, particularly those affecting the lungs, pulmonary embolism, hypotension, shock, anoxemia, and anemia.

[0252] Cardiac arrhythmias include ventricular or supraventricular tachyarrhythmias, atrioventricular block, insight point disease, Borff-Parkinson-White syndrome, Leneglés disease, Lew disease, and any syndrome involving abnormal myocardial connections between the atria and ventricles.

[0253] In some embodiments of the present disclosure, secondary ischemia may also be observed in association with a variety of other diseases and conditions, including, but not limited to, diabetes mellitus, hyperlipidemia, thromboangiitis obliterans, Takayasu's syndrome, temporal arteritis, mucocutaneous lymph node syndrome (Kawasaki's disease), cardiovascular coleoptosis, connective tissue disorders such as Raynaud's disease, vascular trauma including iatrogenic trauma such as acne imprinting, cannulation, conditions with elevated fasting levels of LDL cholesterol, triglycerides and / or HDL cholesterol, retroperitoneal fibrosis, rheumatic diseases, systemic lupus erythematosus, lymphocytic polyarteritis, scleroderma, polymyositis, dermatomyositis, rheumatoid arthritis, neuromuscular disorders such as Duchenne's progressive muscular dystrophy, Friedrich's ataxia, and myotonic dystrophy, anaphylaxis, serum sickness, hemolytic anemia, allergy, and allergic agranulocytosis. In one embodiment, the polypeptides and polypeptide analogs of the present disclosure are also useful in the treatment or prevention of the above conditions.

[0254] In some embodiments of the present disclosure, the condition to be treated is due to cancer, including acute leukemia of any origin, chronic myeloid leukemia, chronic lymphocytic leukemia, Hodgkin's disease, lymphosarcoma, myeloma, metastatic cancer, or a pre-malignant disorder affecting an organ, e.g., the respiratory system, including the lungs, bronchioles, upper respiratory tract, and / or the heart and / or the kidneys and / or the gastrointestinal system. In some embodiments of the present disclosure, the polypeptides or polypeptide analogs disclosed herein are used in the treatment or prevention of the above conditions.

[0255] In some embodiments of the present disclosure, the ischemic and / or inflammatory condition in tissue of one or more organs is caused by physical trauma, including electromagnetic radiation.

[0256] One aspect of the present disclosure provides a method for the treatment of an ischemic and / or inflammatory condition comprising administering to an individual in need thereof an effective amount of a polypeptide or polypeptide analog disclosed herein.

[0257] Surgery and transplantation Major surgical interventions, including cardiac surgery, abdominal surgery, aortic and other major vascular surgery, and organ transplants such as lung or heart or combined lung and heart transplants, liver transplants or kidney transplants, induce systemic inflammatory responses (SIR; or systemic inflammatory response syndrome SIRS) and are associated with postoperative organ dysfunction, including the development of renal failure.

[0258] Renal failure is the result of the reduced blood flow that occurs during SIR and surgical intervention. The result is postoperative acute kidney injury (KI), which in most cases worsens to chronic renal failure. Currently, there is no efficient treatment to prevent the development of renal failure. Postoperative renal failure can be defined as a greater than 25% decrease in glomerular filtration rate (GFR) present 3 months after surgical intervention.

[0259] Major cardiac surgery such as repair of one or more heart valves, cardiac artery bypass graft (CABG), aortic root surgery, or surgery on the aortic bifurcation involving the common carotid artery, or concomitant cardiac surgery such as valve(s) replacement and CABG and / or aortic root surgery, are associated with the development of renal impairment and, when present, increased morbidity and mortality.

[0260] In some embodiments of the disclosure, treatment with a polypeptide or polypeptide analog disclosed herein reduces the extent of renal damage, for example, by reducing the decline in postoperative GFR, reducing the extent of the postoperative increase in serum creatinine or cystatin C, or the more immediate increase in urinary excretion of AKI markers NGAL, IL18, or KIM-1, and / or reducing the extent of postoperative SIR (e.g., by reducing circulating levels of IL-6 and other pro-inflammatory markers).

[0261] Lung transplantation (LTX) is the definitive treatment for end-stage lung disease. The major challenges associated with LTX are the shortage of donors, acute and chronic rejection of the transplanted lung, and side effects of immunosuppressive treatment, including the development of chronic renal failure (CRF).

[0262] Although there have been good advances in the treatment of acute rejection with newer immunosuppressants, resulting in fewer episodes of acute rejection within the first year, less organ loss, fewer side effects, fewer infections, and less invasive monitoring methods, the management of chronic organ rejection has not improved significantly and the half-life for the time that 50% of patients survive has improved only slightly over the past 20 years, approximately 70%.

[0263] The side effects of immunosuppressive treatment are dominated by two main challenges: nephrotoxicity and post-transplant lymphoproliferative disease (PTLD), the latter of which can be considered as a consequence of the degree of immunosuppression required to avoid chronic organ rejection - "too much" side effects will keep rejection at a distance but cause infections and PTLD, while "too little" side effects will increase the risk that the patient will reject the graft. Nephrotoxicity and the development of CRF remain significant problems despite extensive research in the past 30 years. Five years after LTX, no patient maintains normal renal function, and even 20% of long-term survivors will end up with a kidney transplant.

[0264] Calcineurin inhibitor therapy (tacrolimus, cyclosporine A) is the cornerstone of the immunosuppressive treatment strategy for successful solid organ transplantation. The limiting factor in using calcineurin inhibitors is acute and chronic irreversible nephrotoxicity. Recent data show that renal function (measured as a reduction in GFR) is reduced by 40% within the first 14 days after LTX and that this reduction is irreversible.

[0265] Heart transplantation (HTX) is the definitive treatment for end-stage heart failure. As for LTX, the main challenges associated with HTX are the shortage of donors, acute and chronic rejection of the transplanted heart, and side effects of immunosuppressive treatment, including the development of CRF. As with LTX, the number of patients who retain renal function over time is limited or absent, and as with LTX, the major reduction in renal function is already present 2-4 weeks after transplantation.

[0266] This dramatic effect on renal function seen after LTX and HTX is likely not caused by calcineurin inhibitor treatment alone, but is the end result of surgical and anesthetic trauma combined with organ ischemia and the side effects of antibiotics, antivirals, antifungals and immunosuppressants. Thus, in one embodiment, pharmacological intervention using the disclosed polypeptides or polypeptide analogs will reduce the extent of renal damage associated with organ transplants such as LTX and HTX.

[0267] Thus, surgery, as outlined in detail herein above, including organ transplantation, may be a cause of secondary ischemia.

[0268] Thus, the present disclosure in one embodiment relates to a polypeptide or polypeptide analog as disclosed herein for use in treating an ischemic and / or inflammatory condition in a tissue of one or more organs of a mammal, said ischemic and / or inflammatory condition being associated with surgery, in one embodiment of the disclosure said surgery being a major surgery or major surgical intervention.

[0269] In one embodiment of the present disclosure, the surgery is selected from the group consisting of cardiac surgery, abdominal surgery, surgery on the aorta and / or other major blood vessels, repair of one or more heart valves, cardiac artery bypass graft (CABG), surgery on the aortic root or aortic branches including the common carotid artery, and combined cardiac surgery such as valve(s) replacement and CABG and / or aortic root surgery.

[0270] In one embodiment of the present disclosure, the procedure involves the surgical insertion of an implant, device, graft, prosthesis, or other biological compound or device that is inserted by surgery.

[0271] In one embodiment of the disclosure, the major surgery comprises an organ transplant. Thus, in one embodiment, the disclosure relates to a polypeptide or polypeptide analog as disclosed herein for use in treating an ischemic and / or inflammatory condition in tissue of one or more organs of a mammal, the ischemic and / or inflammatory condition being associated with an organ transplant. In one embodiment of the disclosure, the organ transplant is a solid organ transplant.

[0272] In one embodiment of the present disclosure, the solid organ transplant is a heart transplant, a lung transplant, a combined heart and lung transplant, a liver transplant, or a renal (kidney) transplant.

[0273] In another embodiment, the present disclosure relates to a polypeptide or polypeptide analog as disclosed herein for use in the treatment of post-operative renal failure, such as post-operative systemic inflammatory response syndrome (SIRS), post-operative organ dysfunction and / or acute kidney injury (AKI), nephrotoxicity and / or chronic renal failure (CRF).

[0274] The present disclosure relates, in one embodiment, to a polypeptide or polypeptide analog as disclosed herein for reducing the extent of renal damage associated with major surgery, in one embodiment, organ transplantation.

[0275] Reperfusion injury is tissue damage that occurs when blood supply is returned to tissue after a period of ischemia or oxygen deprivation. The absence of oxygen and nutrients from the blood during the ischemic period creates a condition in which restoration of circulation leads to inflammation and oxidative damage through the induction of oxidative stress rather than restoration of normal function.

[0276] Reperfusion injury can occur in association with surgery, such as major surgical interventions including organ transplantation. It is a major concern when performing liver transplants and during cardiac surgery.

[0277] In certain embodiments of the present disclosure, the ischemic and / or inflammatory condition in the tissue of one or more organs is associated with reperfusion injury. Thus, in one embodiment, the present disclosure relates to a polypeptide or polypeptide analog as disclosed herein for use in treating an ischemic and / or inflammatory condition in the tissue of one or more organs of a mammal, wherein the ischemic and / or inflammatory condition is associated with reperfusion injury.

[0278] In some embodiments of the disclosure, a polypeptide, polypeptide analog, or composition of the disclosure is to be administered before and / or during surgery and / or organ transplantation.

[0279] In some embodiments of the present disclosure, the ischemic and / or inflammatory conditions in tissues of one or more organs described herein are caused by toxin- or drug-induced cell, tissue or organ failure.

[0280] Toxins and Drugs The present disclosure relates, in one embodiment, to a polypeptide or polypeptide analog as disclosed herein for use in treating an ischemic and / or inflammatory condition in tissue of one or more organs of a mammal, said ischemic and / or inflammatory condition caused (or induced) by toxin or drug-induced cell, tissue or organ failure.

[0281] The above drugs include cisplatin, carboplatin, dacarbazine, procarbazine, altretamine, semustine, lomustine, carmustine, busulfan, thiotepa, melphalan, cyclophosphamide, chlorambucil, mechlorethamine, azacitidine, cladribine, cytarabine, fludarabine, fluorouracil, mercaptopurine, methotrexate, thioguanine, allopurinol, bleomycin, dactinomycin, daunorubicin, docetaxel, doxorubicin (adriamycin), etoposide, idarubicin, irinotecan, Antibiotics as aminoglycosides, including mitomycin, paclitaxel, primycin, topotecan, vinblastine, vincristine, vinorelbine, amsacrine, asparaginase, hydroxyurea, mititane, mitoxantrone, streptomycin, neomycin, kanamycin, amikacin, gentamicin, tobramycin, sisomicin, and nitylmcin; immunosuppressant compounds such as cyclosporine; cancer chemotherapeutic agents including tricyclic antidepressants, lithium salts, prenylamines, and phenothiidine derivatives.

[0282] Inflammatory conditions Inflammation is the body's local defense response against pathogens and injury. Immune cells and soluble factors participate in this process to neutralize damaging agents and initiate tissue repair to restore homeostasis. Loss of regulation of these mechanisms impedes the final resolution of the inflammatory process, resulting in chronic inflammation. Chronic inflammation is of great importance in modern medicine today, as it contributes to the pathogenesis of the most important diseases of industrialized society, including atherosclerosis, acute and chronic heart failure, cancer, diabetes, and obesity-related diseases. Recent insights into endogenous anti-inflammatory pathways have identified a large number of natural anti-inflammatory and pro-resolving molecules and pathways suitable for pharmacological intervention, which makes it possible to develop drugs that mimic the natural process of resolving inflammation. Among these natural anti-inflammatory and pro-resolving pathways is Annexin A1, which acts via FPR2 stimulation.

[0283] The immunomodulatory effects of FPR-2 agonists are exerted through the inhibition of inflammatory mediators and by inhibition of inflammatory cell migration. FPR-2 agonists exert these effects on a variety of cells, including monocytes, macrophages, T cell subtypes, endothelial cells, and epithelial cells.

[0284] Joint diseases such as rheumatoid arthritis (RA) and gout are characterized by episodes with acute exacerbations; in RA, exacerbations (often described as flares) typically occur in addition to chronic symptoms and despite intensive pharmacological treatment. A similar pattern is seen in gout, with the main difference being that most gout patients are asymptomatic between exacerbations. In both conditions, prominent neutrophil infiltration into the synovium and joint fluid is the main pathological feature of exacerbations. The most important proinflammatory effectors involved include IL-1β, TNF-α, IL-6, IL-8, and COX-2. Resolution of acute exacerbations to avoid the development or worsening of chronic inflammation involves activation of macrophages to phagocytose apoptotic neutrophils.

[0285] Therefore, in one embodiment of the present disclosure, it would be attractive to apply treatment with the polypeptide or polypeptide analog disclosed herein to joint disease, not to reduce the severity of exacerbations in existing diseases, at least as rheumatoid arthritis flare has a major clinical impact.However, it is not only joint disease that is associated with exacerbations of symptoms.Neurodegenerative diseases such as multiple sclerosis have relapsing exacerbations, and in one embodiment of the present disclosure, treatment with the polypeptide or polypeptide analog disclosed herein can reduce symptoms and ultimately reduce the overall decline in the functional level of patients with respect to joint disease.

[0286] The present disclosure relates in one embodiment to a polypeptide or polypeptide analog as disclosed herein for use in treating an inflammatory condition in tissue of one or more organs of a mammal, wherein said ischemic and / or inflammatory condition is an inflammatory disease.

[0287] In one embodiment of the disclosure, the inflammatory disease is arthritis. In one embodiment of the disclosure, the inflammatory disease is arthropathy (diseases of the joints), arthritis (including diseases related to arthritis), osteoarthritis, rheumatoid arthritis; spondyloarthropathy (e.g., ankylosing spondylitis), reactive arthritis (including post-rheumatic fever arthritis), juvenile chronic arthritis including Henoch-Schanlein purpura, Reiner's disease, Still's disease, juvenile rheumatoid arthritis, juvenile ankylosing spondylitis, psoriasis, osteoarthritis, hypermobile arthropathy, congenital dysplasia, femoral metaphysis forming runway, osteoarthritis secondary to Perthes' disease, intra-articular fractures, meniscal dissections. The conditions for which the present invention is concerned are selected from the group consisting of osteotomy, obesity, recurrent dislocations, repetitive activity, pyrophosphate arthropathy, maculopathy, haemochromatosis, avascular necrosis including sickle cell disease, therapy with corticoids or other drugs, crystal deposition in cartilage including caisson disease and diseases and metabolic disorders, metaplastic or infectious arthritis (including tuberculous arthritis, meningococcal arthritis, gonococcal arthritis, salmonella arthritis), infective endocarditis, viral arthritis, recurrent arthropathy and deposition diseases of any kind such as gout, pyrophosphate arthropathy and acute calcific periarthritis.

[0288] In one embodiment of the disclosure, the inflammatory disease is a connective tissue disorder; in one embodiment, selected from the group consisting of systemic lupus erythematosus, polymyositis / dermatomyositis, systemic sclerosis, mixed connective tissue disease, sarcoidosis, and primary Sjogren's syndrome, including keratoconjunctivitis sicca, polymyalgia rheumatica, and other types of vasculitis, crystal deposition disease (including gout), pyrophosphate arthropathy, and acute calcific periarthritis.

[0289] In one embodiment of the present disclosure, the inflammatory disease is soft tissue rheumatism including bursitis, tendonitis or peritendinitis, enthesitis, nerve compression, periarthritis or capsulitis, muscle tone and muscle dysfunction.

[0290] In one embodiment of the present disclosure, the inflammatory disease is vasculitis including vasculitis secondary to rheumatoid arthritis, infectious vasculitis due to infection with bacterial species including spirochetal diseases such as Lyme disease, syphilis, rickettsial and mycobacterial infections, fungal, viral or protozoal infections, non-infectious vasculitis secondary to hypersensitivity and leukocytoplastic vasculitis including serum sickness and Henoch-Schönlein purpura, drug-induced vasculitis, essential mixed cryoglobulinemia, hypoxemia, other types of malignancies. In one embodiment, the vasculitis is selected from the group consisting of vasculitis associated with chronic tumors, non-infectious vasculitis including Takayasu's arteritis / Takayasu's disease, giant cell arteritis (temporal arteritis and polymyalgia rheumatica), Burger's disease, polyarteritis nodosa, microscopic polyarteritis, Wegener's granulomatosis, Churg-Strauss syndrome, and vasculitis secondary to connective tissue diseases including systemic lupus erythematosus, polymyositis / dermatomyositis, systemic sclerosis, mixed network tissue disease, sarcoidosis, and primary Sjogren's syndrome.

[0291] In one embodiment of the present disclosure, the inflammatory disease is an inflammatory disease of the gastrointestinal system.The inflammatory disease of the gastrointestinal system can be selected from the group consisting of inflammatory bowel disease, celiac disease, gluten-sensitive enteropathy, eosinophilic gastroenteritis, intestinal lymphectasia, inflammatory bowel disease (including Crohn's disease and ulcerative colitis), colon redundancy disease, radiation enteritis, irritable bowel syndrome, Whipple's disease, any kind of stomatitis, salivary gland disease (such as sarcoidosis, salivary duct obstruction, and Sjogren's syndrome), esophageal inflammation (for example, gastroesophageal reflux or infection with Candida species, herpes simplex and cytomegas virus), gastric inflammatory disease (including acute and chronic gastritis, Helicobacter pylori infection, Mentrier's disease), and small intestinal inflammation.

[0292] In one embodiment of the disclosure, the inflammatory disease is a neurodegenerative disease, for example, a neurodegenerative disease having an inflammatory component, for example, multiple sclerosis (MS).

[0293] In one embodiment of the present disclosure, the inflammatory disease is selected from the group consisting of dermatitis, impetigo, broid pemphigus, benign mucous membrane pemphigus, herpetic dermatitis, typical sprue, systemic amyloidosis, primary biliary cirrhosis, Gutpasture's syndrome, gout, any kind of deposition disease as pyrophosphate arthropathy and acute calcific periarthritis, pancreatitis, septic discitis, tuberculosis, malignant tumors (metastatic lesions, myeloma, etc.), spinal tumors, neoplastic spondylitis, acute disc herniation, chronic disc disease / osteoarthritis, osteoporosis, and osteomalacia, Paget's disease, hyperparathyroidism, renal osteodystrophy, spondylolysis, congenital anomaly of spinal cord ischemia, and fibromyalgia.

[0294] In one embodiment of the present disclosure, the inflammatory disease is selected from the group consisting of upper and lower respiratory tract diseases such as chronic obstructive pulmonary disease (COPD), allergic and non-allergic asthma, allergic rhinitis, allergic and non-allergic conjunctivitis, allergic and non-allergic dermatitis, and pulmonary inflammation.

[0295] Further active ingredients In some embodiments of the present disclosure, the polypeptides or polypeptide analogs disclosed herein are combined with or comprise one or more additional active ingredients, which are understood to be other therapeutic compounds or pharma- ceutically acceptable derivatives thereof.

[0296] Thus, in one embodiment, the method for treatment according to the present disclosure further comprises one or more steps of administration of one or more additional active ingredients, either simultaneously or sequentially, in any suitable ratio.

[0297] The method of treatment according to the present disclosure in one embodiment comprises the simultaneous, sequential or separate administration of a pharmaceutical composition or a polypeptide or polypeptide analogue as defined herein in combination with one or more further active ingredients.

[0298] Administration and Dosage According to the present disclosure, a composition comprising a polypeptide or polypeptide analog selected from the group consisting of SEQ ID NO:2-24, SEQ ID NO:25-47, and SEQ ID NO:48-70, or a functional variant thereof, is, in one embodiment, administered in a pharma- ceutical or therapeutically effective dose to an individual in need thereof.

[0299] A therapeutically effective amount of a polypeptide or polypeptide analog according to the present disclosure is, in one embodiment, an amount sufficient to cure, prevent, reduce the risk of, alleviate or partially arrest the clinical symptoms of a given disease or disorder and its complications. The amount effective for a particular therapeutic purpose will depend on the severity and type of disorder, as well as the weight and general condition of the subject. An amount sufficient to achieve this is defined as a "therapeutically effective amount."

[0300] In one embodiment, the composition is administered at a dose of 1 μg / day to 100 mg / day, such as 1 μg / day to 10 μg / day, such as 10 μg / day to 100 μg / day, such as 100 μg / day to 250 μg / day, such as 250 μg / day to 500 μg / day, such as 500 μg / day to 750 μg / day, such as 750 μg / day to 1 mg / day, such as 1 mg / day to 2 mg / day, such as 2 mg / day to 5 mg / day, or such as 5 mg / day to 10 mg / day, such as 10 mg / day to 20 mg / day, such as 20 mg / day to 30 mg / day, such as 30 mg / day to 40 mg / day, such as 40 mg / day to 50 mg / day, such as 50 mg / day to 75 mg / day, or such as 75 mg / day to 100 mg / day.

[0301] In one embodiment of the disclosure, a single dose of the composition is administered and may consist of 1 μg / kg body weight to 100 mg / kg body weight, such as 1-10 μg / kg body weight, such as 10-100 μg / kg body weight, such as 100-250 μg / kg body weight, such as 250-500 μg / kg body weight, such as 500-750 μg / kg body weight, such as 750 μg / kg body weight to 1 mg / kg body weight, such as 1 mg / kg body weight to 2 mg / kg body weight, such as 2-5 mg / kg body weight, such as 5-10 mg / kg body weight, such as 10-20 mg / kg body weight, such as 20-30 mg / kg body weight, such as 30-40 mg / kg body weight, such as 40-50 mg / kg body weight, such as 50-75 mg / kg body weight, or 75-100 mg / kg body weight.

[0302] In one embodiment, a dose according to the disclosure is administered once or several times per day, such as 1-6 times per day, such as 1-5 times per day, such as 1-4 times per day, such as 1-3 times per day, such as 1-2 times per day, such as 2-4 times per day, such as 2-3 times per day.

[0303] Route of administration It will be understood that the preferred route of administration will depend on the general condition and age of the subject being treated, the nature of the condition being treated, the location of the tissue being treated within the body, and the active ingredient selected.

[0304] In one embodiment of the present disclosure, the route of administration allows for introduction of the polypeptide or polypeptide analog into the bloodstream and ultimately targeting to the desired site of action.

[0305] In one embodiment of the present disclosure, the route of administration is any suitable route, such as enteral (including oral, rectal, nasal, pulmonary, buccal, sublingual, transdermal, intravesical and intraperitoneal administration), and / or parenteral (including subcutaneous, intramuscular, intrathecal, intravenous and intradermal administration). Suitable dosage forms may be prepared by conventional techniques.

[0306] Parenteral administration is any administration route other than the oral / enteral route by which the drug avoids first-pass degradation in the liver. Thus, parenteral administration includes any injection and infusion, for example, bolus injection or continuous infusion, such as intravenous, intramuscular, or subcutaneous administration. Additionally, parenteral administration includes inhalation and topical administration.

[0307] Thus, the polypeptide or polypeptide analog or composition disclosed herein, in one embodiment of the disclosure, is administered topically across any mucous membrane of the animal to which the substance or peptide is administered, for example, the mucous membrane of the nose or mouth, such as in the nose, vagina, eye, mouth, reproductive tract, lungs, gastrointestinal tract, or rectum, and thus parenteral administration can also include buccal, sublingual, nasal, rectal, vaginal and intraperitoneal administration, as well as pulmonary and bronchial administration, by inhalation or implantation. In some embodiments, the polypeptide or polypeptide analog or composition disclosed herein is administered topically across the skin.

[0308] In one embodiment of the present disclosure, intravenous, subcutaneous, and intramuscular forms of parenteral administration are used.

[0309] In one embodiment of the present disclosure, the polypeptide or polypeptide analog, or composition disclosed herein is used as a local treatment, i.e., directly introduced into the site(s) of action.Thus, the polypeptide or polypeptide analog, or composition may be applied directly to the skin or mucous membrane, or the polypeptide or polypeptide analog, or composition may be injected into the site of action, for example, into the diseased tissue or into a terminal artery that leads directly to the diseased tissue.

[0310] Pharmaceutical preparations In one embodiment of the disclosure, the polypeptide or polypeptide analog, or a pharma- ceutically acceptable derivative thereof, is administered alone or in combination with a pharma- ceutically acceptable carrier or excipient, either in single or multiple doses. The pharmaceutical composition or polypeptide according to the disclosure may be administered in combination with a pharma- ceutically acceptable carrier or diluent, as well as in the form described in Remington: The Science and Practice of Pharmacy, 20 th The compositions may be formulated with any other known adjuvants and excipients according to the prior art, such as those disclosed in the above-mentioned US Pat. No. 6,312,133, entitled "Publication Methods for the Preparation of Filtrates ...

[0311] The term "pharmaceutically acceptable derivatives" in this context includes pharmaceutically acceptable salts, which refer to salts that are not harmful to the patient. Such salts include pharmaceutically acceptable base or acid addition salts, as well as pharmaceutically acceptable metal salts, ammonium salts, and alkylated ammonium salts. Pharmaceutically acceptable derivatives further include pharmaceutically acceptable esters, prodrugs, or other precursors of a compound that can be metabolized to a biologically active compound, or crystalline forms of a compound.

[0312] Pharmaceutical or pharma- ceutically acceptable compositions may be specifically formulated for administration by any suitable route, such as enteral, oral, rectal, nasal, pulmonary, buccal, sublingual, transdermal, intravesical, intraperitoneal, and parenteral (including subcutaneous, intramuscular, intrathecal, intravenous, and intradermal) routes.

[0313] Pharmaceutical compositions for oral administration include solid dosage forms such as hard or soft capsules, tablets, troches, sugar-coated tablets, pills, lozenges, powders and granules.Optionally, they can be prepared with coatings such as enteric coatings according to methods well known in the art, or they can be formulated to provide controlled release of active ingredient, such as sustained release or sustained release.In the same solid dosage form, two active ingredients can be combined to provide controlled release of one active ingredient and immediate release of another active ingredient.

[0314] Liquid dosage forms for oral administration include solutions, emulsions, aqueous or oily suspensions, syrups and elixirs.

[0315] Pharmaceutical compositions for parenteral administration include sterile aqueous and non-aqueous injection solutions, dispersions, suspensions or emulsions, and sterile powders that are reconstituted in sterile injection solutions or dispersions before use.Depot injection preparations are also considered to be within the scope of this disclosure.Other suitable administration forms include suppositories, sprays, ointments, creams / lotions, gels, inhalants, skin patches, implants, etc.

[0316] In one embodiment of the present disclosure, the polypeptides or polypeptide analogs disclosed herein are generally utilized as free substances or as pharmaceutical derivatives, such as pharma- ceutically acceptable esters or salts thereof. Examples of the latter include acid addition salts of compounds with free base functional groups and base addition salts of compounds with free acid functional groups. The term "pharma-ceutically acceptable salts" refers to non-toxic salts of polypeptides or polypeptide analogs for use according to the present disclosure, where the salts are generally prepared by reacting a free base with a suitable organic or inorganic acid or by reacting an acid with a suitable organic or inorganic base. When a polypeptide or polypeptide analog according to the present disclosure contains a free base functional group, such salts are prepared in a conventional manner by treating a solution or suspension of the compound with a chemical equivalent of a pharma-ceutically acceptable acid. When a polypeptide or polypeptide analog according to the present disclosure contains a free acid functional group, such salts are prepared in a conventional manner by treating a solution or suspension of the compound with a chemical equivalent of a pharma-ceutically acceptable base. Physiologically acceptable salts of polypeptides or polypeptide analogs with hydroxy groups include the anionic form of the compound in combination with a suitable cation, such as sodium or ammonium ion. Other salts that are not pharmaceutically acceptable may be useful in the preparation of polypeptides or polypeptide analogs and form a further aspect of the present disclosure. Pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, trifluoroacetate, trichloroacetate, lactate, salicylate, citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoic acid) salts).

[0317] In one embodiment of the disclosure, the polypeptide or polypeptide analog disclosed herein is in a crystalline form, for example a co-crystallized form or a hydrate of a crystalline form.

[0318] The term "prodrug" refers to a peptide that is rapidly converted in vivo to the parent compound of the above formula, for example, by hydrolysis in blood or by metabolism in cells, such as cells of the basal ganglia. Thorough discussions are provided in T. Higuchi and V Stella, "Pro-drugs as Novel Delivery Systems," Vol. 14 of the ACS Symposium Series, and Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference. Examples of prodrugs include pharma- ceutically acceptable, non-toxic esters of the compounds of the present disclosure. Esters of the compounds of the present disclosure can be prepared by conventional methods, such as those described in "March's Advanced Organic Chemistry, 5 th Edition”, MB Smith & J. March, John Wiley & Sons, 2001.

[0319] In one embodiment, for parenteral administration, a solution of the polypeptide or polypeptide analog according to the present disclosure in sterile aqueous solution, aqueous propylene glycol solution, or sesame or peanut oil is used. The aqueous solution should be suitably buffered, and if necessary, the liquid diluent is made isotonic, for example, with sufficient saline or glucose. The aqueous solution is particularly suitable for intravenous, intramuscular, subcutaneous, and intraperitoneal administration. All of the sterile aqueous media used are readily available by standard techniques known to those skilled in the art.

[0320] Suitable pharmaceutical carriers include inert solid diluents or fillers, sterile aqueous solutions, and various organic solvents. Examples of solid carriers are lactose, terra alba, sucrose, cyclodextrin, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid, and lower alkyl ethers of cellulose. Examples of liquid carriers are syrup, peanut oil, olive oil, phospholipids, fatty acids, fatty acid amines, polyoxyethylene, and water. In addition, carriers or diluents can include any sustained release material known in the art, such as glyceryl monostearate or glyceryl distearate, alone or mixed with wax. The pharmaceutical composition formed by combining the compound according to the present disclosure and a pharma-ceutically acceptable carrier is then easily administered in various dosage forms suitable for the disclosed administration route. The formulations can be conveniently provided in unit dosage form by methods known in the art of pharmacy.

[0321] Formulations of the present disclosure suitable for oral administration may be presented as discrete units, such as capsules or tablets, each containing a predetermined amount of the active ingredient and which may include suitable excipients. Further, the orally available formulations may be in the form of a powder or granules, a solution or suspension in an aqueous or non-aqueous liquid, or an oil-in-water or water-in-oil liquid emulsion.

[0322] Compositions intended for oral use may be prepared according to any known method, and such compositions may contain one or more agents selected from the group consisting of sweeteners, flavoring agents, coloring agents, and preservatives to provide pharma- ceutically elegant and palatable preparations. Tablets may contain the active ingredient(s) in admixture with non-toxic pharma- ceutically acceptable excipients suitable for the manufacture of tablets. These excipients may be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrating agents, for example, corn starch or alginic acid; binding agents, for example, starch, gelatin, or acacia, and lubricants, for example, magnesium stearate, stearic acid, or talc. Tablets may be uncoated or may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over a longer period. For example, a time-delay material, such as glyceryl monostearate or glyceryl distearate, may be used.

[0323] Formulations for oral use may also be provided as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate or kaolin, or soft gelatin capsules in which the active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin, or olive oil. Aqueous suspensions may contain the compound for use according to the present disclosure in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth, and gum acacia, and dispersing or wetting agents may be naturally occurring phosphatides such as lecithin, or condensation products of alkylene oxides with fatty acids, for example, polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example, heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with fatty acids and partial esters derived from hexitols, such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with fatty acids and partial esters derived from hexitol anhydrides, for example, polyethylene sorbitan monooleate. Aqueous suspensions may also contain one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.

[0324] Oily suspensions can be formulated by suspending the active ingredient in vegetable oil, such as peanut oil, olive oil, sesame oil or coconut oil, or mineral oil, such as liquid paraffin.Oily suspensions can contain thickening agents, such as beeswax, hard paraffin or cetyl alcohol.Sweetening agents, such as those mentioned above, and flavoring agents can be added to provide a palatable oral preparation.These compositions can be preserved by the addition of antioxidants, such as ascorbic acid.

[0325] Dispersible powders and granules suitable for preparing aqueous suspension by adding water provide the active compound in a mixture with a dispersing or wetting agent, a suspending agent, and one or more preservatives.Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above.Additional excipients, such as sweeteners, flavorings, and coloring agents, can also be present.

[0326] Pharmaceutical compositions containing polypeptides or polypeptide analogs according to the present disclosure may also be in the form of oil-in-water emulsions. The oil phase may be vegetable oils, such as olive oil or peanut oil, or mineral oils, such as liquid paraffin, or mixtures thereof. Suitable emulsifiers may be naturally occurring gums, such as gum acacia or gum tragacanth, naturally occurring phosphatides, such as soybean, lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and the condensation products of said partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. Emulsions may also contain sweeteners and flavoring agents.

[0327] Syrups and elixirs may be formulated with sweetening agents, for example, glycerol, propylene glycol, sorbitol or sucrose. Such formulations may contain a demulcent, a preservative, as well as flavoring and coloring agents. The pharmaceutical compositions may be in the form of a sterile injectable aqueous or oleaginous suspension. The suspension may be formulated according to known methods using suitable dispersing or wetting agents and suspending agents as described above. The sterile injectable preparation may also be as a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be used are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils are conveniently employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed using synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid have been found to be used in the preparation of injectables.

[0328] The composition may also be in the form of a suppository for rectal administration of the compound of the present disclosure.These compositions may be prepared by mixing the polypeptide or polypeptide analog disclosed herein with a suitable non-irritating excipient that is solid at normal temperature but liquid at rectal temperature, and therefore melts in the rectum to release the drug.Such materials include, for example, cocoa butter and polyethylene glycol.

[0329] The polypeptides and polypeptide analogs of the present disclosure can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as, but not limited to, cholesterol, stearylamine, or phosphatidylcholines.

[0330] In addition, some of the polypeptides and polypeptide analogs of the present disclosure may form solvates with water or common organic solvents. Such solvates are also encompassed within the scope of the present disclosure.

[0331] Accordingly, a further embodiment of the present disclosure provides a pharmaceutical composition comprising a polypeptide or polypeptide analog disclosed herein, or a pharma- ceutically acceptable salt, solvate, or prodrug thereof, and one or more pharma- ceutically acceptable carriers, excipients, or diluents. EXAMPLES

[0332] Example 1 Functional evaluation of annexin A1 (AnxA1) N-terminal fragments: phagocytosis of monocytes method: AnxA1 N-terminal fragments were evaluated in a whole blood phagocytosis assay. Whole blood (45 uL) from human subjects was incubated with the control peptide Anxa1 2-50 (100 nM) and test peptides Compound 1 (100 nM), Compound 2 (100 nM) and Compound 3 (30 nM), or vehicle in a 96-well plate for 1 hour. 50 μl of pHrodo E. coli BioParticles Conjugate (5 μg / well) was added to the wells and the plate was incubated at 37° C., 5% CO2 for 30 minutes. 150 μl of 4% PFA was added and the plate was incubated at 4° C. for 30 minutes. 30 μL from each well of the 96-well plate was transferred to a flow cytometry tube containing 500 μL of RPMI1640 and 1 μL of Vybrant DyeCycle dye. The tubes were incubated at 37°C and 5% CO2 for 30 minutes, after which 2.5mL of RPMI1640 medium was added to each tube. Samples were acquired on an Attun NxT flow cytometer. Monocyte phagocytosis was determined as an increase in pHrodo Green fluorescence compared to control samples.

[0333] Control peptide: Control peptide 1 (AnxA1(2-50)): AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTFNPSSDVAA (SEQ ID NO: 74)

[0334] Test peptides: Compound 1 (AnxA1 2-29(V24L)): AMVSEFLKQAWFIENEEQEYVQTLKSSK (SEQ ID NO:50) Compound 2 (AnxA1 2-34(V24L)): AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGS (SEQ ID NO:55) Compound 3 (AnxA1 2-39(V24L)): AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPY (SEQ ID NO: 60)

[0335] result: The results are shown in Table 1 below and in FIG. 1. AnxA1 2-50 induces monocyte phagocytosis at a concentration of 100 nM. AnxA1 2-50 induces a nearly two-fold increase in monocyte phagocytosis compared to vehicle. Surprisingly, the shorter AnxA1 N-terminal fragments not only retained the ability to induce monocyte phagocytosis, but also retained the ability to induce phagocytosis to a higher degree than AnxA1 2-50 (2.5-3-fold increase compared to vehicle). [Table 1]

[0336] Example 2 Functional evaluation of annexin A1 (AnxA1) N-terminal fragments: chemotaxis of neutrophils method: AnxA1 N-terminal fragments were assessed in polymorphonuclear leukocytes (PMNs) isolated from buffy coats of healthy volunteers. PMNs were resuspended in migration medium (RPMI1640 (ATCC, Cat. No. 30-2001) + 0.05% BSA-FAF) to a concentration of 5.5x106 cells / mL. Receiver wells of transwell plates were filled with 200 μL migration medium containing 10 uM of compound 1, 2 or 3. Donor plates were inserted and 75 μL of cell suspension was added to the top wells (375,000 cells / well). Plates were incubated for 1 h (37°C, 5% CO2, 95% humidity). Migrated cell numbers were assessed by inserting cells from plates into transwells to stop migration, followed by addition of 200 μL of CellTiter Glo reagent to the bottom wells. Plates were incubated for 10 min in the dark (at room temperature) and 200 μL was transferred to a white 96-well plate (Lumitrac 200). Luminescence was measured using an EnVision2104 (Perkin Elmer) (exposure time 0.1 s). The chemotactic response was estimated by calculating the fold change compared to the negative control (vehicle).

[0337] Test peptides: Compound 1 (AnxA1 2-29(V24L)): AMVSEFLKQAWFIENEEQEYVQTLKSSK (SEQ ID NO:50) Compound 2 (AnxA1 2-34(V24L)): AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGS (SEQ ID NO:55) Compound 3 (AnxA1 2-39(V24L)): AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPY (SEQ ID NO: 60)

[0338] result: The results are shown in Table 2 below and in Figure 2. All three tested AnxA1 N-terminal fragments increase PMN chemotaxis compared to vehicle at a concentration of 10 μM. This effect is greatest with compound 1 (AnxA1 2-29(V24L)), which induces a 3-fold increase in chemotaxis compared to vehicle. Compound 2 (AnxA1 2-34(V24L)) and compound 3 (AnxA1 2-39(V24L)) induce a 2-fold and 1.6-fold increase in PMN chemotaxis, respectively. [Table 2]

[0339] Example 3 Pharmacological characterization of AnxA1 N-terminal fragments method: Recombinant cells co-expressing mitochondrial apoaequorin and recombinant human type 2 formyl peptide receptor (FPR2) are grown for 18 hours and then tested in antibiotic-free medium. Cells are then detached by gentle flushing with PBS-EDTA (5 mM EDTA), harvested by centrifugation, and resuspended in assay buffer (DHEM / HAM's F12 with HEPES and 0.1% protease-free BSA). After resuspension in assay buffer, cells are then incubated with coelenterazine h (Molecular Probes) at room temperature for at least 4 hours before performing dose-response testing of receptor efficacy.

[0340] 50 μl of cell suspension is then injected into 50 μl of test or control compound (solubilized in PBS / 0.5% BSA and finally diluted from 1 mM stock solution) at increasing concentrations in a 96-well plate and incubated at room temperature for 30 minutes. The resulting light emission is recorded using a Hamamatsu functional drug screening system 6000 (FDSS6000). To standardize the recorded light emission between plates and experiments, 100 μM digitonin or 20 μM ATP is added to some wells.

[0341] Agonist activity is expressed as % of the maximal activity obtained with the internal control compound. -11 ~10 -5 A range of concentrations of M are tested. Data are presented as mean values. EC50 (i.e., the concentration that induced 50% of the maximal response) is determined by best-fit analysis after log transformation using GraphPad software (version 6.0).

[0342] Control peptide Control peptide 1 (AnxA1(2-26)): Ac-Ala-Met-Val-Ser-Glu-Phe-Leu-Lys-Gln-Ala-Trp-Phe-Ile-Glu-Asn-Glu-Glu-Gln-Glu-Tyr-Val-Gln-Thr-Val-Lys-OH (SEQ ID NO: 71) Control peptide 2 (AnxA1(2-12)): Ac-Ala-Met-Val-Ser-Glu-Phe-Leu-Lys-Gln-Ala-Trp-NH2 (SEQ ID NO: 72) Control peptide 3 (W-peptide): Trp-Lys-Tyr-Met-Val-Met (SEQ ID NO: 73)

[0343] Test peptide analogs: Analog 1 (AnxA1(2-27)): Ac-Ala Met Val Ser Glu Phe Leu Lys Gln Ala Trp Phe Ile Glu Asn Glu Glu Gln Glu Tyr Val Gln Thr Val Lys Ser (SEQ ID NO: 2) Analog 2 (AnxA1(2-31)): Ac-Ala Met Val Ser Glu Phe Leu Lys Gln Ala Trp Phe Ile Glu Asn Glu Glu Gln Glu Tyr Val Gln Thr Val Lys Ser Ser Lys Gly Gly (SEQ ID NO: 6) Analog 3 (AnxA1(2-35)): Ac-Ala Met Val Ser Glu Phe Leu Lys Gln Ala Trp Phe Ile Glu Asn Glu Glu Gln Glu Tyr Val Gln Thr Val Lys Ser Ser Lys Gly Gly Pro Gly Ser Ala (SEQ ID NO: 10) Analog 4 (AnxA1(2-39)): Ac-Ala Met Val Ser Glu Phe Leu Lys Gln Ala Trp Phe Ile Glu Asn Glu Glu Gln Glu Tyr Val Gln Thr Val Lys Ser Ser Lys Gly Gly Pro Gly Ser Ala Val Ser Pro Tyr (SEQ ID NO: 14) Analog 5 (AnxA1(2-43)): Ac-Ala Met Val Ser Glu Phe Leu Lys Gln Ala Trp Phe Ile Glu Asn Glu Glu Gln Glu Tyr Val Gln Thr Val Lys Ser Ser Lys Gly Gly Pro Gly Ser Ala Val Ser Pro Tyr Pro Thr Phe Asn (SEQ ID NO: 18) Analog 6 (AnxA1(2-47)): Ac-Ala Met Val Ser Glu Phe Leu Lys Gln Ala Trp Phe Ile Glu Asn Glu Glu Gln Glu Tyr Val Gln Thr Val Lys Ser Ser Lys Gly Gly Pro Gly Ser Ala Val Ser Pro Tyr Pro Thr Phe Asn Pro Ser Ser Asp (sequence number 21) Similar7(AnxA1(2-54)): Ac-Ala Met Val Ser Glu Phe Leu Lys Gln Ala Trp Phe Ile Glu Asn Glu Glu Gln Glu Tyr Val Gln Thr Val Lys Ser Ser Lys Gly Gly Pro Gly Ser Ala Val Ser Pro Tyr Pro Thr Phe Asn Pro Ser Ser Asp Val Ala Ala Leu His Lys Ala(sequence number 24)

Claims

1. A polypeptide or polypeptide analogue comprising at least the sequence AMVSEFLKQAWFIENEEQEYVQTVKS (SEQ ID NO: 2), or a functional variant thereof, The polypeptide is AMVSEFLKQAWFIENEEQEYVQTVKS (SEQ ID NO: 2) AMVSEFLKQAWFIENEEQEYVQTVKSS (SEQ ID NO: 3) AMVSEFLKQAWFIENEEQEYVQTVKSSK (SEQ ID NO: 4) AMVSEFLKQAWFIENEEQEYVQTVKSSKG (SEQ ID NO: 5) AMVSEFLKQAWFIENEEQEYVQTVKSSKGG (SEQ ID NO: 6) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGP (SEQ ID NO: 7) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPG (SEQ ID NO: 8) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGS (SEQ ID NO: 9) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSA (SEQ ID NO: 10) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAV (SEQ ID NO: 11) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVS (SEQ ID NO: 12) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSP (SEQ ID NO: 13) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPY (SEQ ID NO: 14) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYP (SEQ ID NO: 15) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPT (SEQ ID NO: 16) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTF (SEQ ID NO: 17) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTFN (SEQ ID NO: 18) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTFNP (SEQ ID NO: 19) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTFNPS (SEQ ID NO: 20) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTFNPSSD (SEQ ID NO: 21) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTFNPSSDVAALH (SEQ ID NO: 22) AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTFNPSSDVAALHK (SEQ ID NO: 23) A polypeptide selected from the group consisting of: AMVSEFLKQAWFIENEEQEYVQTVKSSKGGPGSAVSPYPTFNPSSDVAALHKA (SEQ ID NO: 24); or a functional variant thereof, The functional variant is a polypeptide selected from the group consisting of any one of SEQ ID NOs: 2 to 24 having 1, 2, 3, 4, or 5 amino acid substitutions. A polypeptide or polypeptide analog.

2. the alanine residue at position 10 of any one of SEQ ID NOs: 2 to 24 is substituted with an amino acid residue independently selected from the group consisting of leucine, aspartic acid, methionine, glutamic acid, isoleucine and arginine; and / or the valine residue at position 21 of any one of SEQ ID NOs: 2 to 24 is substituted with an amino acid residue independently selected from the group consisting of leucine, aspartic acid, methionine, glutamic acid, isoleucine and lysine; and / or the valine residue at position 24 of any one of SEQ ID NOs: 2 to 24 is substituted with an amino acid residue independently selected from the group consisting of leucine, aspartic acid, methionine, glutamic acid, isoleucine, arginine and lysine; and / or the valine residue at position 35 of any one of SEQ ID NOs:2 to 24 is substituted with an amino acid residue independently selected from the group consisting of glycine, alanine, serine, threonine, cysteine, leucine, isoleucine, methionine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, histidine, lysine, and arginine, e.g., lysine; and / or the leucine residue at position 50 of any one of SEQ ID NOs:23 to 24 is substituted with an amino acid residue independently selected from the group consisting of glycine, alanine, serine, threonine, cysteine, valine, isoleucine, methionine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, histidine, lysine, and arginine; A polypeptide or polypeptide analog according to claim 1.

3. 2. The polypeptide or polypeptide analog of claim 1, wherein the valine residue at position 24 of any one of SEQ ID NOs: 2 to 24 is substituted with leucine.

4. 2. The polypeptide or polypeptide analog of claim 1, selected from the group consisting of: AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KS (SEQ ID NO: 25), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSS (SEQ ID NO: 26), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSK (SEQ ID NO: 27), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKG (SEQ ID NO: 28), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGG (SEQ ID NO: 29), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGP (SEQ ID NO: 30), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPG (SEQ ID NO: 31), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPGS (SEQ ID NO: 32), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPGSA (SEQ ID NO: 33), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPGGSAX ４ (SEQ ID NO: 34), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPGGSAX ４ S (SEQ ID NO: 35), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPGGSAX ４ SP (SEQ ID NO: 36), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPGGSAX ４ SPY (SEQ ID NO: 37), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPGGSAX ４ SPYP (SEQ ID NO: 38), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPGGSAX ４ SPYPT (SEQ ID NO: 39), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPGGSAX ４ SPYPTF (SEQ ID NO: 40), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPGGSAX ４ SPYPTFN (SEQ ID NO: 41), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPGGSAX ４ SPYPTFNP (SEQ ID NO: 42), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPGGSAX ４ SPYPTFNPS (SEQ ID NO: 43), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPGGSAX ４ SPYPTFNPSSD (SEQ ID NO: 44), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPGGSAX ４ SPYPTFNPSSDVAAX ５ H (SEQ ID NO: 45), AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPGGSAX ４ SPYPTFNPSSDVAAX ５ HK (SEQ ID NO: 46), and AMVSEFLKQX １ WFIENEEQEYX ２ QTX ３ KSSKGGPGGSAX ４ SPYPTFNPSSDVAAX ５ HKA (SEQ ID NO: 47), (where X １ is selected from the group consisting of alanine, leucine, aspartic acid, methionine, glutamic acid, isoleucine, and arginine; Here, X ２ is selected from the group consisting of valine, leucine, aspartic acid, methionine, glutamic acid, isoleucine, and lysine; Here, X ３ is selected from the group consisting of valine, glycine, alanine, serine, threonine, cysteine, leucine, isoleucine, methionine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, histidine, lysine, and arginine, and is preferably leucine; Here, X ４ is selected from the group consisting of valine, glycine, alanine, serine, threonine, cysteine, leucine, isoleucine, methionine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, histidine, lysine, and arginine, and is preferably lysine; and Here, X ５ is selected from the group consisting of leucine, glycine, alanine, serine, threonine, cysteine, valine, isoleucine, methionine, proline, phenylalanine, tyrosine, tryptophan, aspartic acid, glutamic acid, asparagine, glutamine, histidine, lysine, and arginine; or X １ , X ２ , X ３ , X ４ , and X ５ and functional variants thereof having one, two, three, four or five amino acid substitutions at any position other than.

5. 2. The polypeptide or polypeptide analog of claim 1, selected from the group consisting of: AMVSEFLKQAWFIENEEQEYVQTLKS (SEQ ID NO: 48), AMVSEFLKQAWFIENEEQEYVQTLKSS (SEQ ID NO: 49), AMVSEFLKQAWFIENEEQEYVQTLKSSK (SEQ ID NO: 50), AMVSEFLKQAWFIENEEQEYVQTLKSSKG (SEQ ID NO: 51), AMVSEFLKQAWFIENEEQEYVQTLKSSKGG (SEQ ID NO: 52), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGP (SEQ ID NO: 53), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPG (SEQ ID NO: 54), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGS (SEQ ID NO: 55), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSA (SEQ ID NO: 56), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAV (SEQ ID NO: 57), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVS (SEQ ID NO: 58), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSP (SEQ ID NO: 59), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPY (SEQ ID NO: 60), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYP (SEQ ID NO: 61), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPT (SEQ ID NO: 62), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPTF (SEQ ID NO: 63), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPTFN (SEQ ID NO: 64), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPTFNP (SEQ ID NO: 65), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPTFNPS (SEQ ID NO: 66), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPTFNPSSD (SEQ ID NO: 67), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPTFNPSSDVAALH (SEQ ID NO: 68), AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPTFNPSSDVAALHK (SEQ ID NO: 69), and AMVSEFLKQAWFIENEEQEYVQTLKSSKGGPGSAVSPYPTFNPSSDVAALHKA (SEQ ID NO: 70), or a functional variant thereof having 1, 2, 3, 4 or 5 amino acid substitutions.

6. 2. The polypeptide or polypeptide analog of claim 1, wherein the N-terminal amino acid residue is acetylated and / or the C-terminal amino acid residue is amidated (-NH 2 ).

7. The polypeptide or functional variant thereof is a. binds to one or more formyl peptide receptors, including formyl peptide receptor 1 (FPR1), formyl peptide receptor 2 (FPR2), and formyl peptide receptor 3 (FPR3); and / or b. Activate and / or stimulate one or more formyl peptide receptors, including formyl peptide receptor 1 (FPR1), formyl peptide receptor 2 (FPR2), and formyl peptide receptor 3 (FPR3), and / or c. Formyl peptide receptor 1 (FPR1), formyl peptide receptor 2 (FPR2), and formyl peptide receptor 2 (FPR3). are ligands and / or agonists of one or more of the formyl peptide receptors, including formyl peptide receptor 3 (FPR3); and / or d. binds to, activates, and / or is an agonist of FPR2; and / or e. activating immune cells, and / or f. activating leukocytes, such as phagocytic leukocytes, e.g., monocytes, and / or g. activating leukocytes, such as polymorphonuclear leukocytes (PMNs), e.g., neutrophils; and / or h. activating neutrophils and / or monocytes, and / or i. activating leukocyte effector functions, such as one or more of the following: induction of neutrophil chemotaxis, recruitment of neutrophil complement receptor 3 (CR3), and activation of neutrophil NADPH-oxidase; and / or j. Inducing phagocytosis in leukocytes, such as phagocytic leukocytes, e.g., monocytes; and / or k. Inducing chemotaxis in leukocytes, such as polymorphonuclear leukocytes (PMNs), e.g., neutrophils; A polypeptide or polypeptide analog according to claim 1.

8. A polypeptide conjugate comprising the polypeptide or polypeptide analog of claim 1 and one or more branched amino acid probes, the branched amino acid probe comprises a first aminoalkyl amino acid residue; the first aminoalkyl amino acid residue is optionally covalently linked to a second aminoalkyl amino acid residue, or to a second and a third aminoalkyl amino acid residue, so as to form a linear chain of two or three aminoalkyl amino acid residues; the aminoalkyl amino acid residues are individually selected from the group consisting of lysine and ornithine; the side chain of one or more of the first, second and / or third aminoalkyl amino acid residues is each modified by attaching to the side chain amino group a molecule independently selected from the group consisting of AAAq-AAA, (aa3)p-AAAq, AAAq-(aa3)p, [(aa3)-AAA]p and [AAA-(aa3)]p; wherein q is a number selected from 0, 1, 2, and 3, p is a number selected from 1, 2, and 3, AAA is an aminoalkyl amino acid residue, and (aa3) is an amino acid residue independently selected from Arg, His, Gly, and Ala; the first aminoalkyl amino acid residue is covalently attached to the N-terminus of the polypeptide, covalently attached to the C-terminus of the polypeptide, and / or covalently attached to a side chain amino group of an aminoalkyl amino acid residue within the polypeptide; However, the branched amino acid probe is composed of 2 to 9 amino acid residues. Polypeptide conjugates.

9. 9. The polypeptide conjugate of claim 8, wherein the branched amino acid probe is selected from the group consisting of: a. (Lys)Lys １ -, (Lys-Lys)Lys １ -, (Lys-Lys-Lys)Lys １ -, (Lys-Lys-Lys-Lys)Lys １ -, (Lys-Gly-Lys)Lys １ -, (Lys-Lys-Gly)Lys １ -, (Lys-Gly)Lys １ -, (Lys-Ala-Lys)Lys １ -, (Lys-Lys-Ala)Lys １ -, (Lys-Ala)Lys １ -, (Lys-His-Lys)Lys １ -, (Lys-Lys-His)Lys １ -, (Lys-His)Lys １ -, (Lys-Arg-Lys)Lys １ -, (Lys-Lys-Arg)Lys １ -, and (Lys-Arg)Lys １ -, (wherein the first lysine residue (Lys １ -) is optionally N-terminally acetylated or C-terminally amidated); b.Ac-(Ac-Lys)Lys １ -、Ac-(Ac-Lys-Lys)Lys １ -、Ac-(Ac-Lys-Lys-Lys)Lys １ -、Ac-(Ac-Lys-Lys-Lys-Lys)Lys １ -、Ac-(Ac-Lys-Gly-Lys)Lys １ -、Ac-(Ac-Lys-Lys-Gly)Lys １ -、Ac-(Ac-Lys-Gly)Lys １ -、Ac-(Ac-Lys-Ala-Lys)Lys １ -、Ac-(Ac-Lys-Lys-Ala)Lys １ -、Ac-(Ac-Lys-Ala)Lys １ -、Ac-(Ac-Lys-Hi-Lys)Lys １ -、Ac-(Ac-Lys-Lys-His)Lys １ -、Ac-(Ac-Lys-His)Lys １ -、Ac-(Ac-Lys-Arg-Lys)Lys １ -、Ac-(Ac-Lys-Lys-Arg)Lys １ -、--Ac-(Ac-Lys-Arg)Lys １ -; c. (Ac-Lys)Lys １ -NH ２ 、(Ac-Lys-Lys)Lys １ -NH ２ 、(Ac-Lys-Lys-Lys)Lys １ -NH ２ 、(Ac-Lys-Lys-Lys-Lys)Lys １ -NH ２ 、(Ac-Lys-Gly-Lys)Lys １ -NH ２ 、(Ac-Lys-Lys-Gly)Lys １ -NH ２ 、(Ac-Lys-Gly)Lys １ -NH ２ 、(Ac-Lys-Ala-Lys)Lys １ -NH ２ 、(Ac-Lys-Lys-Ala)Lys １ -NH ２ 、(Ac-Lys-Ala)Lys １ -NH ２ 、(Ac-Lys-His-Lys)Lys １ -NH ２ 、(Ac-Lys-Lys-His)Lys １ -NH ２ 、(Ac-Lys-His)Lys １ -NH ２ 、(Ac-Lys-Arg-Lys)Lys １ -NH ２ 、(Ac-Lys-Lys-Arg)Lys １ -NH ２ 、及び(Ac-Lys-Arg)Lys １ -NH ２ ; d.Ac-(Ac-Lys)Lys-Lys-、(Ac-Lys)Lys-Lys-、Ac-(Lys)Lys-Lys-、(Lys)Lys-Lys-、(Ac-Lys)Lys-Lys-NH ２ 、(Lys)Lys-Lys-NH ２ ; e.Ac-Lys-(Ac-Lys)Ly-、Lm-(Ac-Lys)Ly-、Lm-Lm-Ly- ２ 、Lys-(Lys)Lys-NH ２ ; f.Ac-(Ac-+++++)-+++、(Ac-+++++)-++、Ac-(++++)-+ ２ --Lys-Lys)-Lys-NH ２ 。

10. A pharmaceutical composition comprising the polypeptide or polypeptide analog of claim 1.

11. A drug for use in the treatment of an ischemic and / or inflammatory condition, comprising a polypeptide or polypeptide analogue according to claim 1.

12. The ischemic condition is secondary ischemia; ischemia due to stroke, injury, septic shock, systemic hypotension, cardiac arrest due to heart attack, cardiac arrhythmia, atherosclerotic disease with thrombosis, embolism of the blood vessels of the heart or any organ, vasospasm, aortic aneurysm or aneurysm of other organs, coronary artery stenosis, myocardial infarction, angina pectoris, pericarditis, myocarditis, myxedema, or endocarditis; ischemia related to major surgery; ischemia associated with solid organ transplantation; Postoperative systemic inflammatory response syndrome (SIRS), postoperative organ dysfunction, and postoperative renal failure; and reperfusion injury 12. The agent for use according to claim 11, selected from the group consisting of:

13. 12. The method of claim 11, wherein the inflammatory disease is selected from the group consisting of arthropathy (joint disease), rheumatoid arthritis (RA), gout, inflammatory diseases of the gastrointestinal system, multiple sclerosis, liver inflammation, and non-alcoholic steatohepatitis (NASH).

Images