Methods and medicaments for the treatment of respiratory inflammation and related diseases and conditions

Abstract

The present invention relates to the use of MayDay peptide fragments designated decorin 31-170, decorin 31-188, decorin 31-177, decorin 31-169, or decorin 1-188 (the N-terminal fragment of the decorin protein) in the treatment and prevention of respiratory inflammation and related disorders such as acute respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease (COPD), asthma, bronchitis, and pulmonary fibrosis. Methods for modulating tissue repair and stem cell mobilization are also contemplated in the present invention.

Description

[Technical Field]

[0001] Technical Field The present invention relates to methods of treating or preventing diseases or conditions by administration of bioactive MayDay polypeptides, and more particularly to methods of treating or preventing respiratory inflammation and / or diseases or conditions associated with respiratory inflammation by administration of MayDay polypeptides as described herein.

[0002] Background of the Invention The following contains information that may be useful in understanding the present invention. None of the information provided herein is an admission that it is prior art or relevant to the presently described or claimed invention, or that any publication or document referenced specifically or implicitly is prior art. Any discussion of prior art throughout this specification should not be taken as an admission that such prior art is, in any way, widely known or forms part of the common general knowledge in the art.

[0003] Although significant progress has been made in the treatment, and in some cases, prevention, of a significant number of diseases, many diseases and conditions remain difficult to treat or have no effective treatments available at all. In other cases, treatments may be available, but their use is accompanied by one or more undesirable side effects. For example, certain diseases and conditions involving a subject's immune system remain difficult to treat effectively because treatment modalities may have adverse effects on otherwise beneficial or important aspects of the immune response, such as the use of immunosuppressants and the associated increased susceptibility to infections.

[0004] Thus, the treatment or prevention of various diseases and conditions, such as respiratory inflammation and / or one or more diseases or conditions associated with respiratory inflammation, remains problematic.

[0005] There is a need for the development of new and improved methods of treating respiratory inflammation and / or diseases or conditions associated with respiratory inflammation, including the development of therapeutic agents suitable for use in such treatments.

[0006] The present invention seeks to provide one or more methods of treating or preventing respiratory inflammation and / or diseases or conditions associated with respiratory inflammation, for example by administration of a MayDay polypeptide as disclosed herein, and / or to at least provide a useful alternative to existing methods and / or to at least provide a useful choice for the public.

[0007] Summary of the Invention In a first aspect, the present invention relates to a method for treating or preventing respiratory inflammation and / or a disease or condition associated with respiratory inflammation in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a MayDay polypeptide as described herein.

[0008] In one example, the present invention relates to a method for treating or preventing a disease or condition associated with respiratory inflammation in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a MayDay polypeptide as described herein.

[0009] In various examples, diseases or conditions associated with respiratory inflammation include, but are not limited to, acute respiratory distress syndrome (ARDS) (including pathogen-induced ARDS, smoke-induced ARDS, pneumonia-induced ARDS, sepsis-induced ARDS, inhalation injury-induced ARDS (including ARDS associated with inhalation of water, vomit, smoke, or chemicals), immune response-induced ARDS (such as transfusion-related ARDS), and acute pancreatitis-induced ARDS); sepsis; chronic obstructive pulmonary disease; interstitial lung disease; asthma (including eosinophilic asthma, neutrophilic asthma, granulocyte-recruiting asthma, and mixed granulocyte asthma). and bronchiolitis obliterans.

[0010] In one particular example, the disease or condition is selected from the group consisting of acute respiratory distress syndrome, chronic obstructive pulmonary disease, and asthma.

[0011] In another aspect, the present invention relates to a method for treating or preventing acute respiratory distress syndrome and / or chronic obstructive pulmonary disease and / or asthma, the method comprising the step of administering to said subject a therapeutically effective amount of a MayDay polypeptide as described herein.

[0012] In various examples, the MayDay polypeptide is a) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 1 to 188 of mammalian decorin; b) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31 to 188 of mammalian decorin; c) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 1-188 of mammalian decorin; d) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 31 to 188 of mammalian decorin; e) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 1 to 177 of mammalian decorin; f) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31 to 177 of mammalian decorin; g) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 1-177 of mammalian decorin; h) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 31 to 177 of mammalian decorin; i) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 1 to 170 of mammalian decorin; j) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31 to 170 of mammalian decorin; k) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31 to 169 of human decorin; l) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 1-170 of mammalian decorin; m) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 31-170 of mammalian decorin; n) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence set forth in any one of SEQ ID NOs: 1 to 13; o) a polypeptide comprising, consisting essentially of, or consisting of at least about 10 contiguous amino acids from any one of a) through n) above; p) a polypeptide comprising or consisting of at least about 10 consecutive amino acids from any one of a) to o) above, wherein the polypeptide comprises a motif or region capable of interacting with or recruiting one or more cells, including one or more stem cells; q) a polypeptide comprising or consisting of at least about 10 consecutive amino acids from any one of a) to p) above, wherein the polypeptide comprises a motif or region capable of interacting with or recruiting one or more mesenchymal stem cells; r) a functional fragment, functional variant, peptide analog, or peptidomimetic, or derivative of any one of a) to q) above; and s) A polypeptide having at least about 90% amino acid identity to any one of a) to r) above. and wherein the polypeptide is an isolated, purified, recombinant, or synthetic polypeptide selected from the group comprising:

[0013] Any of the examples described herein may relate to any of the aspects presented herein.

[0014] In one example, the polypeptide is provided as a composition, such as a pharmaceutical composition, comprising one or more polypeptides disclosed herein together with a carrier, such as a pharmaceutically acceptable carrier. a) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 1 to 188 of mammalian decorin; b) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31 to 188 of mammalian decorin; c) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 1-188 of mammalian decorin; d) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 31 to 188 of mammalian decorin; e) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 1 to 177 of mammalian decorin; f) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31 to 177 of mammalian decorin; g) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 1-177 of mammalian decorin; h) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 31 to 177 of mammalian decorin; i) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 1 to 170 of mammalian decorin; j) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31 to 170 of mammalian decorin; k) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31 to 169 of human decorin; l) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 1-170 of mammalian decorin; m) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 31-170 of mammalian decorin; n) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence set forth in any one of SEQ ID NOs: 1 to 13; o) a polypeptide comprising, consisting essentially of, or consisting of at least about 10 contiguous amino acids from any one of a) through n) above; p) a polypeptide comprising or consisting of at least about 10 consecutive amino acids from any one of a) to o) above, wherein the polypeptide comprises a motif or region capable of interacting with or recruiting one or more cells, including one or more stem cells; and q) a polypeptide comprising or consisting of at least about 10 consecutive amino acids from any one of a) to p) above, wherein the polypeptide comprises a motif or region capable of interacting with or recruiting one or more mesenchymal stem cells; r) a functional fragment, functional variant, peptide analog, or peptidomimetic, or derivative of any one of a) to q) above; and s) a polypeptide having at least about 90% amino acid identity to any one of a) to r) above; t) Any combination of two or more of the above a) to s) The present invention is provided as a pharmaceutically acceptable composition comprising one or more polypeptides selected from the group consisting of:

[0015] In one example, the composition comprises a pharmaceutically acceptable carrier.

[0016] In a further aspect, the invention relates to a method of mediating a biological effect in a subject suffering from or susceptible to respiratory inflammation and / or a disease or condition associated with respiratory inflammation, the method comprising: a) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 1 to 188 of mammalian decorin; b) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31 to 188 of mammalian decorin; c) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 1-188 of mammalian decorin; d) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 31 to 188 of mammalian decorin; e) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 1 to 177 of mammalian decorin; f) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31 to 177 of mammalian decorin; g) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 1-177 of mammalian decorin; h) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 31 to 177 of mammalian decorin; i) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 1 to 170 of mammalian decorin; j) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31 to 170 of mammalian decorin; k) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31 to 169 of human decorin; l) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 1-170 of mammalian decorin; m) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 31-170 of mammalian decorin; n) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence set forth in any one of SEQ ID NOs: 1 to 13; o) a polypeptide comprising, consisting essentially of, or consisting of at least about 10 contiguous amino acids from any one of a) through n) above; p) a polypeptide comprising or consisting of at least about 10 consecutive amino acids from any one of a) to o) above, wherein the polypeptide comprises a motif or region capable of interacting with or recruiting one or more cells, including one or more stem cells; and q) a polypeptide comprising or consisting of at least about 10 consecutive amino acids from any one of a) to p) above, wherein the polypeptide comprises a motif or region capable of interacting with or recruiting one or more mesenchymal stem cells; r) a functional fragment, functional variant, peptide analog, or peptidomimetic, or derivative of any one of a) to q) above; and s) A polypeptide having at least about 90% amino acid identity to any one of a) to r) above. t) Any combination of two or more of the above a) to s) administering to the subject an effective amount of a polypeptide selected from the group comprising:

[0017] In one example, the biological effect is mediated in vivo by administration of one or more of the polypeptides to a subject in need thereof, for example, a subject suffering from or susceptible to respiratory inflammation and / or a disease or condition associated with respiratory inflammation. In one example, the effective amount is a therapeutically effective amount.

[0018] In another example, the biological effect is mediated ex vivo, e.g., in vitro, by contacting, e.g., a biological sample from a subject, with one or more of the polypeptides. For example, the biological effect is mediated in vitro by contacting one or more cells, one or more tissues, or one or more organs from a subject suffering from or susceptible to a disease or condition associated with respiratory inflammation with one or more of the polypeptides.

[0019] In another aspect, the present invention relates to a method of modulating tissue repair in a subject suffering from or susceptible to respiratory inflammation and / or a disease or condition associated with respiratory inflammation, the method comprising administering to the subject a therapeutically effective amount of a polypeptide as described herein.

[0020] In certain instances, a therapeutically effective amount is sufficient to recruit one or more stem cells, for example, to the site of administration or to the site where the administered polypeptide is localized.

[0021] In yet another aspect, the present invention relates to a method of modulating stem cell mobilization or related processes in a subject suffering from or susceptible to respiratory inflammation and / or a disease or condition associated with respiratory inflammation, the method comprising administering to the subject a therapeutically effective amount of a polypeptide as described herein.

[0022] In various examples, the relevant processes are angiogenesis, hematopoiesis, protein expression, induction or deposition, tissue remodeling, repair or regeneration, cell proliferation, cell differentiation (including stem cell differentiation), cell regulation, apoptosis, modulation of one or more immune responses, tumorigenesis, chemotaxis, or modulation of cell recruitment.

[0023] In various examples, the method includes administering to the subject a therapeutically effective amount of a pharmaceutically acceptable composition as described herein. In one example, administration is parenteral, e.g., intravenous, or a non-intravenous parenteral route of administration, e.g., intramuscular, intradermal, or subcutaneous injection. In another example, administration is via inhalation, instillation, or insufflation, e.g., nasal inhalation, intranasal instillation, or nasal insufflation. In one example, administration is via spray.

[0024] In another aspect, the present invention relates to a MayDay polypeptide as contemplated herein for treating respiratory inflammation and / or a disease or condition associated with respiratory inflammation in a subject in need thereof.

[0025] In another aspect, the present invention relates to a MayDay polypeptide as contemplated herein for treating acute respiratory distress syndrome and / or chronic obstructive pulmonary disease and / or asthma in a subject in need thereof.

[0026] In another aspect, the present invention relates to a MayDay polypeptide as contemplated herein for mediating a biological effect in a subject, for modulating stem cell mobilization or related processes in a subject, or for modulating tissue repair in a subject, wherein the subject is suffering from or susceptible to respiratory inflammation and / or a disease or condition associated with respiratory inflammation.

[0027] In another aspect, the invention relates to the use of a MayDay polypeptide as contemplated herein in the preparation of a medicament for the treatment of respiratory inflammation and / or a disease or condition associated with respiratory inflammation.

[0028] In another aspect, the invention relates to the use of a MayDay polypeptide as contemplated herein in the preparation of a medicament for the treatment of acute respiratory distress syndrome and / or chronic obstructive pulmonary disease and / or asthma.

[0029] In another aspect, the present invention relates to the use of a MayDay polypeptide as contemplated herein in the preparation of a medicament for mediating a biological effect in a subject, for modulating stem cell mobilization or related processes in a subject, or for modulating tissue repair in a subject, wherein the subject is suffering from or susceptible to respiratory inflammation and / or a disease or condition associated with respiratory inflammation.

[0030] In one example, the pharmaceutical agent is formulated for parenteral administration. For example, the pharmaceutical agent is formulated for intravenous administration, or for a parenteral route other than intravenous administration, such as intramuscular administration, intradermal administration, or subcutaneous administration. In another example, the pharmaceutical agent is formulated for administration by inhalation, drops, or insufflation, such as nasal inhalation, intranasal drops, or nasal insufflation. In one example, the pharmaceutical agent is formulated for administration by spray.

[0031] In various examples, the polypeptides described herein are administered to the subject at a dose of about 10 ng / kg to about 20 mg / kg. For example, the polypeptides described herein are administered at a dose of about 10 ng / kg to about 10 mg / kg, or about 50 ng / kg to about 5 mg / kg, 100 ng / kg to about 5 mg / kg, or about 1 mg / kg to about 5 mg / kg. In various examples, the polypeptides described herein are administered to the subject at a dose consistent with the doses exemplified in the Examples herein.

[0032] Other objects, aspects, features, and advantages of the present invention will become apparent from the following description. It should be understood, however, that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description. [Brief explanation of the drawings]

[0033] [Figure 1]Figure 1 provides a summary of the data presented in Example 1 herein, including data regarding the effect of administration of recombinant MayDay polypeptide (MD) on cytokine activity, myeloperoxidase (MPO) concentration, neutrophil counts, and lung tissue histology in bronchoalveolar lavage fluid samples from a mouse model of respiratory inflammation, where changes are expressed relative to untreated controls (Group 2). [Figure 2] Figure 2 shows two graphs presenting data on the effect of administration of recombinant MayDay polypeptide (MD) or dexamethasone (Dex) on tumor necrosis factor-α concentrations in bronchoalveolar lavage fluid samples from a mouse model of respiratory inflammation, collected 6 hours (Figure 2A) and 24 hours (Figure 2B) after administration, as described in Example 1 herein. Data are expressed as mean values, and error bars indicate the standard deviation for each group. Statistical significance was calculated via two-way analysis of variance compared to vehicle control: * p < 0.05; ** p < 0.01; **** p < 0.0001. [Figure 3] Figure 3 shows two graphs presenting data on the effect of administration of recombinant MayDay polypeptide (MD) or dexamethasone (Dex) on interferon-gamma concentrations in bronchoalveolar lavage fluid samples from a mouse model of respiratory inflammation, collected 6 hours (Figure 3A) and 24 hours (Figure 3B) after administration, as described in Example 1 herein. Data are expressed as mean values, and error bars indicate the standard deviation for each group. Statistical significance was calculated via two-way analysis of variance compared to vehicle control; where "*" indicates p≦0.05. [Figure 4]Figure 4 shows two graphs presenting data on the effect of administration of recombinant ayDay polypeptide (MD) or dexamethasone (Dex) on interleukin-6 concentrations in bronchoalveolar lavage fluid samples from a mouse model of respiratory inflammation, collected 6 hours (Figure 4A) and 24 hours (Figure 4B) after administration, as described in Example 1 herein. Data are expressed as mean values, and error bars indicate the standard deviation for each group. Statistical significance was calculated via two-way analysis of variance compared to vehicle control: where "**" p < 0.01; "****" p < 0.0001. [Figure 5] 5 is a graph presenting data on the effect of administration of recombinant MayDay polypeptide (MD) or dexamethasone (Dex) on neutrophil counts in bronchoalveolar lavage fluid samples from a mouse model of respiratory inflammation, collected 6, 24, and 72 hours after administration, as described in Example 1 herein, where the percentage of neutrophils (%) is normalized to the total white blood cell count. Data are expressed as mean values, and error bars indicate the standard deviation for each group. Statistical significance was calculated via two-way analysis of variance compared to vehicle control: where "**" p≦0.01; "***" p≦0.001; and "****" p≦0.0001. [Figure 6] 6 is a graph presenting data regarding the effect of administration of recombinant MayDay polypeptide (MD) or dexamethasone (Dex) on myeloperoxidase (MPO) concentrations in bronchoalveolar lavage fluid samples from a mouse model of respiratory inflammation, collected 6 and 24 hours after administration, as described in Example 1 herein. Data are expressed as mean values, and error bars indicate the standard deviation for each group. Statistical significance was calculated via two-way analysis of variance compared to vehicle control: * p < 0.05; *** p < 0.001; **** p < 0.0001. [Figure 7]Figure 7 shows three graphs presenting data on the effect of administration of recombinant MayDay polypeptide (MD) or dexamethasone (Dex) on neutrophil counts (neutrophil score), as assessed by histological analysis of tissue samples from a mouse model of respiratory inflammation taken 6 hours (Figure 7A), 24 hours (Figure 7B), and 72 hours (Figure 7C) after administration, as described in Example 1 herein. Data are expressed as mean values, and error bars indicate the standard deviation for each group. Statistical significance was calculated via two-way analysis of variance compared to vehicle control; where "*" indicates p≦0.05. [Figure 8] Figure 8 shows three graphs presenting data on the effect of administration of recombinant MayDay polypeptide (MD) or dexamethasone (Dex) on inflammation (inflammation-neutrophil alveolar score) as assessed by histological analysis of tissue samples from a mouse model of respiratory inflammation taken 6 hours (Figure 8A), 24 hours (Figure 8B), and 72 hours (Figure 8C) after administration, as described in Example 1 herein. Data are expressed as mean values, and error bars indicate the standard deviation for each group. Statistical significance was calculated via two-way analysis of variance compared to vehicle control: where "*" p≦0.05; "**" p≦0.01. [Figure 9] Figure 9 shows three graphs presenting data on the effect of administration of recombinant MayDay polypeptide (MD) or dexamethasone (Dex) on inflammatory damage (interstitial inflammation score), as assessed by histological analysis of tissue samples from a mouse model of respiratory inflammation taken 6 hours (Figure 9A), 24 hours (Figure 9B), and 72 hours (Figure 9C) after administration, as described in Example 1 herein. Data are expressed as mean values, and error bars indicate the standard deviation for each group. Statistical significance was calculated via two-way analysis of variance compared to vehicle control; where "*" indicates p≦0.05.

[0034] Detailed Description The present invention relates to a method for treating or preventing respiratory inflammation and / or a disease or condition associated with respiratory inflammation in a subject in need thereof, the method comprising the administration of a MayDay polypeptide as described herein.

[0035] Without wishing to be bound by any theory, Applicants have determined that exemplary MayDay polypeptides as contemplated herein are bioactive and capable of eliciting one or more biological responses, e.g., one or more biological responses in a subject to which they are administered. In certain instances, and again without wishing to be bound by any theory, the biological responses are useful for treating or preventing respiratory inflammation, including respiratory inflammation associated with diseases or conditions. Exemplary diseases or conditions associated with respiratory inflammation include, but are not limited to, acute respiratory distress syndrome (ARDS), asthma, sepsis, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, cystic fibrosis, and asbestosis.

[0036] In some instances, the one or more biological responses include one or more of cell activation, cell proliferation, cell chemotaxis and / or recruitment, cell differentiation, cell regulation including cell cycle regulation, and apoptosis. In particular, the one or more biological responses include one or more of stem cell activation, stem cell proliferation, stem cell chemotaxis and / or recruitment, stem cell differentiation, stem cell regulation, and stem cell apoptosis.

[0037] In some examples, the one or more biological responses include one or more of the elicitation or modulation of an immune response (including, but not limited to, upregulation or downregulation of cytokine expression, and upregulation or downregulation of immune cell (e.g., neutrophils, macrophages) activation, proliferation, maturation, or recruitment).

[0038] In some examples, the one or more biological responses include one or more of angiogenesis and / or modulation of vascularization, hematopoiesis and / or modulation of hematopoiesis, tissue remodeling and / or modulation of tissue repair, wound healing and / or regeneration, modulation of the tissue microenvironment, and modulation of tumorigenesis.

[0039] Various aspects of the invention are described in further detail in the following subsections. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification (including definitions) will control. Although methods and materials similar or equivalent to those described herein can be used in the practice of the present invention, examples of suitable methods and materials are described below. The materials, methods, and examples described herein are illustrative only and are not intended to be limiting.

[0040] Selected Definitions Reference to a range of numbers disclosed herein (e.g., 1 to 10) also includes reference to all rational numbers within that range (e.g., 1, 1.1, 2, 3, 3.9, 4, 5, 6, 6.5, 7, 8, 9, and 10) and also to any range of rational numbers within that range (e.g., 2 to 8, 1.5 to 5.5, and 3.1 to 4.7). These are merely examples of what is specifically intended, and all possible combinations of numerical values ​​from the lowest value to the highest value recited should be considered to be expressly set forth in this application as well.

[0041] Those skilled in the art will understand the meaning of the various terms of degree used herein. For example, as used herein in the context of referring to an amount (e.g., "about 9%"), the term "about" refers to an amount that is close to and includes the stated amount that still performs the desired function or achieves the desired result; for example, "about 9%" can include 9% and amounts close to 9% that perform the desired function or achieve the desired result. For example, the term "about" can refer to an amount that is less than 10%, less than 5%, less than 1%, less than 0.1%, or less than 0.01% of the stated amount. When the term "about" is used in reference to, for example, a number, concentration, amount, integer, or numerical value, it is also intended that the exact number, concentration, amount, integer, or numerical value is specifically contemplated.

[0042] As used herein, the term "and / or" may mean "and" or "or."

[0043] The terms "comprise," "comprises," and "comprising" as used in this specification and claims should not be construed in an exclusive or exhaustive sense, but rather mean "consisting of at least a portion of." When interpreting each statement in this specification that includes the term "comprise," "comprises," or "comprising," features other than those preceded by the term may also be present. Related terms such as "including," "include," and "includes" should be interpreted in the same manner.

[0044] As used herein, the term "consisting essentially of" refers to the features described and allows for the presence of other features that do not materially alter the basic characteristics of the specified features.

[0045] As used herein, the term "consisting of" means the specified materials or steps of the claimed invention, excluding any element, step, or ingredient not specified in the claim.

[0046] MayDay Polypeptide The present invention relates to the therapeutic use of one or more MayDay polypeptides. As contemplated herein, a MayDay polypeptide comprises, consists essentially of, or consists of an amino acid sequence corresponding to the amino acid sequence of an N-terminal fragment of a mammalian decorin.

[0047] Decorin (Homo sapiens gene identification number: 1634, NCBI (National Center for Biotechnology Information) reference sequence: NG_011672.2), an extracellular matrix protein, is involved in the assembly of collagen fibers and has been reported to play a role in angiogenesis and tumorigenesis.

[0048] Table 1 below provides the amino acid sequence of the full-length ovine decorin preproprotein [SEQ ID NO: 1], the amino acid sequence of the human decorin isoform A preproprotein (NCBI accession: NP_598010.1, GI: 19743846 [SEQ ID NO: 5]), the amino acid sequence of the mouse decorin preproprotein (NCBI reference sequence: NP_001177380.1 [SEQ ID NO: 8]), and the amino acid sequence of the full-length bovine decorin preproprotein [SEQ ID NO: 11]. 31~170 The amino acid sequence of the polypeptide [SEQ ID NO: 2], and the amino acid sequences of two N-terminal fragments of ovine decorin, each terminating in a cleavage site by matrix metalloproteinase 12, MayDay 31~177 [SEQ ID NO: 3] and MayDay 31~188 [SEQ ID NO: 4] are also shown in Table 1. Human MayDay 31~169 Amino acid sequence of the polypeptide [SEQ ID NO: 6], human MayDay 31~187 Polypeptide [SEQ ID NO: 7], mouse MayDay 31~164 Polypeptide [SEQ ID NO: 9], mouse MayDay 31~182 Polypeptide [SEQ ID NO: 10], bovine MayDay 31~170 Polypeptide [SEQ ID NO: 12], and bovine MayDay31~188 The polypeptide [SEQ ID NO: 13] is also presented in Table 1.

[0049] [Table 1] TIFF2026504985000003.tif233169

[0050] In one example, the MayDay polypeptide is a) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 1 to 188 of mammalian decorin; b) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31 to 188 of mammalian decorin; c) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 1-188 of mammalian decorin; d) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 31 to 188 of mammalian decorin; e) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 1 to 177 of mammalian decorin; f) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31 to 177 of mammalian decorin; g) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 1-177 of mammalian decorin; h) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 31 to 177 of mammalian decorin; i) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 1 to 170 of mammalian decorin; j) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31 to 170 of mammalian decorin; k) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31 to 169 of human decorin; l) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 1-170 of mammalian decorin; m) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 31-170 of mammalian decorin; n) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence set forth in any one of SEQ ID NOs: 1 to 13; o) a polypeptide comprising, consisting essentially of, or consisting of at least about 10 contiguous amino acids from any one of a) through n) above; p) a polypeptide comprising or consisting of at least about 10 consecutive amino acids from any one of a) to o) above, wherein the polypeptide comprises a motif or region capable of interacting with or recruiting one or more cells, including one or more stem cells; and q) a polypeptide comprising or consisting of at least about 10 consecutive amino acids from any one of a) to p) above, wherein the polypeptide comprises a motif or region capable of interacting with or recruiting one or more mesenchymal stem cells; r) a functional fragment, functional variant, peptide analog, or peptidomimetic, or derivative of any one of a) to q) above; and s) A polypeptide having at least about 90% amino acid identity to any one of a) to r) above. The polypeptide is selected from the group comprising:

[0051] Those skilled in the art will recognize that, upon reading this description, they can determine that these polypeptides are representative of the MayDay polypeptides contemplated herein. A variety of therapeutic and prophylactic uses of and for these polypeptides are contemplated, for example, in therapies including cell chemotaxis and recruitment, modulation of tissue remodeling, tissue repair and wound healing, modulation of the immune response, modulation of angiogenesis, and modulation of the tissue microenvironment.

[0052] MayDay polypeptides suitable for use herein include variants or derivatives thereof, including polypeptides having an amino acid sequence identical to that found in a naturally occurring decorin protein, for example, the amino acid sequence of an N-terminal fragment of a naturally occurring decorin protein, and polypeptides having one or more amino acid mutations derived from the amino acid sequence of a naturally occurring decorin protein.

[0053] The term "amino acid" refers to natural amino acids, unnatural amino acids, and amino acid analogs. Unless otherwise specified, the term "amino acid" includes both D and L stereoisomers, if the respective structures allow for such stereoisomeric forms.

[0054] Naturally occurring amino acids include alanine (Ala or A), arginine (Arg or R), asparagine (Asn or N), aspartic acid (Asp or D), cysteine ​​(Cys or C), glutamine (Gin or Q), glutamic acid (Glu or E), glycine (Gly or G), histidine (His or H), isoleucine (He or I), leucine (Leu or L), lysine (Lys or K), methionine (Met or M), phenylalanine (Phe or F), proline (Pro or P), serine (Ser or S), threonine (Thr or T), tryptophan (Tip or W), tyrosine (Tyr or Y), and valine (Val or V).

[0055] Unnatural amino acids include azetidine carboxylic acid, 2-aminoadipic acid, 3-aminoadipic acid, β-alanine, naphthylalanine ("naph"), aminopropionic acid, 2-aminobutyric acid, 4-aminobutyric acid, 6-aminocaproic acid, 2-aminoheptanoic acid, 2-aminoisobutyric acid, 3-aminoisobutyric acid, 2-aminopimelic acid, tert-butylglycine ("tBuG"), 2,4-diaminoisobutyric acid, desmosine, 2,2'-diaminopimelic acid, 2,3-diaminopropionic acid, N-ethylglycine, N-ethylasparagine, homoproline ("hPro" or "homoP"), hydroxylysine, allo-hydroxylysine, 3-hydroxyproline ("3Hyp"), and 4-hydroxyproline. ("4Hyp"), isodesmosine, allo-isoleucine, N-methylalanine ("MeAla" or "Nime"), N-alkylglycines ("NAG"), such as N-methylglycine, N-methylisoleucine, N-alkylpentylglycines ("NAPG"), such as N-methylpentylglycine, N-methylvaline, naphthylalanine, norvaline ("Norval"), norleucine ("Norleu"), octylglycine ("OctG"), ornithine ("Orn"), pentylglycine ("pG" or "PGly"), pipecolic acid, thioproline ("ThioP" or "tPro"), homolysine ("hLys"), and homoarginine ("hArg").

[0056] The term "amino acid analog" refers to a natural or unnatural amino acid in which one or more of the C-terminal carboxyl group, the N-terminal amino group, and the side chain functional groups have been reversibly or irreversibly chemically blocked or otherwise modified to another functional group. For example, aspartic acid-(β-methyl ester) is an amino acid analog of aspartic acid; N-ethylglycine is an amino acid analog of glycine; or alanine carboxamide is an amino acid analog of alanine. Other amino acid analogs include methionine sulfoxide, methionine sulfone, S-(carboxymethyl)-cysteine, S-(carboxymethyl)-cysteine ​​sulfoxide, and S-(carboxymethyl)-cysteine ​​sulfone.

[0057] The term "expression construct" refers to a genetic construct that contains elements that allow for the transcription of a polynucleotide molecule of interest and, optionally, the translation of the transcript into a polypeptide. Expression constructs typically contain, in a 5' to 3' direction: (1) a promoter that is functional in the host cell into which the construct will be introduced; (2) a polynucleotide to be expressed, and (3) a transcription terminator that is functional in the host cell into which the construct will be introduced; Includes.

[0058] The expression constructs of the invention are inserted into replicable vectors for cloning or expression, or are integrated into a host genome.

[0059] As used herein, the term "vector" refers to a polynucleotide molecule, typically but not limited to double-stranded DNA, that is amenable to use in molecular biology techniques, for example, to modify, manipulate, replicate, amplify, or transport polynucleotide molecules. In some instances, vectors are used to transport polynucleotide molecules, such as but not limited to, gene constructs, e.g., expression constructs, into host cells or organisms. In some instances, the vectors can replicate and / or be maintained in more than one host system.

[0060] A "fragment" of a polypeptide is a subsequence of the polypeptide, typically a sequence that performs a function required for an activity, such as an enzymatic or binding activity, and / or provides the three-dimensional structure of the polypeptide or a portion thereof, e.g., an epitope. It will be understood that a fragment of a polypeptide may have or elicit a different function or set of functions than that possessed or exhibited by the full-length polypeptide from which it is derived.

[0061] As used herein, the term "peptide" refers to short polymers of amino acids linked together by peptide bonds. While it will be recognized that the names associated with various classes of amino acid polymers (e.g., peptide, protein, polypeptide, etc.) are somewhat arbitrary, peptides are generally about 50 amino acids in length or less. Peptides can contain natural amino acids, unnatural amino acids, amino acid analogs, and / or modified amino acids. Peptides can be subsequences of naturally occurring proteins or unnatural (including synthetic) sequences.

[0062] As used herein, the term "synthetic peptide" encompasses peptides having amino acid sequences that differ significantly from those found in naturally occurring peptides and / or proteins. As used herein, a "synthetic peptide" can be produced or synthesized by any suitable method (e.g., recombinant expression, e.g., via an expression construct, chemical synthesis, enzymatic synthesis, etc.), and can include any chemical modification to the parent peptide, including, but not limited to, methods such as truncation, deletion, cyclization, or synthetic or semi-synthetic non-peptidic derivatives that retain the same biological function(s) as the starting peptide. Protein synthesis methods, such as solid-phase synthesis, are well known in the art.

[0063] The terms "peptide analog," "peptide mimetic," or "peptidomimetic" refer to a peptide-like molecule that mimics a sequence derived from a protein or peptide. A peptide analog, peptide mimetic, or peptidomimetic can have amino acid and / or non-amino acid components. Examples include chemically modified peptides, peptoids (side chains attached to nitrogen atoms rather than the α-carbon of the peptide backbone), β-peptides (amino groups attached to the β-carbon rather than the α-carbon), and the like. A chemical modification includes one or more modifications at an amino acid side group, the α-carbon atom, the terminal amine group, or the terminal carboxy group. A chemical modification can be the addition of chemical moieties, the creation of new bonds, or the removal of chemical moieties. Modifications of amino acid side chains include, but are not limited to, acylation of the lysine epsilon-amino group, N-alkylation of arginine, histidine, or lysine, alkylation of the glutamic acid or aspartic acid carboxylic acid group, lactam formation via cyclization of the lysine epsilon-amino group with the glutamic acid or aspartic acid side chain carboxyl group, hydrocarbon "stapling" (e.g., to stabilize an α-helical conformation), and deamidation of glutamine or asparagine. Modifications of terminal amine groups include, but are not limited to, desamino, N-lower alkyl, N-di-lower alkyl, constrained alkyl, (e.g., branched, cyclic, fused, adamantyl), and N-acyl modifications. Modifications of terminal carboxy groups include, but are not limited to, amide, lower alkyl amide, constrained alkyl, (e.g., branched, cyclic, fused, adamantyl) alkyl, dialkyl amide, and lower alkyl ester modifications. Lower alkyl is a C1-C4 alkyl. Additionally, one or more side groups or terminal groups may be protected by protecting groups known to the ordinarily skilled peptide chemist. The α-carbon of an amino acid may be mono- or di-methylated.

[0064] It will be understood that any one of the proteins or peptides described herein in certain instances will comprise one or more unnatural amino acids, one or more amino acid analogs, or will be or comprise a synthetic peptide, synthetic polypeptide, peptide analog, or peptide mimetic. Similarly, it will be understood that any one of the proteins or peptides described herein in certain instances will be the starting point for one or more modifications, synthetic methods, or protein engineering methods to develop a peptide analog having a desired biological activity (e.g., a biological activity qualitatively similar to that of the parent protein or peptide, but of a quantitatively different magnitude, or an actual biological activity different from that elicited by the parent protein or peptide).

[0065] As used herein, the term "fused polypeptide" refers to a polypeptide comprising two or more amino acid sequences, e.g., two or more polypeptide domains, fused through their respective amino and carboxyl residues by peptide bonds to form a single continuous polypeptide. It should be understood that the two or more amino acid sequences may be directly fused or indirectly fused through their respective amino and carboxyl termini via a linker or spacer or additional polypeptide.

[0066] As used herein, the term "polypeptide" encompasses an amino acid chain of any length, but preferably at least 10 amino acids, including full-length proteins, in which the amino acid residues are linked by covalent peptide bonds. The polypeptides described herein may be purified natural products or may be produced in part or in whole using recombinant or synthetic techniques. The term may refer to a polypeptide, an aggregate of a polypeptide, such as a dimer or other multimer, a fused polypeptide, a polypeptide variant, or a derivative thereof.

[0067] It will be understood that for the specific polypeptides and proteins contemplated herein, natural variations may exist between individual organisms, including organisms of the same species or strain. These variations may be demonstrated by differences in (single) amino acid(s) in the overall sequence, or by deletions, substitutions, insertions, inversions, or additions of (single) amino acid(s) in the sequence. Amino acid substitutions that do not substantially alter biological and immunological activity are well known. Amino acid substitutions between related amino acids, or substitutions that occur frequently during evolution, include, among others, Ser / Ala, Ser / Gly, Asp / Gly, Asp / Asn, Ile / Val. Other amino acid substitutions include Asp / Glu, Thr / Ser, Ala / Gly, Ala / Thr, Ser / Asn, Ala / Val, Thr / Phe, Ala / Pro, Lys / Arg, Leu / Ile, Leu / Val, and Ala / Glu. Based on this information, methods have been developed for rapid and sensitive protein comparison and for determining functional similarity between homologous proteins. Mutations having such amino acid substitutions, as well as deletions and / or insertions, of the illustrative examples described herein are within the scope of the present invention, so long as the resulting proteins retain their activity, such as their immunoreactivity. This explains why one or more proteins described herein, when isolated from different field isolates, may have a degree of identity less than 100%, yet still represent the same protein with the same functional (e.g., immunological) characteristics. Such mutations in the amino acid sequence of a particular protein described herein still provide a protein capable of mediating one or more functions involving reacting with antibodies specific for the protein specifically identified herein, which is deemed to be an equivalent of the immunologically functional protein identified herein and, therefore, does not substantially affect the immunological functionality of the protein.

[0068] When a protein is to be used, for example, for diagnostic or therapeutic purposes, e.g., for reaction with antibodies, or to mediate a biological effect in vivo, e.g., one or more biological functions associated with the native protein or a fragment thereof, it may be convenient, but is not necessary, to use the entire protein. It is also possible to use a polypeptide fragment of the protein (e.g., intact, bound to a carrier, or as a component in a condensed polypeptide), or a polypeptide fragment or related amino acid sequence derived from the protein that is capable of the desired biological effect, e.g., eliciting an immune response against the protein, being recognized by antibodies specific for the protein, or mediating a cell signaling effect. Such a polypeptide fragment may be referred to by reference to a function it possesses, e.g., a function it shares with the full-length protein from which it is derived. For example, a polypeptide fragment that has an immunological effect may be referred to as an immunogenic fragment, where an "immunogenic fragment" is understood to be a fragment of a full-length protein that retains its ability to elicit an immune response in a vertebrate host or be recognized by antibodies specific for the parent protein. Similarly, a polypeptide fragment that retains or has one or more biological effects elicited by the full-length protein from which it is derived, or that has a related or different biological effect, is referred to herein as a "bioactive fragment" or "bioactive polypeptide fragment." Similarly, a polypeptide that has a biological effect, e.g., a polypeptide that is capable of stimulating a biological response in a cell or eliciting a therapeutic effect, may be referred to herein as a "bioactive fragment" or "bioactive polypeptide fragment" or grammatical equivalents thereof.

[0069] A wide variety of techniques are available for identifying such polypeptide fragments, as well as DNA fragments encoding such fragments. For example, in the case of immunogenic fragments, such fragments may contain one or more determinants or epitopes. Well-established empirical and computer simulation methods for epitope detection exist and are well known to those skilled in the art. For example, computer algorithms can select specific protein fragments as being immunologically important epitopes based on their sequence and / or structural identity with known epitopes. The determination of these regions is typically based on a combination of hydrophilicity criteria and secondary structural features. Immunogenic fragments (or epitopes) usually have a minimum length of 6, more commonly 8 amino acids, or more than 8, e.g., 9, 10, 12, 15, or even 20 or more amino acids. Thus, nucleic acid sequences encoding such fragments have a length of at least 18, more commonly 24, and preferably 27, 30, 36, 45, or even 60 nucleic acids.

[0070] Similarly, one of skill in the art will know how to identify bioactive fragments using a variety of assays aimed at identifying or detecting particular biological responses. Exemplary methods suitable for use in identifying or detecting bioactive fragments contemplated herein are provided below (including in the Examples).

[0071] The term "variant" when referring to a polypeptide includes naturally occurring polypeptides, recombinant polypeptides, and synthetically produced polypeptides, including those containing one or more non-naturally occurring amino acids, one or more amino acid analogs, peptide analogs, and peptide mimetics. Variant polypeptide sequences preferably have a similar or similar structure to a sequence of the invention, and more preferably at least 50%, more preferably at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 100%, at least 101%, at least 102%, at least 103%, at least 104%, at least 105%, at least 106%, at least 107%, at least 108%, at least 109%, at least 110%, at least 111%, at least 112%, at least 113%, at least 114%, at least 115%, at least 116%, at least 117%, at least The amino acid sequence of the polypeptide of the present invention may be 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity. Identity is found over a comparison window of at least 20 amino acid positions, preferably at least 50 amino acid positions, at least 100 amino acid positions, or over the full length of a polypeptide of the present invention.

[0072] Polypeptide sequence identity can be determined in the following manner: A subject polypeptide sequence is compared to a candidate polypeptide sequence using BLASTP (from the BLAST suite of programs, version 2.2.10 [October 2004]) in bl2seq, which is publicly available from NCBI (ftp: / / ftp.ncbi.nih.gov / blast / ). The default parameters of bl2seq are used, except that filtering of low-complexity regions should be turned off.

[0073] Polypeptide sequence identity can also be calculated over the entire length of overlap between a candidate polynucleotide sequence and a subject polynucleotide sequence using a global sequence alignment program. EMBOSS-needle (available at http: / / www.ebi.ac.uk / emboss / align / ), as discussed above, and GAP (Huang, X. (1994) On Global Sequence Alignment. Computer Applications in the Biosciences 10, 227-235) are also suitable global sequence alignment programs for calculating polypeptide sequence identity.

[0074] Polypeptide variants contemplated herein include those that exhibit similarity to one or more of the specifically identified sequences, likely to preserve functional equivalence of the sequences, and that would not reasonably be expected to have occurred by chance. Such sequence similarity percentages for polypeptides can be determined using the publicly available bl2seq program from the BLAST suite of programs (version 2.2.10 [October 2004]) from NCBI (ftp: / / ftp.ncbi.nih.gov / blast / ). Polypeptide sequence similarity percentages can be determined using the following unix command line parameters: bl2seq -i peptideseq1 -j peptideseq2 -FF -p blastp.

[0075] The variant polypeptide sequence preferably has a nucleotide sequence of 1×10 or less when compared to any one of the specifically identified sequences. -10 less than 1×10 -20 Less than 1×10 -30 Less than 1×10 -40 Less than 1×10 -50 Less than 1×10 -60 Less than 1×10 -70 Less than 1×10 -80 Less than 1×10 -90 Less than 1×10 -100Less than 1×10 -110 Less than 1×10 -120 Less than or 1 x 10 -123 indicates an E value less than

[0076] The parameter FF turns off filtering of low complexity segments. The parameter p selects the algorithm appropriate for pairs of sequences. The program finds regions of similarity between sequences and, for each such region, reports an "E value," which is the expected number of times one can expect to see such a match by chance in a database of a given reference size containing random sequences. For small E values, much less than 1, this is approximately the probability of such a random match.

[0077] Conservative substitutions of one or several amino acids in the described polypeptide sequences that do not significantly alter its biological activity are also included in the present invention. Those skilled in the art will know how to make phenotypically silent amino acid substitutions (see, for example, Bowie et al., 1990, Science 247, 1306).

[0078] MayDay polypeptide variants contemplated herein also include those that are produced from nucleic acid encoding the polypeptide but that differ from the wild-type polypeptide in that they have been differentially processed to have an altered amino acid sequence, e.g., in one example, a MayDay variant is produced by alternative splicing patterns of the primary RNA transcript from which the wild-type polypeptide is produced.

[0079] Therapeutic Methods and Compositions Therapies relying on the MayDay polypeptides described herein are particularly contemplated for the treatment or prevention of respiratory inflammation, eg, respiratory inflammation associated with a disease or condition, eg, in a subject in need thereof.

[0080] As used herein, a disease or condition includes a disease, disorder, pathology or condition in a subject and includes any symptoms of, or sequelae associated with, the disease, condition, disorder or pathology.

[0081] As used herein, a "subject" is an animal, typically a mammal (including companion mammals) or a human. Exemplary companion animals include cats, horses, and dogs. Exemplary farm animals include cattle, sheep, goats, deer, and pigs.

[0082] It will be understood that the various treatments contemplated herein will typically embody the administration of an effective amount of a MayDay polypeptide.

[0083] An "effective amount" is an amount sufficient to elicit one or more beneficial or desired results, including one or more clinical results. An effective amount can be administered in one or more doses by various routes of administration. The effective amount will vary depending on, among other factors, the disease or condition presented, the severity of the disease or condition, the age and relative health of the subject, the potency of the agent administered, the mode of administration, and the treatment desired. One of ordinary skill in the art will be able to determine the appropriate dose, taking these or any other relevant factors into consideration.

[0084] Contemplated are treatment and / or prevention of respiratory inflammation, such as that associated with certain diseases or conditions (including disorders, pathologies, or symptoms or sequelae thereof, such as those discussed above). Respiratory inflammation is a condition that can manifest in acute or chronic forms and has a variety of symptoms, including fatigue, dyspnea, wheezing, discomfort, and chest pain. Furthermore, respiratory inflammation is associated with many diseases or conditions that have significant clinical consequences.

[0085] Particularly contemplated examples of diseases or conditions involving respiratory inflammation include acute respiratory distress syndrome (ARDS) (including, but not limited to, pathogen-induced ARDS, smoke-induced ARDS, pneumonia-induced ARDS, sepsis-induced ARDS, inhalation injury-induced ARDS (including ARDS associated with inhalation of water, vomit, smoke, or chemicals), immune response-induced ARDS (such as transfusion-related ARDS), and acute pancreatitis-induced ARDS); sepsis; chronic obstructive pulmonary disease; interstitial lung disease; asthma (eosinophilic asthma, neutrophilic asthma, granulocyte-recruiting asthma, and mixed granulocyte-recruiting asthma). granulocytic asthma); pulmonary fibrosis; asbestosis; interstitial pneumonia; pneumonitis (including, but not limited to, lymphocytic interstitial pneumonia and hypersensitivity pneumonitis); collagen vascular diseases; granulomatous vasculitis; sarcoidosis; cystic fibrosis; drug-induced pulmonary diseases (including, but not limited to, toxin-induced pulmonary diseases (such as those associated with methotrexate, bleomycin, etc.) and immune-mediated pulmonary diseases (such as those associated with pembrolizumab)); bronchiectasis; bronchitis; bronchiolitis; and bronchiolitis obliterans.

[0086] In some instances, the disease or condition is or is associated with ARDS, such as an ARDS selected from the group consisting of pathogen-induced ARDS, smoke-induced ARDS, pneumonia-induced ARDS, sepsis-induced ARDS, inhalation injury-induced ARDS, immune response-induced ARDS, and acute pancreatitis-induced ARDS.

[0087] As used herein, the term "treatment" and related terms such as "treating" and "treat" generally refer to the treatment of a human or non-human subject in which some desired therapeutic effect is achieved. A therapeutic effect can be, for example, suppressing, reducing, ameliorating, arresting, curing, reducing, delaying the onset or slowing the progression, or preventing a disease or condition in a subject, or any activity that otherwise affects the structure or any function of the subject's body, and includes treatment that relieves, reduces, or alleviates at least one symptom or sequela in a subject, or causes a delay in the onset or slowing the progression of a disease or condition or its symptoms or its sequelae. For example, treatment can be the reduction of one or some symptoms or sequelae of a disorder, or the complete eradication of a disorder. Within the meaning of the present disclosure, the term "treat" also refers to arresting, delaying the onset (i.e., the period before clinical symptoms of a disease become apparent), and / or reducing the risk of developing or worsening a disease. "Treatment" can also mean extending survival compared to expected survival in the absence of treatment, e.g., increasing overall survival (OS) compared to a subject not receiving a treatment as described herein, and / or increasing progression-free survival (PFS) compared to a subject not receiving a treatment as described herein. The term "treating" can also mean improving the condition of a subject with a disease or condition, e.g., a reduction in one or more markers for the disease or condition in the subject, a reduction or substantial increase in the rate of disease progression in the subject, and an improvement in one or more physiological or metabolic responses or measurements in the subject (compared to one or more measurements in a subject with a similar disease or condition who is receiving no treatment or a different treatment, or compared to one or more measurements in the same subject prior to treatment). Prophylactic treatment (including treatment administered before one or more signs or symptoms of a disease or condition are manifest) is also contemplated.

[0088] As used herein, the term "stem cell" refers to a cell that is capable of self-renewal without differentiation and the ability to differentiate into other cell types. The term "stem cell," as the context requires, encompasses totipotent stem cells, pluripotent stem cells, and multipotent stem cells.

[0089] Generally, without wishing to be bound by any theory, the methods contemplated herein are, in certain instances, directed to the modulation of one or more endogenous stem cells in a subject.

[0090] Nevertheless, in certain instances, ex vivo and / or a combination of ex vivo and in vivo treatment methods are contemplated, for example, when one or more stem cells are used that have been cultured in vitro after isolation from a stem cell-containing tissue source. In one example, the stem cells are mesenchymal stem cells.

[0091] Stem cells can differentiate into other cell types and also have the ability to self-renew without differentiation. By providing a variety of differentiated functional cells as needed, stem cells are important for the formation of new tissues and also for the repair of damaged or diseased tissues [Li, L., & Xie, T. (2005). Stem cell niche: structure and function. Annu. Rev. Cell Dev. Biol., 21, 605-631.] In addition to their progenitor function, stem cells also exhibit their own functions in promoting tissue regeneration and repair, for example, through the secretion of bioactive factors and through modulation of the behavior of other cell types [Duscher, D., Barrera, J., Wong, VW, Maan, ZN, Whittam, AJ, Januszyk, M., & Gurtner, GC (2016). Stem cells in wound healing: the future of regenerative medicine? A mini-review. Gerontology, 62(2), 216-225]. As part of normal tissue growth and repair, local endogenous stem cells perform these functions; however, in the setting of extensive tissue injury or when comorbid disease factors are present, the normal endogenous stem cell population may be insufficient [Kanji, S., & Das, H. (2017). Advances of stem cell therapeutics in cutaneous wound healing and regeneration. Mediators of inflammation, 2017].

[0092] Stem cell therapy has been recognized as having great potential in regenerative medicine, and the administration of additional stem cells has been shown to be effective in a range of injuries and disease states, such as skin wounds [Falanga, V., Iwamoto, S., Chartier, M., Yufit, T., Butmarc, J., Kouttab, N., & Carson, P. (2007). Autologous bone marrow-derived cultured mesenchymal stem cells delivered in a fibrin spray accelerate healing in murine and human cutaneous wounds. Tissue engineering, 13(6), 1299-1312], nervous system injuries [di Summa, PG, Kingham, PJ, Raffoul, W., Wiberg, M., Terenghi, G., & Kalbermatten, DF (2010). Adipose-derived stem cells enhance peripheral nerve regeneration. Journal of Plastic, Reconstructive & Aesthetic Surgery, 63(9), 1544-1552] and myocardial infarction [Berry, MF, Engler, AJ, Woo, YJ, Pirolli, TJ, Bish, LT, Jayasankar, V., & Sweeney, HL (2006). Mesenchymal stem cell injection after myocardial infarction improves myocardial compliance. American Journal of Physiology-Heart and Circulatory Physiology, 290(6), H2196-H2203]. Bone marrow-derived stem cells, such as mesenchymal stem cells, can also differentiate into cells required for tissue repair at the site of injury, including pericytes, endothelial cells, and keratinocytes.Mesenchymal stem cells, along with fibroblasts, fibrocytes, and pericytes, can differentiate into myofibroblast precursor cells, which, stimulated by cell-matrix interactions, matrix stiffness, and mechanical stress, become myofibroblasts, the main matrix-producing cells in wound healing. Mesenchymal stem cells also support hematopoiesis by maintaining the microenvironment for hematopoietic stem cells. For example, in adult bone marrow, mesenchymal stem cells (MSCs) are a pool of regenerative cells that can self-renew and play an important role in tissue repair. Mesenchymal stem cells release cytokines that modulate inflammatory responses and neurotrophic factors that promote healing processes such as cell recruitment, angiogenesis, and collagen synthesis.

[0093] Mesenchymal stem cells are not only capable of differentiating into the cell types required at the site of injury, but also play an important role in regulating other cells, including immune cells and other wound-healing cells. They recruit keratinocytes, dermal fibroblasts, and other nearby stem cells by releasing paracrine signals. Mesenchymal stem cells enhance the rates of endothelial cell tube formation as well as fibroblast migration, proliferation, and collagen synthesis. Mesenchymal stem cells also secrete important wound-healing growth factors, such as keratinocyte growth factor (KGF), vascular endothelial growth factor, and platelet-derived growth factor. Finally, mesenchymal stem cells regulate matrix metalloproteinase expression and collagen deposition during remodeling.

[0094] Without wishing to be bound by any theory, Applicants believe that administration of MayDay polypeptides is effective in promoting tissue repair or regeneration, e.g., lung tissue repair or regeneration, and / or in preventing tissue injury, e.g., preventing lung tissue injury; and It is believed to be effective in treating or preventing respiratory inflammation, whether through its activity in modulating and / or mobilizing one or more stem cells, through promoting tissue repair or preventing injury, or through modulation of one or more other biological processes, such as the immune response. Such biological processes may include, but are not limited to, angiogenesis, vasculogenesis, tissue remodeling, and the enrichment and / or mobilization and / or activation of one or more cell populations, such as circulating stem cells or immune cells.

[0095] During the inflammatory stage of tissue healing, mesenchymal stem cells regulate the immune response by blocking T cell proliferation, reducing inflammatory markers such as tumor necrosis factor-α, interleukin-1 (IL-1), and IL-6, and producing anti-inflammatory markers such as IL-10 and IL-4. The immunomodulatory and anti-inflammatory activities of mesenchymal stem cells have been shown to be beneficial in the virulent respiratory inflammation associated with various diseases and conditions. Respiratory inflammation is typically associated with a significant release of dysregulated inflammatory cytokines, which leads to subsequent lung injury. Mesenchymal stem cells have been shown to exert broad anti-inflammatory effects in numerous cases of virulent respiratory inflammation, including, but not limited to, ARDS and interstitial lung disease. In particular, systemic administration of exogenous mesenchymal stem cells has been shown to improve clinical outcomes and reduce mortality in patients with moderate to severe ARDS. Without wishing to be bound by any theory, this effect is likely due to the immunomodulatory properties of mesenchymal stem cells, which block T cell proliferation, downregulate inflammatory markers, and increase anti-inflammatory markers.

[0096] It will be appreciated by those skilled in the art that various immune cell populations (e.g., macrophages, neutrophils, beta cells, and dendritic cells) may be involved in some immune responses (including inflammatory responses, such as those observed in diseases or conditions associated with respiratory inflammation). In some such diseases, other cell types, such as endothelial cells or stem cells (e.g., mesenchymal stem and progenitor cells), are expected to play a role, without wishing to be bound by any theory, in the onset, progression, and possibly resolution of diseases or conditions associated with respiratory inflammation.

[0097] Despite advances in cell administration technology, ex vivo methods can be inefficient and invasive, requiring the collection, extraction, enrichment, and re-administration of isolated cell populations of donor tissue. Therapeutic agents that can elicit one or more therapeutic effects, such as a biological response, for example, that can induce local mobilization of endogenous stem cells, could be of considerable use in treating conditions that may benefit from stem cell activity.

[0098] Stem cell therapies, such as those utilizing mesenchymal stem cells, may, in some instances, involve the extraction of a stem cell population from a donor, followed by ex vivo culture and eventual administration. Stem cells may be derived from either autologous or allogeneic sources. Autologous mesenchymal stem cells are derived from the subject to whom the treatment is to be administered, effectively with the subject acting as their own donor. Allogeneic mesenchymal stem cells are derived from a human other than the patient, either an HLA-matched related or unrelated donor. Therapies based on the administration of either allogeneic or autologous mesenchymal stem cells require extraction from the donor prior to ex vivo culture and eventual administration. Such therapies may sometimes require ongoing concurrent therapy, such as immunosuppression. These cell-based therapies are costly, complex, challenging from a quality control perspective, and may require ongoing patient monitoring.

[0099] In contrast, exemplary methods contemplated herein involve the use of endogenous cells (such as endogenous mesenchymal stem cells), which are cells that are already present in a subject or can be induced in a subject. These therapies therefore avoid the need for cell extraction or culture, and do not require any immunosuppression, as endogenous cells are immunologically "autologous." Thus, pharmacological activation of endogenous stem cells, such as activation and / or mobilization of stem cells from blood or tissue-specific microenvironments, using, for example, MayDay polypeptides as contemplated herein, can utilize autologous or allogeneic mesenchymal stem cells to provide the therapeutic benefits of therapies without the need for donors, cell extraction, ex vivo culture, and transplantation.

[0100] Administration of MayDay polypeptides has been shown to increase stem cell concentrations in lung tissue [Dempsey, SG, CH Miller, J. Schueler, RWF Veale, DJ Day and BCH May (2020). "A novel chemotactic factor derived from the extracellular matrix protein decorin recruits mesenchymal stromal cells in vitro and in vivo." PLoS One 15(7): e0235784]. MayDay polypeptides have also been shown to mobilize endogenous stem cells. Therapeutic administration of MayDay polypeptides can result in therapeutic enhancement of stem cells in the appropriate tissue associated with the disease, thereby negating the need for ex vivo allogeneic or autologous stem cells.

[0101] Again, without wishing to be bound by any theory, Applicant believes that local mobilization of stem cells is broadly beneficial for soft tissue repair and for the treatment, prevention, management, or intervention of respiratory inflammation and / or diseases or conditions associated with respiratory inflammation.

[0102] Without wishing to be bound by any theory, the methods contemplated herein are, in some instances, directed to modulating one or more cellular responses in a subject to which a MayDay polypeptide as contemplated herein is administered.

[0103] In one example, the one or more cellular responses are up-regulation or down-regulation of cytokine expression or modulation of the responsiveness of one or more cells to one or more cytokines.

[0104] In one example, the one or more cellular responses are an upregulation or downregulation of immune cell activation, or a modulation of the responsiveness of one or more immune cells to the upregulation or downregulation.

[0105] In one example, the one or more cellular responses is stem cell activation or mobilization, for example, mesenchymal stem cell activation or mobilization.

[0106] In various examples, a method for treating or preventing respiratory inflammation in a subject in need thereof comprises administering a MayDay polypeptide as contemplated herein in an amount effective to reduce the amount or concentration of one or more pro-inflammatory cytokines, for example, the method comprises administering a MayDay polypeptide in an amount effective to reduce the amount or concentration of a cytokine selected from the group consisting of tumor necrosis factor-α, interferon-γ, and IL-6.

[0107] In various examples, a method for treating or preventing a disease or condition associated with respiratory inflammation in a subject in need thereof comprises administering a MayDay polypeptide as contemplated herein in an amount effective to reduce the amount or concentration of one or more pro-inflammatory cytokines. For example, the method comprises administering a MayDay polypeptide in an amount effective to reduce the amount or concentration of a cytokine selected from the group consisting of tumor necrosis factor-α, interferon-γ, and IL-6.

[0108] In various examples, a method for treating or preventing respiratory inflammation in a subject in need thereof comprises administering a MayDay polypeptide as contemplated herein in an amount effective to reduce the number, activity, or recruitment of immune cells, for example, the recruitment of one or more immune cells to the lungs. For example, the method comprises administering a MayDay polypeptide in an amount effective to reduce the number, activity, or recruitment of immune cells selected from the group consisting of macrophages, neutrophils, beta cells, and dendritic cells, for example, the recruitment of one or more immune cells to the lungs. In a particularly contemplated example, the method comprises administering a MayDay polypeptide in an amount effective to reduce the recruitment of neutrophils to the lungs. In another particularly contemplated example, the method comprises administering a MayDay polypeptide in an amount effective to reduce the number or activity of neutrophils in the lungs.

[0109] In various examples, a method for treating or preventing a disease or condition associated with respiratory inflammation in a subject in need thereof comprises administering a MayDay polypeptide as contemplated herein in an amount effective to reduce the number, activity, or recruitment of immune cells, for example, the recruitment of one or more immune cells to the lungs. For example, the method comprises administering a MayDay polypeptide in an amount effective to reduce the number, activity, or recruitment of immune cells selected from the group consisting of macrophages, neutrophils, beta cells, and dendritic cells, for example, the recruitment of one or more immune cells to the lungs. In a particularly contemplated example, the method comprises administering a MayDay polypeptide in an amount effective to reduce the recruitment of neutrophils to the lungs. In another particularly contemplated example, the method comprises administering a MayDay polypeptide in an amount effective to reduce the number or activity of neutrophils in the lungs.

[0110] In particularly contemplated instances, the decrease is a statistically significant decrease, for example a statistically significant decrease compared to an untreated control.

[0111] Compositions and pharmaceuticals In one aspect, the present invention relates to a pharmaceutical composition comprising an effective amount of a MayDay polypeptide as contemplated herein, e.g., an effective amount of a MayDay polypeptide described herein, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

[0112] Although the following discussion focuses on pharmaceutical compositions, those skilled in the art will understand that the discussion also applies to the preparation of medicaments for the treatment of the diseases or conditions contemplated herein.

[0113] A pharmaceutical composition may contain effective amounts of two or more agents in combination, for example two or more MayDay polypeptides described herein.

[0114] Compositions suitable for the administration of bioactive agents such as bioactive proteins, including therapeutic administration of bioactive polypeptides to subjects in need thereof, are known in the art.

[0115] For example, a biologically active MayDay polypeptide as described herein, or a fragment, variant, peptide analog, or derivative thereof, may in some instances be formulated with a pharmaceutically acceptable carrier, excipient, or combined with other agents to improve the bioavailability, half-life, or efficacy of the biologically active substance, and the composition administered to the subject.

[0116] The term "pharmaceutically acceptable carrier" refers to a carrier (adjuvant or vehicle) that may be administered to a subject with a biologically active substance, such as a MayDay peptide described herein or a pharmaceutically acceptable salt or solvate thereof.

[0117] Traditionally, such a carrier has been saline (0.9% sodium chloride), but other carriers are also suitable.

[0118] Pharmaceutically acceptable carriers that may be used in the compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS), such as d-α-tocopherol polyethylene glycol 1000 succinate, surfactants used in pharmaceutical dosage forms, such as Tweens or other similar polymeric delivery matrices, serum proteins, such as human serum albumin, buffer substances, such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat.

[0119] Cyclodextrins, such as α-, β-, and γ-cyclodextrin, or chemically modified derivatives, such as hydroxyalkyl cyclodextrins (including 2- and 3-hydroxypropyl-3-cyclodextrin), or other solubilizing derivatives, may also be advantageously used to enhance delivery. Oily solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, or carboxymethylcellulose or similar dispersing agent, commonly used in formulating pharmaceutically acceptable dosage forms such as emulsions and / or suspensions.

[0120] The compositions are formulated to allow administration to a subject by any selected route, including, but not limited to, oral or parenteral (including topical, subcutaneous, intramuscular, and intravenous) administration.

[0121] For example, the compositions may be formulated using appropriate pharmaceutically acceptable carriers (including excipients, diluents, adjuvants, and combinations thereof) selected with regard to the intended route of administration and standard pharmaceutical practice.

[0122] Thus, in various examples, the carrier is selected from the group comprising a diluent or excipient.

[0123] In various examples, the compositions may be administered orally as a powder, liquid, tablet, or capsule, or topically as an ointment, cream, or lotion. Suitable formulations may contain additional agents, such as emulsifiers, antioxidants, flavorings, or coloring agents, as needed, and may be adapted for immediate, delayed, modified, sustained, pulsed, or sustained release.

[0124] In certain instances involving administration of a polypeptide as described herein, administration will typically involve direct injection or deposition at the site of interest, e.g., at the site of injury in soft tissue. Parenteral administration is contemplated in some circumstances.

[0125] The compositions may be formulated to optimize bioavailability or activity, or to maintain plasma, blood, or tissue concentrations within a therapeutic range, e.g., for extended periods of time. Sustained release delivery formulations may also be used, e.g., to optimize the concentration of a bioactive agent at the site of action.

[0126] The compositions may be formulated for periodic administration, for example to provide sustained exposure.

[0127] The composition may be administered parenterally. Examples of parenteral dosage forms include the active substance in aqueous solution, isotonic saline, or 5% glucose, or other well-known pharmaceutically acceptable excipients. For example, cyclodextrins or other solubilizing agents well known to those skilled in the art can also be used as pharmaceutical excipients for the delivery of therapeutic agents.

[0128] Examples of dosage forms suitable for oral administration include, but are not limited to, tablets, capsules, lozenges, or similar dosage forms, or any liquid dosage form, such as syrup, aqueous solution, emulsion, etc., that can provide a therapeutically effective amount of the composition. Capsules may contain any standard pharmaceutically acceptable material, such as gelatin or cellulose. Tablets can be prepared by compressing a mixture of active ingredients, solid carriers, and lubricants according to conventional procedures. Examples of solid carriers include starch and sugar bentonite.

[0129] The active ingredient can also be administered in the form of a hard-shell tablet or capsule containing a binder, such as lactose or mannitol, a conventional filler, and a tableting agent. Dosage forms for oral administration can be formulated with an enteric coating to prevent dissolution or disintegration of the dosage form in the stomach, provide delayed release of the drug, and / or allow release of the material after the stomach (e.g., in the upper intestinal tract).

[0130] Examples of dosage forms suitable for transdermal administration include, but are not limited to, transdermal patches, transdermal bandages, and the like.

[0131] Examples of dosage forms suitable for topical administration of the composition include any lotion, stick, spray, ointment, paste, cream, gel, etc., whether applied directly to the skin or via an intermediate such as a pad or patch.

[0132] Examples of dosage forms suitable for administration of the composition in a suppository include any solid dosage form adapted for insertion into a body orifice, particularly those adapted for insertion into the rectum, vagina, and urethra.

[0133] Examples of dosage forms suitable for injection of the composition include delivery via a bolus, such as single or multiple doses by intravenous injection, subcutaneous, intradermal, and intramuscular administration, or oral administration.

[0134] Examples of dosage forms suitable for sustained injectable administration of the composition include pellets or solid dosage forms of the peptide, in which the peptide is encapsulated in a biodegradable polymer matrix, a microemulsion, a liposome, or is encapsulated in a microcapsule.

[0135] Examples of infusion devices for the compositions include infusion pumps that provide desired doses or steady state administration, and include implantable drug pumps.

[0136] Examples of implantable infusion devices for the compositions include any solid dosage form in which a biologically active agent is encapsulated or dispersed within a biodegradable or synthetic polymer, such as silicone, silicone rubber, silastic, or similar polymers.

[0137] Examples of dosage forms suitable for transmucosal delivery of the composition include depository solutions for enemas, pessaries, tampons, creams, gels, pastes, foams, spray solutions, powders, and similar formulations containing, in addition to the active ingredient, carriers known in the art to be appropriate. Such dosage forms include dosage forms suitable for inhalation or instillation of the composition, including compositions comprising solutions and / or suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, and / or powders. Transmucosal administration of the composition may utilize any mucous membrane, but typically utilizes nasal, buccal, vaginal, and rectal tissues. Formulations suitable for intranasal administration of the composition can be administered in liquid form, such as intranasal sprays, nasal drops, or by aerosol administration via a nebulizer, including aqueous or oily solutions of polymer particles. The formulations may be prepared, for example, as aqueous solutions in saline, solutions utilizing benzyl alcohol or other suitable preservatives, absorption enhancers to enhance bioavailability, fluorocarbons and / or other solubilizing or dispersing agents known in the art.

[0138] Examples of dosage forms suitable for buccal or sublingual administration of the composition include lozenges, tablets, etc. Examples of dosage forms suitable for intraocular administration of the composition include solutions and / or suspensions comprising the insert and / or composition in a pharmaceutically acceptable aqueous or organic solvent.

[0139] Examples of formulations of the composition can be found, for example, in Sweetman, SC (ed.), Martindale. The Complete Drug Reference, 33rd Edition, Pharmaceutical Press, Chicago, 2002, 2483 pp.; Aulton, ME (ed.), Pharmaceutics. The Science of Dosage Form Design. Churchill Livingstone, Edinburgh, 2000, 734 pp.; and Ansel, H. C, Allen, LV and Popovich, NG Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Edition, Lippincott 1999, 676 pp. Excipients used in the manufacture of drug delivery systems are described in various publications known to those skilled in the art, including, for example, Kibbe, EH Handbook of Pharmaceutical Excipients, 3rd Edition, American Pharmaceutical Association, Washington, 2000, 665 pp. The United States Pharmacopoeia also provides examples of modified-release oral dosage forms, including those formulated as tablets or capsules. See, for example, United States Pharmacopeia 23rd Edition / National Formulary 18, United States Pharmacopeia, Rockville, MO, 1995 (hereinafter "USP"), which also describes specific tests for determining the drug release potential of extended- and delayed-release tablets and capsules. The USP test for drug release of extended- and delayed-release products is based on drug dissolution from the dosage unit over the elapsed test time. Descriptions of various test equipment and procedures can be found in the US Pharmacopeia.Further guidance on the analysis of extended-release dosage forms is provided by the U.S. Food and Drug Administration (see Guidance for Industry, Extended-release oral dosage forms: development, evaluation, and application of in vitro / in vivo correlations, Rockville, MO; FDA Center for Drug Evaluation and Research, U.S. Food and Drug Administration, 1997).

[0140] When two or more agents are administered or used, the two or more agents may be administered or used simultaneously, sequentially, or separately.

[0141] The invention is further described with reference to the following examples, it being understood that the invention as claimed is not intended to be limited in any way by these examples.

[0142] Example Example 1: Therapeutic efficacy of MayDay polypeptides This example presents an evaluation of the therapeutic efficacy of administration of a MayDay polypeptide as described herein in a mouse model of respiratory inflammation.

[0143] Materials and Methods Test animals All animal experiments were conducted in accordance with guidelines to reduce the amount of suffering, pain, and discomfort in experimental animals. A total of 54 male C57BL / 6 mice were used in this study. Mice, 7–8 weeks old, were obtained from an animal supplier approved by the Institutional Animal Care and Use Committee (IACUC). Animals were acclimatized at least 4 days before the study. Animals were housed in a temperature-controlled room with a 12-hour light / dark cycle and had free access to water and irradiated food throughout the study. Animals were individually identified by ear tag. One day before the start of the study, animals were weighed and stratified into six groups of nine animals each, with each group having approximately the same average weight. Animals were monitored for any form of discomfort, as well as clinical signs of distress or temporary discomfort.

[0144] Study design A total of 54 male C57BL / 6 mice (7-8 weeks old, weighing 220±20 g) were used in this study, as shown in Table 2 below.

[0145] [Table 2]

[0146] Expression and purification of recombinant MayDay polypeptide MayDay, a representative recombinant MayDay polypeptide containing amino acids 31-170 of ovine decorin. 31~170 (MD 31~170 ) was expressed in BL21 E. coli using the pSUMO vector. After amplification, expression, and lysis, the supernatant was loaded onto Q Sepharose™ for purification. Purity (>85%) was confirmed by SDS-PAGE.

[0147] In vivo respiratory inflammation model At -1 hour (1 hour before lipopolysaccharide (LPS) administration), animals were administered vehicle (200 μL) via the intravenous route for groups 1 and 2; dexamethasone (5 mg / kg) (5 mg / kg, 200 μL) as a positive control (group 3) via the intraperitoneal (IP) route; and the appropriate dose (200 μL) of MDV via the intravenous (IV) route for groups 4 to 6. 31~170 was treated with.

[0148] After administration of anesthesia with sodium pentobarbital, mice received a nonlethal intratracheal instillation of 5 mg / kg of lipopolysaccharide O111:B4 (Sigma-Aldrich, St. Louis, MO, USA) derived from Escherichia coli dissolved in 100 μL of phosphate-buffered saline to induce acute respiratory inflammation. Lyophilized powder of lipopolysaccharide (Sigma L4391, O111:B4) was resuspended in 0.9% saline to obtain 2.5 mg / mL aliquots, which were stored at 20°C. At time 0, animals in vehicle group 1 were administered 40 μL of phosphate-buffered saline via the intratracheal route, and animals in groups 2–6 were administered 40 μL of lipopolysaccharide at 5 mg / kg via the intratracheal route to induce acute respiratory inflammation. The study was terminated at either 6, 24 or 72 hours, at which point each animal was weighed.

[0149] Lung harvest and bronchoalveolar lavage fluid collection Lungs were collected from each animal and weighed. Lungs were flushed twice with 0.5 mL of Hank's balanced salt solution; both 0.5 mL lavage fluids were immediately pooled and kept on ice until processing. Bronchoalveolar lavage fluid was centrifuged to pellet cells, divided into four aliquots, and stored at -80°C until Mesoscale Discovery (MSD) analysis. The cell pellet was resuspended in 100 μL of phosphate-buffered saline, and total cell counts were determined using a Cell Counter model R1 (Olympus, Tokyo, Japan). Cells were then stained with Wright-Giemsa stain (Sora Bio, Beijing, China) according to the manufacturer's protocol. Neutrophil counts were quantified using a light microscope by counting a total of 200 cells per slide at 40x magnification.

[0150] Whole lungs were fixed in 10% neutral buffered formalin (NBF), embedded in paraffin, and then cut into 4 μm sections, which were then deparaffinized and stained with hematoxylin and eosin (H&E) for histological evaluation.

[0151] Measurement of inflammatory cytokines Bronchoalveolar lavage fluid cytokine concentrations were assessed from bronchoalveolar lavage fluid aliquots using a 29-plex cytokine MSD platform (product number K152167D) according to the manufacturer's instructions.

[0152] Myeloperoxidase levels in bronchoalveolar lavage fluid Myeloperoxidase levels from aliquots of bronchoalveolar lavage fluid were assessed by myeloperoxidase assay using an Abcam kit (product no. Ab155458) according to the manufacturer's instructions.

[0153] Assessment of lung histology The severity of lung tissue inflammation was assessed using the lung injury score as follows: The severity of lung injury was scored based on the following histological features: alveolar congestion, hemorrhage, neutrophil infiltration into airspaces or vascular walls, and alveolar wall thickness / hyaline membrane formation. Each item was graded on a 5-point scale from 0 to 4: 0 (minimal injury), 1 (mild injury), 2 (moderate injury), 3 (severe injury), and 4 (maximal injury).

[0154] statistical analysis Figures were prepared and statistical analysis was performed using GraphPad Prism software (San Diego, USA). All data were presented as mean ± standard deviation (SD). Statistical comparisons between pairs of groups were determined by two-tailed Student's t-test. The P value for significance was set at 0.05.

[0155] result MayDay polypeptide reduces lung inflammation To explore the therapeutic potential of representative MayDay polypeptides in ameliorating respiratory inflammation in vivo, a mouse model was established by intratracheal administration of lipopolysaccharide. Hematoxylin and eosin staining was performed for histological examination of the lungs.

[0156] After induction of respiratory inflammation with lipopolysaccharide (5 mg / kg) (Group 2), acute inflammatory responses, such as intact alveolar walls, interstitial edema, and infiltration of inflammatory cells, were observed in lung tissue (data not shown).

[0157] In particular, the above pathological changes in the lungs are 31~170 was alleviated in subjects receiving MD (Groups 4, 5, and 6). 31~170 The positive effects of administration of acetaminophen were comparable to or better than those observed with the positive control, dexamethasone (Group 3). See, e.g., Figures 7-9.

[0158] Cytokine analysis MSD analysis is 31~170 As shown in Figure 2A, it significantly reduced the protein levels of tumor necrosis factor-α in bronchoalveolar lavage fluid 6 hours after administration compared with the lipopolysaccharide control group (Group 2), and as shown in Figures 3B and 4B, it significantly reduced the protein levels of IFN-γ and IL-6 in bronchoalveolar lavage fluid 24 hours after administration compared with the lipopolysaccharide group (Group 2), respectively.

[0159] These data are 31~170 administration caused a rapid and significant decrease in the pro-inflammatory cytokine TNF-α and a (slightly slower) significant decrease in the pro-inflammatory cytokines IFN-γ and IL-6 after challenge with lipopolysaccharide in this mouse model of respiratory inflammation.

[0160] Cell counts and differentials in bronchoalveolar lavage fluid Furthermore, as can be clearly seen in Figure 5, the number of neutrophils also increased with increasing MD in mice. 31~170 Furthermore, myeloperoxidase (MPO), an indicator of neutrophils and inflammation, was significantly reduced at all evaluated time points after MD administration. 31~170 The levels of α-glucan in the α-glucan-treated animals (groups 4, 5 and 6) were significantly reduced at 6 and 24 hours (Figure 6).

[0161] These data are 31~170 We show that administration of acetaminophen caused a rapid and significant decrease in cellular inflammatory mediators, namely, neutrophil numbers and myeloperoxidase levels, after lipopolysaccharide challenge in this mouse model of respiratory inflammation. Myeloperoxidase is produced by activated neutrophils as part of the inflammatory response and is established as an early indicator of inflammation.

[0162] Tissue diagnosis Lung injury score is MD 31~170 In mice treated with MD 31~170 The injury in animals treated with MD (Groups 4, 5, and 6) was comparable to or less than that observed in animals treated with dexamethasone (Group 3) (see Figures 7-9, MD (2.5 μg, 25 μg, and 100 μg) vs. dexamethasone). Indeed, statistically significant reductions in neutrophil scores, inflammation scores, and interstitial inflammation scores were observed in Group 6 (MD) compared to the lipopolysaccharide control (Group 2), as shown in Figures 7B, 8B, and 9B, respectively. 31~170 , 100 μg / mouse) at 24 hours.

[0163] Histological analysis of this lung tissue was performed by MD 31~170 showed that administration of α-glucan 1-hydroxybenzoate induced dose-dependent changes in neutrophil, inflammation, and interstitial inflammation scores 6, 24, and 72 hours after lipopolysaccharide challenge in this mouse model of respiratory inflammation.

[0164] Consideration These results demonstrate that representative MayDay polypeptides can effectively attenuate acute respiratory inflammation in animal models, which in turn supports the use of MayDay polypeptides in methods of treating or preventing respiratory inflammation, and diseases or conditions associated with respiratory inflammation.

[0165] Publications [Table 3]

[0166] The words "comprise", "comprises", "including" and similar words used herein should not be construed in an exclusive or exhaustive sense. In other words, they are intended to mean "including but not limited to". When interpreting each statement herein that includes the term "comprise", "comprises", or "including", there may be features other than those prefaced by the term.

[0167] The entire disclosures of all applications, patents and publications, cited above and below (if any), are hereby incorporated by reference.

[0168] Where reference is made in the foregoing description to integers or components that have known equivalents thereof, such integers are incorporated herein as if individually set forth.

[0169] It should be noted that various changes and modifications to the presently preferred examples described herein will be apparent to those skilled in the art. Such changes and modifications can be made without departing from the spirit and scope of the present invention and without diminishing its attendant advantages. Therefore, such changes and modifications are intended to be included in the present invention.

[0170] The invention may also be broadly stated to consist of any and all combinations of two or more of the parts, elements and features, individually or collectively, referenced or shown in the application specification.

[0171] Aspects of the invention have been described by way of example only, and it should be understood that changes, modifications, and additions may be made thereto without departing from the scope of the invention, e.g., as defined in claims (indicative or otherwise), where such claims exist. Furthermore, where known equivalents exist for particular features, such equivalents are incorporated as if specifically referred to herein.

[0172] Sequence Listing [ka] TIFF2026504985000007.tif245169 TIFF2026504985000008.tif245169 TIFF2026504985000009.tif241169 TIFF2026504985000010.tif59169

Claims

1. 20. A method of treating or preventing respiratory inflammation in a subject in need thereof, comprising administering to said subject a therapeutically effective amount of a MayDay polypeptide.

2. 10. The method of claim 1, wherein the subject is suffering from, or is at increased risk of suffering from, or is predicted to be at increased risk of suffering from, a disease or condition associated with respiratory inflammation.

3. 10. A method of treating or preventing a disease or condition associated with respiratory inflammation in a subject in need thereof, comprising administering to said subject a therapeutically effective amount of a MayDay polypeptide.

4. Diseases or conditions associated with respiratory inflammation include, but are not limited to, acute respiratory distress syndrome (ARDS) (including pathogen-induced ARDS, smoke-induced ARDS, pneumonia-induced ARDS, sepsis-induced ARDS, inhalation injury-induced ARDS (including ARDS associated with inhalation of water, vomit, smoke, or chemicals), immune response-induced ARDS (such as transfusion-related ARDS), and acute pancreatitis-induced ARDS); sepsis; chronic obstructive pulmonary disease; interstitial lung disease; asthma (including, but not limited to, eosinophilic asthma, neutrophilic asthma, granulocyte-poor asthma, and mixed granulocyte asthma). ), pulmonary fibrosis; asbestosis; interstitial pneumonia; pneumonitis (including but not limited to lymphocytic interstitial pneumonia and hypersensitivity pneumonitis); collagen vascular disease; granulomatous vasculitis; sarcoidosis; cystic fibrosis; drug-induced pulmonary disease (including but not limited to toxin-induced pulmonary disease (such as pulmonary disease associated with methotrexate, bleomycin, etc.) and immune-mediated pulmonary disease (such as pulmonary disease associated with pembrolizumab)); bronchiectasis; bronchitis; bronchiolitis; and bronchiolitis obliterans.

5. 5. The method of claim 4, wherein the disease or condition is selected from the group consisting of acute respiratory distress syndrome, chronic obstructive pulmonary disease, and asthma.

6. A method for treating or preventing acute respiratory distress syndrome and / or chronic obstructive pulmonary disease and / or asthma, comprising administering to a subject a therapeutically effective amount of a MayDay polypeptide.

7. The MayDay polypeptide is a) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 1-188 of mammalian decorin; b) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31-188 of mammalian decorin; c) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 1-188 of mammalian decorin; d) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 consecutive amino acids corresponding to any amino acid sequence within residues 31-188 of mammalian decorin; e) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 1-177 of mammalian decorin; f) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31-177 of mammalian decorin; g) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 1-177 of mammalian decorin; h) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 contiguous amino acids corresponding to any amino acid sequence within residues 31-177 of mammalian decorin; i) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 1-170 of mammalian decorin; j) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31-170 of mammalian decorin; k) a polypeptide comprising, consisting essentially of, or consisting of an amino acid sequence corresponding to residues 31-169 of human decorin; l) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 consecutive amino acids corresponding to any amino acid sequence within residues 1-170 of mammalian decorin; m) an N-terminal fragment of mammalian decorin comprising, consisting essentially of, or consisting of at least 10 consecutive amino acids corresponding to any amino acid sequence within residues 31-170 of mammalian decorin; n) a polypeptide comprising, consisting essentially of, or consisting of the amino acid sequence set forth in any one of SEQ ID NOs: 1 to 13; o) a polypeptide comprising, consisting essentially of, or consisting of at least about 10 contiguous amino acids from any one of a) through n) above; p) a polypeptide comprising or consisting of at least about 10 consecutive amino acids from any one of a) to o) above, wherein the polypeptide comprises a motif or region capable of interacting with or recruiting one or more cells, including one or more stem cells; q) a polypeptide comprising or consisting of at least about 10 consecutive amino acids from any one of a) to p) above, wherein said polypeptide comprises a motif or region capable of interacting with or recruiting one or more mesenchymal stem cells; r) a functional fragment, functional variant, peptide analog, or peptidomimetic, or derivative of any one of a) to q) above; s) a polypeptide having at least about 90% amino acid identity to any one of a) to r) above; t) Any combination of two or more of the above a) to s) 7. The method of any one of claims 1 to 6, wherein the polypeptide is an isolated, purified, recombinant, or synthetic polypeptide selected from the group comprising:

8. A method for mediating a biological effect in a subject suffering from or susceptible to respiratory inflammation, the method comprising administering to the subject an effective amount of a polypeptide as defined in claim 7.

9. A method for modulating tissue repair in a subject suffering from or susceptible to respiratory inflammation, the method comprising administering to the subject a therapeutically effective amount of a polypeptide as defined in claim 7.

10. A method for modulating stem cell mobilization or related processes in a subject suffering from or susceptible to respiratory inflammation, the method comprising administering to the subject a therapeutically effective amount of a polypeptide as defined in claim 7.

11. 11. The method of claim 10, wherein the associated process is selected from the group consisting of angiogenesis, hematopoiesis, protein expression, induction or deposition, tissue remodeling, repair or regeneration, cell proliferation, cell differentiation (including stem cell differentiation), cell regulation, apoptosis, modulation of one or more immune responses, tumorigenesis, chemotaxis, or modulation of cell recruitment.

12. 12. The method of any one of claims 8 to 11, wherein the subject has, or is at increased risk of, or is predicted to be at increased risk of, a disease or condition associated with respiratory inflammation.

13. A pharmaceutical composition for treating or preventing respiratory inflammation and / or diseases or conditions associated with respiratory inflammation, the pharmaceutical composition comprising one or more of the polypeptides defined in claim 7.

14. A pharmaceutical composition for treating or preventing acute respiratory distress syndrome and / or chronic obstructive pulmonary disease and / or asthma, said pharmaceutical composition comprising one or more of the polypeptides defined in claim 7.

15. 10. A polypeptide as defined in claim 7 for treating respiratory inflammation and / or a disease or condition associated with respiratory inflammation in a subject in need thereof.

16. 10. A polypeptide as defined in claim 7 for treating acute respiratory distress syndrome and / or chronic obstructive pulmonary disease and / or asthma in a subject in need thereof.

17. 8. A polypeptide as defined in claim 7 for mediating a biological effect in a subject suffering from or susceptible to respiratory inflammation and / or a disease or condition associated with respiratory inflammation, for modulating stem cell mobilization or related processes in the subject, or for modulating tissue repair in the subject.

18. 10. Use of a polypeptide as defined in claim 7 in the preparation of a medicament for the treatment of respiratory inflammation and / or diseases or conditions associated with respiratory inflammation.

19. 10. Use of a polypeptide as defined in claim 7 in the preparation of a medicament for the treatment of acute respiratory distress syndrome and / or chronic obstructive pulmonary disease and / or asthma.

20. 10. Use of a polypeptide as defined in claim 7 in the preparation of a medicament for mediating a biological effect in a subject suffering from or susceptible to respiratory inflammation and / or a disease or condition associated with respiratory inflammation, for modulating stem cell mobilization or related processes in the subject, or for modulating tissue repair in the subject.

21. 21. The method, composition, polypeptide or use of any preceding claim, wherein said polypeptide comprises, consists essentially of, or consists of an amino acid sequence corresponding to the amino acid sequence set forth in SEQ ID NO: 6 or 7.

22. 22. The method, composition, polypeptide or use of any preceding claim, wherein said polypeptide comprises, consists essentially of, or consists of an amino acid sequence corresponding to the amino acid sequence set forth in SEQ ID NO:

2.

23. 23. The method, composition, polypeptide or use of any of claims 1 to 22, wherein said polypeptide comprises, consists essentially of, or consists of an amino acid sequence corresponding to the amino acid sequence set forth in any one of SEQ ID NOs: 3, 4, 9, 10, 12 or 13.

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