Serpin fusion polypeptides and methods of use thereof

Abstract

This invention relates to molecules, particularly polypeptides, more particularly fusion proteins that include a serpin polypeptide or an amino acid sequence that is derived from a serpin and second polypeptide comprising of at least one the following: an Fc polypeptide or an amino acid sequence that is derived from an Fc polypeptide; a cytokine targeting polypeptide or a sequence derived from a cytokine targeting polypeptide; a WAP domain containing polypeptide or a sequence derived from a WAP containing polypeptide; and an albumin polypeptide or an amino acid sequence that is derived from a serum albumin polypeptide. This invention also relates to methods of using such molecules in a variety of therapeutic and diagnostic indications, as well as methods of producing such molecules.

What is claimed is: 
     
         1 . An isolated fusion protein comprising at least one human serpin polypeptide operably linked to a human immunoglobulin Fc polypeptide or an amino acid sequence that is derived from an immunoglobulin Fc polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, and 65. 
     
     
         2 . The isolated fusion protein of  claim 1 , wherein the fusion protein further comprises an additional polypeptide selected from the group consisting of:
 a cytokine targeting polypeptide or a sequence derived from a cytokine targeting polypeptide;   a WAP domain containing polypeptide or a sequence derived from a WAP domain containing polypeptide; or   an albumin polypeptide or an amino acid sequence that is derived from a serum albumin polypeptide.   
     
     
         3 . The fusion protein of  claim 1 , wherein the human serpin polypeptide is a human alpha-1 antitrypsin (AAT) polypeptide or is derived from a human AAT polypeptide. 
     
     
         4 . The isolated fusion protein of  claim 3 , wherein AAT polypeptide comprises the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 80. 
     
     
         5 . The isolated fusion protein of  claim 3 , wherein the AAT polypeptide comprises the reactive site loop of AAT comprising the amino acid sequence of SEQ ID NO: 1. 
     
     
         6 . The isolated fusion protein of  claim 3 , wherein the AAT polypeptide comprises a mutated reactive site loop of AAT comprising the amino acid sequence of SEQ ID NO: 32 or 33. 
     
     
         7 . The isolated fusion protein of  claim 1 , wherein the immunoglobulin Fc polypeptide comprises one or more mutations at a position selected from M252, T246, M428, and combinations thereof. 
     
     
         8 . The isolated fusion protein of  claim 1 , wherein the immunoglobulin Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 53. 
     
     
         9 . The isolated fusion protein of  claim 1 , wherein the immunoglobulin Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 73. 
     
     
         10 . The isolated fusion protein of  claim 1 , wherein the serpin polypeptide and the immunoglobulin Fc polypeptide are operably linked via a hinge region, a linker region, or both a hinge region and linker region. 
     
     
         11 . The isolated fusion protein of  claim 3 , wherein the AAT polypeptide and the immunoglobulin Fc polypeptide are operably linked via a hinge region, a linker region, or both a hinge region and linker region. 
     
     
         12 . The isolated fusion protein of  claim 10 , wherein the hinge region, the linker region or both the hinge region and the linker region comprise a peptide sequence. 
     
     
         13 . The isolated fusion protein of  claim 10 , wherein the hinge region, the linker region or both the hinge region and the linker region comprise a peptide sequence. 
     
     
         14 . The isolated fusion protein of  claim 1 , wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 78 or 79. 
     
     
         15 . A method of treating or alleviating a symptom of a disease or disorder associated with aberrant serine protease expression or activity in a subject in need thereof, the method comprising administering a fusion protein according to  claim 1 . 
     
     
         16 . A method of treating or alleviating inflammation or a symptom of an inflammatory disease or disorder while reducing the risk of infection, in a subject in need thereof, the method comprising administering a fusion protein according to  claim 1 . 
     
     
         17 . A method of reducing the risk of infection in a subject in need thereof, the method comprising administering a fusion protein according to  claim 1 . 
     
     
         18 . The method of  claim 17 , wherein the subject is a human. 
     
     
         19 . The method of  claim 17 , wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 78 or 79. 
     
     
         20 . The method of  claim 16 , wherein the inflammatory disease or disorder is selected from the following: emphysema, chronic obstructive pulmonary disease (COPD), acute respiratory distress syndrome (ARDS), allergic asthma, cystic fibrosis, cancers of the lung, ischemia-reperfusion injury, ischemia/reperfusion injury following cardiac transplantation, myocardial infarction, rheumatoid arthritis, septic arthritis, psoriatic arthritis, ankylosing spondylitis, Crohn's disease, psoriasis, type I and/or type II diabetes, pneumonia, sepsis, graft versus host disease (GVHD), wound healing, Systemic lupus erythematosus, and Multiple sclerosis. 
     
     
         21 . The method of  claim 16 , wherein the infection is selected from bacterial infections, fungal infections, or viral infections.