Serpin fusion polypeptides and methods of use thereof
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.
1. An isolated fusion protein comprising at least one human serpin polypeptide comprising a human alpha-1 antitrypsin (AAT) polypeptide comprising the amino acid sequence of SEQ ID NO: 1 operably linked to
an immunoglobulin Fc polypeptide comprising the amino acid sequence of SEQ ID NO: 6.
2. The isolated fusion protein of claim 1 , wherein AAT polypeptide comprises the amino acid sequence of SEQ ID NO: 2.
3. 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.
4. The isolated fusion protein of claim 3 , wherein the hinge region, the linker region or both the hinge region and the linker region comprise a peptide sequence.
5. The isolated fusion protein of claim 4 , wherein the peptide sequence comprises the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, or SEQ ID NO: 46.
6. The isolated fusion protein of claim 1 , wherein the fusion protein further comprises a WAP domain-containing polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 9-13.
7. The isolated fusion protein of claim 1 , wherein the immunoglobulin Fc polypeptide is modified to enhance FcRn binding.
8. The isolated fusion protein of claim 1 , wherein the immunoglobulin Fc polypeptide comprises at least one of the following mutations: Met252Tyr, Ser254Thr, Thr256Glu, Met428Leu or Asn434Ser.
9. The isolated fusion protein of claim 1 , wherein the immunoglobulin Fc polypeptide comprises at least one mutation at a position selected from the group consisting of: Met252, Ser254, Thr256, Met428, and Asn434.
10. An isolated fusion protein comprising at least one human alpha-1 antitrypsin (AAT) polypeptide comprising a human alpha-1 antitrypsin (AAT) polypeptide comprising the amino acid sequence of SEQ ID NO: 32 or SEQ ID NO: 33 operably linked to
an immunoglobulin Fc polypeptide comprising the amino acid sequence of SEQ ID NO: 6.
11. The isolated fusion protein of claim 10 , 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 fusion protein further comprises a SLPI polypeptide comprising the amino acid sequence of SEQ ID NO: 9 or SEQ ID NO: 10.
13. The isolated fusion protein of claim 11 , 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 10 , wherein the immunoglobulin Fc polypeptide is modified to enhance FcRn binding.
15. The isolated fusion protein of claim 10 , wherein the immunoglobulin Fc polypeptide comprises at least one mutation at a position selected from the group consisting of: Met252, Ser254, Thr256, Met428, and Asn434.
16. The isolated fusion protein of claim 10 , wherein the immunoglobulin Fc polypeptide comprises at least one of the following mutations: Met252Tyr, Ser254Thr, Thr256Glu, Met428Leu or Asn434Ser.
17. 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 or claim 10 .
18. The method of claim 17 , wherein the subject is a human.
19. 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 or claim 10 .
20. The method of claim 19 , 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. 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 or claim 10 .
22. The method of claim 21 , wherein the infection is selected from bacterial infections, fungal infections, or viral infections.