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 whey acidic protein (WAP) domain containing polypeptide or a sequence derived from a WAP domain 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 for inhibiting serine proteases having at least one human alpha-1 antitrypsin (AAT) polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 32, and SEQ ID NO: 33, wherein the AAT polypeptide is operably linked to an immunoglobulin Fc polypeptide having an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 6.
2. The isolated fusion protein of claim 1 , 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.
3. The isolated fusion protein of claim 2 , wherein the hinge region or linker region comprises the amino acid sequence of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, or SEQ ID NO: 46.
4. The isolated fusion protein of claim 1 , wherein the human AAT polypeptide comprises 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.
5. The isolated fusion protein of claim 1 , wherein the human AAT polypeptide comprises the amino acid sequence of SEQ ID NO: 2 operably linked to an immunoglobulin Fc polypeptide comprising the amino acid sequence of SEQ ID NO: 6.
6. The isolated fusion protein of claim 1 , wherein the human AAT polypeptide comprises the amino acid sequence of SEQ ID NO: 32 operably linked to an immunoglobulin Fc polypeptide comprising the amino acid sequence of SEQ ID NO: 6.
7. The isolated fusion protein of claim 1 , wherein the human AAT polypeptide comprises the amino acid sequence of SEQ ID NO: 33 operably linked to an immunoglobulin Fc polypeptide comprising the amino acid sequence of SEQ ID NO: 6.
8. The isolated fusion protein of claim 1 , wherein the fusion protein further comprises a whey acidic protein (WAP) domain-containing polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 9-13.
9. The isolated fusion protein of claim 4 , wherein the fusion protein further comprises a secretory leukocyte protease inhibitor (SLPI) polypeptide comprising the amino acid sequence of SEQ ID NO: 9 or SEQ ID NO: 10.
10. The isolated fusion protein of claim 1 , wherein the immunoglobulin Fc polypeptide is modified to enhance FcRn binding.
11. 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:
a Met to Tyr mutation corresponding to amino acid residue at position 22 of SEQ ID NO: 6;
a Ser to Thr mutation corresponding to amino acid residue at position 24 in SEQ ID NOs: 6;
a Thr to Glu mutation corresponding to amino acid residue at position 26 in SEQ ID NO: 6;
a Met to Leu mutation corresponding to amino acid residue at position 198 in SEQ ID NO: 6; and
a Asn to Ser mutation corresponding to amino acid residue at position 204 in SEQ ID NO: 6.
12. A method of inhibiting or downregulating aberrant serine protease expression or activity in a subject in need thereof, the method comprising administering a fusion protein according to claim 1 .
13. The method of claim 12 , wherein the subject is a human.
14. The method of claim 13 , wherein the subject has a disease or disorder 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.
15. The method of claim 13 , wherein the subject has an infection selected from the following: bacterial infections, fungal infections, or viral infections.