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 alpha-1 antitrypsin (AAT) polypeptide, an Elafin polypeptide, and an immunoglobulin Fc polypeptide, such that at least two of the AAT polypeptide, the Elafin polypeptide, and the immunoglobulin Fc polypeptide are operably linked via a hinge region, a linker region, or both a hinge region and linker region.
2. The fusion protein of claim 1 , wherein the AAT polypeptide is a human alpha-1 antitrypsin (AAT) polypeptide.
3. The isolated fusion protein of claim 2 , wherein AAT polypeptide comprises the amino acid sequence of SEQ ID NO: 2.
4. The isolated fusion protein of claim 2 , wherein the AAT polypeptide comprises the reactive site loop of AAT comprising the amino acid sequence of SEQ ID NO: 1 or a mutated reactive site loop of AAT comprising the amino acid sequence of SEQ ID NO: 32 or 33.
5. The isolated fusion protein of claim 2 , wherein the immunoglobulin Fc polypeptide is a human Fc polypeptide.
6. The isolated fusion protein of claim 5 , wherein human Fc polypeptide is a human IgM polypeptide or a human IgG Fc polypeptide.
7. The isolated fusion protein of claim 6 , wherein the human IgG Fc polypeptide is a human IgG1 polypeptide, a human IgG2 Fc polypeptide, human IgG3 Fc polypeptide, or human IgG4 Fc polypeptide.
8. The isolated fusion protein of claim 1 , wherein the immunoglobulin Fc polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 3, 4, 5, 6, and 7.
9. The isolated fusion protein of claim 1 , wherein the hinge region, the linker region or both the hinge region and the linker region comprise a peptide sequence.
10. The isolated fusion protein of claim 1 , where the fusion protein comprises the amino acid sequence of SEQ ID NO: 29.
11. The isolated fusion protein of claim 1 , wherein the immunoglobulin Fc polypeptide is modified to enhance FcRn binding.
12. The isolated fusion protein of claim 8 , wherein the immunoglobulin Fc polypeptide comprises at least one of the following mutations:
a Met to Tyr mutation corresponding to amino acid residue at position 22 of SEQ ID NOs: 3, 5 and 6, and amino acid residue at position 21 of SEQ ID NO: 4;
a Ser to Thr mutation corresponding to amino acid residue at position 24 in SEQ ID NOs: 3, 5 and 6, and amino acid residue at position 23 in SEQ ID NO: 4;
a Thr to Glu mutation corresponding to amino acid residue at position 26 in SEQ ID NOs: 3, 5 and 6, and amino acid residue at position 25 in SEQ ID NO: 4;
a Met to Leu mutation corresponding to amino acid residue at position 198 in SEQ ID NOs: 3, 5 and 6, and amino acid residue at position 197 in SEQ ID NO: 4; and
a Asn to Ser mutation corresponding to amino acid residue at position 204 in SEQ ID NOs: 3, 5 and 6, and amino acid residue at position 203 in SEQ ID NO: 4.
13. The isolated fusion protein of claim 1 , wherein the Elafin polypeptide is a human Elafin polypeptide.
14. The isolated fusion protein of claim 13 , wherein the human Elafin polypeptide is a full-length Elafin polypeptide.
15. The isolated fusion protein of claim 1 , wherein the Elafin polypeptide comprises an amino acid sequence selected the group consisting of SEQ ID NO: 11, 12, and 13.