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 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. A method of inhibiting or downregulating aberrant serine protease expression or activity in a subject in need thereof, the method comprising administering an isolated fusion protein comprising
at least one alpha-1 antitrypsin (AAT) polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 2, 34 or 35,
an Elafin polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 11, 12, and 13, 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 method of claim 1 , wherein the subject is a human.
3. The method of claim 1 , wherein the subject has an inflammatory 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, 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.
4. The method of claim 1 , wherein the subject has an infection selected from bacterial infections, fungal infections, or viral infections.
5. The method of claim 1 , wherein the immunoglobulin Fc polypeptide is a human Fc polypeptide.
6. The method of claim 5 , wherein the human Fc polypeptide is a human IgG1 Fc polypeptide, a human IgG2 Fc polypeptide, human IgG3 Fc polypeptide, or human IgG4 Fc polypeptide.
7. The method of claim 6 , wherein the human Fc polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 3, 4, 5, and 6.
8. The method of claim 1 , wherein the hinge region, the linker region or both the hinge region and the linker region comprise a peptide sequence.
9. The method of claim 1 , where the fusion protein comprises the amino acid sequence of SEQ ID NO: 29.
10. The method of claim 7 , wherein the immunoglobulin Fc polypeptide is modified to enhance FcRn binding.
11. The method of claim 10 , wherein the modified immunoglobulin Fc polypeptide comprises at least one of the following mutations:
a Met to Tyr mutation corresponding to the 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 the 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 the 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 the 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
an Asn to Ser mutation corresponding to the 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.