Therapeutic nuclease compositions and methods
Hybrid nuclease molecules and methods for treating an immune-related disease or disorder in a mammal, and a pharmaceutical composition for treating an immune-related disease in a mammal.
1. A polypeptide consisting of human RNase 1 operatively coupled with or without a linker to a mutant human IgG1 Fc domain, wherein the Fc domain comprises a P238S mutation and a P331S mutation, numbering according to the EU index, and wherein the Fc domain does not contain a variable region that binds antigen.
2. The polypeptide of claim 1 , wherein the Fc domain has decreased binding to Fcγ receptors on human cells.
3. The polypeptide of claim 1 which has at least 1, 2, 3, 4, or 5-fold less cytotoxicity relative to a polypeptide having wild type Fc domain.
4. The polypeptide of claim 1 which has an increased serum half-life relative to a polypeptide comprising human RNase 1 without an Fc domain.
5. The polypeptide of claim 1 which degrades circulating RNA and RNA in immune complexes, or inhibits interferon-γ production, or both.
6. The polypeptide of claim 1 , wherein the Fc domain comprises a hinge domain, a CH2 domain and a CH3 domain.
7. The polypeptide of claim 1 , wherein the Fc domain further comprises a substitution of one or more of three hinge region cysteine residues with serine.
8. The polypeptide of claim 1 , wherein the Fc domain further comprises an additional mutation selected from the group consisting of SCC, SSS (residues 220, 226, and 229), G236R, L328R, L234A, and L235A, numbering according to the EU index.
9. The polypeptide of claim 1 , wherein the Fc domain further comprises an SCC mutation (residues 220, 226, and 229), numbering according to the EU index.
10. The polypeptide of claim 1 , wherein the human RNase 1 is linked to the N-terminus of the Fc domain.
11. The polypeptide of claim 1 , wherein the human RNase 1 is linked to the C-terminus of the Fc domain.
12. The polypeptide of claim 1 , wherein the human RNase 1 is operatively coupled to the Fc domain without a linker domain.
13. The polypeptide of claim 1 , wherein the human RNase 1 is operatively coupled to the Fc domain via a linker domain.
14. The polypeptide of claim 13 , wherein the linker domain is a polypeptide linker.
15. The polypeptide of claim 14 , wherein the polypeptide linker is a gly-ser linker.
16. The polypeptide of claim 1 , wherein the human RNase 1 comprises the amino acid sequence set forth in SEQ ID NO: 101.
17. The polypeptide of claim 1 , wherein the Fc domain comprises the amino acid sequence set forth in SEQ ID NO: 90.
18. A polypeptide consisting of human RNase 1 operatively coupled without a linker to the N-terminus of a mutant human IgG1 Fc domain, wherein the Fc domain comprises an SCC mutation (residues 220, 226, and 229), a P238S mutation, and a P331S mutation, numbering according to the EU index, and wherein the Fc domain does not contain a variable region that binds antigen.
19. The polypeptide of claim 18 , wherein the Fc domain comprises a hinge domain, a CH2 domain and a CH3 domain.
20. The polypeptide of claim 18 , wherein the human RNase 1 comprises the amino acid sequence set forth in SEQ ID NO: 101.
21. The polypeptide of claim 18 , wherein the Fc domain comprises the amino acid sequence set forth in SEQ ID NO: 90.
22. A polypeptide consisting of the amino acid sequence set forth in SEQ ID NO: 96.
23. A dimer comprising a polypeptide of claim 1 .
24. A dimer comprising a polypeptide of claim 16 .
25. A dimer comprising a polypeptide of claim 18 .
26. A dimer comprising a polypeptide of claim 22 .
27. A composition comprising the polypeptide of claim 1 and a pharmaceutically acceptable carrier.
28. A composition comprising a dimer of claim 23 and a pharmaceutically acceptable carrier.
29. A polypeptide consisting of human RNase 1 operatively coupled without a linker to the C-terminus of a mutant human IgG1 Fc domain, wherein the Fc domain comprises an SCC mutation (residues 220, 226, and 229), a P238S mutation, and a P331S mutation, numbering according to the EU index, and wherein the Fc domain does not contain a variable region that binds antigen.
30. The polypeptide of claim 29 , wherein the Fc domain comprises a hinge domain, a CH2 domain and a CH3 domain.
31. The polypeptide of claim 29 , wherein the human RNase 1 comprises the amino acid sequence set forth in SEQ ID NO: 101.
32. The polypeptide of claim 29 , wherein the Fc domain comprises the amino acid sequence set forth in SEQ ID NO: 90.
33. A dimer comprising the polypeptide of claim 29 .
34. A composition comprising the polypeptide of claim 18 and a pharmaceutically acceptable carrier.
35. A composition comprising a dimer of claim 25 and a pharmaceutically acceptable carrier.
36. A composition comprising the polypeptide of claim 22 and a pharmaceutically acceptable carrier.
37. A composition comprising a dimer of claim 26 and a pharmaceutically acceptable carrier.
38. A composition comprising the polypeptide of claim 29 and a pharmaceutically acceptable carrier.
39. A composition comprising a dimer of claim 33 and a pharmaceutically acceptable carrier.