Interleukin-10 fusion proteins
The present invention generally relates to fusion proteins of antibodies and interleukin-10 (IL-10). In addition, the present invention relates to polynucleotides encoding such fusion proteins, and vectors and host cells comprising such polynucleotides. The invention further relates to methods for producing the fusion proteins of the invention, and to methods of using them in the treatment of disease.
1. A fusion protein comprising an antibody and an IL-10 molecule, wherein the antibody comprises the heavy chain variable region of SEQ ID NO: 67 and the light chain variable region of SEQ ID NO: 69, and wherein the IL-10 molecule comprises SEQ ID NO:1.
2. The fusion protein of claim 1 , wherein the fusion protein comprises two identical heavy chain polypeptides and two identical light chain polypeptides.
3. The fusion protein of claim 2 , wherein each of the heavy chain polypeptides comprises an IgG-class antibody heavy chain and a monomeric IL-10 molecule.
4. The fusion protein of claim 3 , wherein the monomeric IL-10 molecule is fused at its N-terminus to the C-terminus of the IgG-class antibody heavy chain.
5. The fusion protein of claim 4 additionally comprising a peptide linker to fuse the N-terminus of the monomeric IL-10 molecule to the C-terminus of the IgG-class antibody heavy chain.
6. The fusion protein of claim 3 , wherein each of the heavy chain polypeptides further comprises a peptide linker.
7. The fusion protein of claim 3 , wherein the monomeric IL-10 molecules comprised in the heavy chain polypeptides form a functional homodimeric IL-10 molecule.
8. The fusion protein of claim 1 , wherein the antibody is an IgG-class antibody and comprises a modification, which modification reduces binding affinity of the antibody to an Fc receptor, as compared to a corresponding IgG-class antibody without the modification, wherein the IgG-class antibody comprises an amino acid substitution at a position according to EU numbering selected from the group consisting of 228 , 233 , 234 , 235 , 297 , 329 , and 331 of an antibody heavy chain.
9. The fusion protein of claim 8 , wherein the Fc receptor is an Fcγ receptor.
10. The fusion protein of claim 8 , wherein the Fc receptor is an activating Fc receptor.
11. The fusion protein of claim 8 , wherein the Fc receptor is selected from the group of FcγRIIIa (CD16a), FcγRI (CD64), FcγRIIa (CD32) and FcαRI (CD89).
12. The fusion protein of claim 8 , wherein the Fc receptor is FcγIIIa.
13. The fusion protein of claim 8 , wherein the IgG-class antibody comprises a heavy chain comprising an amino acid substitution at position 329 according to EU numbering.
14. The fusion protein of claim 13 , wherein the amino acid substitution is P329G.
15. The fusion protein claim 8 , wherein the IgG-class antibody comprises a heavy chain comrising amino acid substitutions at positions 234 and 235 according to EU numbering.
16. The fusion protein of claim 15 , wherein the amino acid substitutions are L234A and L235A (LALA).
17. The fusion protein of claim 8 , wherein the IgG-class antibody comprises a heavy chain comrising amino acid substitutions L234A, L235A and P329G according to EU numbering.
18. The fusion protein of claim 1 , wherein the antibody is an IgG-class antibody.
19. The fusion protein of claim 1 , wherein the antibody is IgG 1 -subclass antibody.
20. The fusion protein of claim 1 , wherein the antibody is a full-length antibody.
21. The fusion protein of claim 1 , wherein the antibody is a human antibody.
22. A pharmaceutical composition comprising the fusion protein of claim 1 and a pharmaceutically acceptable carrier.