Engineered antibody-interferon mutant fusion molecules
The field of the present invention relates to genetically engineered fusion molecules, methods of making said fusion molecules, and uses thereof in anti-tumor immunotherapies. More specifically, the present invention relates to fusion molecule constructs wherein a tumor associated antigen (TAA) antibody (Ab) serves as a targeting moiety to selectively deliver a cytokine to a tumor cell for purposes of killing or inhibiting the growth or proliferation of said tumor cell. In various embodiments, the engineered fusion molecules comprise a TAA Ab fused to an interferon-alpha (IFN-α) mutant molecule. The engineered Ab-IFN-α mutant fusion molecules of the present invention demonstrate improved therapeutic index and preserved or increased efficacy as compared to Ab-wildtype IFN-α fusion molecules, and/or demonstrate improved PK properties as compared to Ab-wildtype IFN-α fusion molecules.
1. A genetically engineered fusion molecule comprising a tumor-associated antigen (TAA) antibody (Ab) attached to an interferon alpha (IFN-α) mutant molecule, wherein said antibody is attached directly to said IFN-α mutant molecule, wherein said IFN-α mutant molecule is a mutated human IFN-α2 molecule comprising at least one mutation in SEQ ID NO: 13, wherein said mutation is selected from H57A, E58A, L30A, and F27A, and wherein the TAA antibody is an IgG antibody.
2. A fusion molecule of claim 1 , wherein said fusion molecule demonstrates improved pharmacokinetics (PK) properties as compared to a TAA Ab-wildtype_IFN-α fusion molecule.
3. A fusion molecule of claim 1 , wherein said fusion molecule when contacted to a tumor cell results in the killing or inhibition of growth or proliferation of said tumor cell.
4. A fusion molecule of claim 1 , wherein said TAA antibody is an antibody selected from the group consisting of anti-HER2/neu, anti-HER3, anti-HER4, anti-CD4, anti-CD19, anti-CD20, anti-CD22, anti-CD25, anti-CD33, anti-CD138, anti-CD200, anti-CD276, anti-CXCR3, anti-CXCR5, anti-CCR3, anti-CCR4, anti-CCR9, anti-CRTH2, anti-PMCH, and anti-endoplasmin antibody.
5. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-HER2/neu antibody and wherein the mutated human IFN-α2 molecule comprises the mutation F27A in SEQ ID NO: 13.
6. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-CD20 antibody and wherein the mutated human IFN-α2 molecule comprises the mutation F27A in SEQ ID NO: 13.
7. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-CD138 antibody and wherein the mutated human IFN-α2 molecule comprises the mutation F27A in SEQ ID NO: 13.
8. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-endoplasmin antibody and wherein the mutated human IFN-α2 molecule comprises the mutation F27A in SEQ ID NO: 13.
9. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-CD33 antibody and wherein the mutated human IFN-α2 molecule comprises the mutation F27A in SEQ ID NO: 13.
10. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-CD276 antibody and wherein the mutated human IFN-α2 molecule comprises the mutation F27A in SEQ ID NO: 13.
11. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-HER2/neu antibody and wherein the mutated human IFN-α2 molecule comprises the two mutations R149A and R162A in SEQ ID NO: 13.
12. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-CD20 antibody and wherein the mutated human IFN-α2 molecule comprises the two mutations R149A and R162A in SEQ ID NO: 13.
13. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-CD138 antibody and wherein the mutated human IFN-α2 molecule comprises the two mutations R149A and R162A in SEQ ID NO: 13.
14. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-endoplasmin antibody and wherein the mutated human IFN-α2 molecule comprises the two mutations R149A and R162A in SEQ ID NO: 13.
15. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-CD33 antibody and wherein the mutated human IFN-α2 molecule comprises the two mutations R149A and R162A in SEQ ID NO: 13.
16. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-CD276 antibody and wherein the mutated human IFN-α2 molecule comprises the two mutations R149A and R162A in SEQ ID NO: 13.
17. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-HER2/neu antibody and wherein the mutated human IFN-α2 molecule comprises the mutation R162A in SEQ ID NO: 13.
18. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-CD20 antibody and wherein the mutated human IFN-α2 molecule comprises the mutation R162A in SEQ ID NO: 13.
19. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-CD138 antibody and wherein the mutated human IFN-α2 molecule comprises the mutation R162A in SEQ ID NO: 13.
20. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-endoplasmin antibody and wherein the mutated human IFN-α2 molecule comprises the mutation R162A in SEQ ID NO: 13.
21. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-CD33 antibody and wherein the mutated human IFN-α2 molecule comprises the mutation R162A in SEQ ID NO: 13.
22. A fusion molecule of claim 1 , wherein the TAA antibody is an anti-CD276 antibody and wherein the mutated human IFN-α2 molecule comprises the mutation R162A in SEQ ID NO: 13.
23. A fusion molecule of claim 1 , wherein said antibody is directly joined attached to said interferon mutant molecule with a peptide linker, wherein said peptide linker is selected from SEQ ID NO: 14 and SEQ ID NO: 15.
24. A pharmaceutical composition comprising a fusion molecule of claim 1 in a pharmaceutically acceptable carrier.