Fusion proteins for treating cancer and related methods
Aspects of the disclosure provide fusion proteins that bind cells expressing one or more target molecules including, for example, one or more cell surface multisubunit signaling receptors (e.g., EGFRvIII-expressing cells that also express interferon receptors) and that induce anti-proliferative effects, and related compositions and methods for the treatment of cancer.
1. A fusion protein comprising
a Type 1 interferon protein domain comprising the sequence of SEQ ID NO: 18 or a sequence with one or two mutations relative to SEQ ID NO:18, the mutations selected from the group consisting of:
L30A, R145A, M149A, E59A, H58A, and R150A;
an antibody variable region element comprising a heavy chain variable region comprising the amino acid sequence provided by SEQ ID NO: 15 and a light chain variable region comprising the amino acid sequence provided by SEQ ID NO: 16, wherein the antibody variable region element binds to EGFRvIII; and
a linker that connects the protein domain and the antibody variable region element.
2. The fusion protein of claim 1 , wherein the fusion protein comprises one polypeptide chain.
3. The fusion protein of claim 1 , wherein the Type 1 interferon protein domain comprises mutations H58A and R150A.
4. The fusion protein of claim 1 , wherein the Type 1 interferon comprises mutations E59A and M149A.
5. The fusion protein of claim 1 , wherein the linker connects the C-terminal end of the antibody variable region element to the N-terminal end of the protein domain.
6. The fusion protein of claim 1 , wherein the linker connects the C-terminal end of the protein domain to the N-terminal end of the antibody or antibody variable region element.
7. The fusion protein of claim 1 , wherein the linker is a peptide linker and has a net charge.
8. The fusion protein of claim 1 , wherein the net charge of the linker is negative.
9. The fusion protein of claim 1 , wherein the net charge of the linker is positive.
10. The fusion protein of claim 8 , wherein the linker comprises amino acids selected from the group consisting of glycine, serine, glutamate, and aspartate.
11. The fusion protein of claim 9 , wherein the linker comprises amino acids selected from the group consisting of lysine, arginine, and histidine.
12. The fusion protein of claim 1 , comprising the amino acid sequence of SEQ ID NO: 1 [MR1-1 IFNα2-lb WT].
13. The fusion protein of claim 1 , comprising the amino acid sequence of SEQ ID NO: 2 [MR1-1 IFNα2-1B L30A].
14. The fusion protein of claim 1 , comprising the amino acid sequence of SEQ ID NO: 3 [MR1-1 IFNα2-1B R145A].
15. The fusion protein of claim 1 , comprising the amino acid sequence of SEQ ID NO: 4 [MR1-1 IFNα2-lb E59A M149A].
16. An isolated fusion protein comprising the amino acid sequence of SEQ ID NO: 5 [MR1-1 IFNα2-lb H58A R150A].