Interleukin-2/Interleukin-2 receptor alpha fusion proteins and methods of use
Disclosed herein are fusion proteins comprising: (a) a first polypeptide comprising Interleukin-2 (IL2); and (b) a second polypeptide, fused in frame to the first polypeptide, wherein the second polypeptide comprises an extracellular domain of Interleukin-2 Receptor alpha (IL2Rα), wherein IL2 or IL2Rα comprises at least one fewer glycosylation site compared to native IL2 or native IL2Rα. Methods of production and methods of therapeutic use of the fusion proteins are also disclosed.
1. A fusion protein comprising:
(a) a first polypeptide comprising an Interleukin-2 (IL2) polypeptide, wherein the first polypeptide comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 2; and
(b) a second polypeptide comprising an extracellular domain of an Interleukin-2 Receptor alpha (IL2Rα) polypeptide, wherein the second polypeptide comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 12;
wherein the fusion protein does not include the C-terminal portion corresponding to amino acids 192 to 219 of the extracellular domain of native IL2Rα (SEQ ID NO: 7); and
wherein the fusion protein has IL2 activity.
2. The fusion protein of claim 1 , wherein the first polypeptide comprises an amino acid sequence having at least 96% sequence identity to SEQ ID NO: 2, and wherein the second polypeptide comprises an amino acid sequence having at least 96% sequence identity to SEQ ID NO: 12.
3. The fusion protein of claim 1 , wherein the first polypeptide comprises an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 2, and wherein the second polypeptide comprises an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 12.
4. The fusion protein of claim 1 , wherein the first polypeptide comprises an amino acid sequence having at least 98% sequence identity to SEQ ID NO: 2, and wherein the second polypeptide comprises an amino acid sequence having at least 98% sequence identity to SEQ ID NO: 12.
5. The fusion protein of claim 1 , wherein the first polypeptide comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 2, and wherein the second polypeptide comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 12.
6. The fusion protein of claim 1 , wherein the first polypeptide comprises the amino acid sequence as set forth in SEQ ID NO: 2, and wherein the second polypeptide comprises the amino acid sequence as set forth in SEQ ID NO: 12.
7. The fusion protein of claim 1 , wherein the amino acid sequence in the first polypeptide has one or more substitutions at amino acid T3, amino acid C125, and a combination thereof, wherein the amino acid locations correspond to SEQ ID NO: 2.
8. The fusion protein of claim 1 , wherein the amino acid sequence in the first polypeptide has a deletion at amino acid A1, wherein the amino acid location corresponds to SEQ ID NO: 2.
9. The fusion protein of claim 1 , further comprising a glycine/serine linker fused in frame between the first polypeptide and the second polypeptide.
10. The fusion protein of claim 9 , wherein the glycine/serine liker comprises the amino acid sequence of (GGGS) 3 .
11. The fusion protein of claim 1 , wherein the fusion protein comprises the amino acid sequence as set forth in SEQ ID NO: 16, SEQ ID NO: 20, SEQ ID NO: 34, SEQ ID NOs: 39 to 41, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NOs: 49 to 53, SEQ ID NOs: 57 to 59, or SEQ ID NOs: 67 to 70.
12. The fusion protein of claim 1 , wherein the fusion protein comprises the amino acid sequence as set forth in SEQ ID NO: 16.
13. The fusion protein of claim 1 , wherein the fusion protein is deglycosylated enzymatically or chemically.
14. The fusion protein of claim 1 , wherein the fusion protein further comprises a heterologous moiety fused to the first polypeptide and/or the second polypeptide.
15. The fusion protein of claim 14 , wherein the heterologous moiety is a half-life extending moiety.
16. The fusion protein of claim 14 , wherein the heterologous moiety comprises albumin, an immunoglobulin constant region or a portion thereof, an immunoglobulin-binding polypeptide, an immunoglobulin G (IgG), albumin-binding polypeptide (ABP), a PASylation moiety, a HESylation moiety, XTEN, a PEGylation moiety, an Fc region, or any combination thereof.
17. The fusion protein of claim 1 , which is a monomer or dimer.
18. A monomer comprising the fusion protein of claim 1 .
19. A dimer comprising two fusion proteins, wherein each of the fusion proteins comprises the fusion protein of claim 1 .
20. The dimer of claim 19 , wherein the fusion proteins are associated with each other via a covalent bond.
21. A composition comprising the fusion protein of claim 1 .
22. A pharmaceutical composition comprising (a) the fusion protein of claim 1 ; and (b) a pharmaceutically acceptable excipient.
23. A kit comprising the fusion protein of claim 1 and instructions for administering the fusion protein to a subject in need thereof.