CD8 binding agents
The present invention relates, in part, to agents that bind CD8 and their use as therapeutic and diagnostic agents. The present invention further relates to pharmaceutical compositions comprising the CD8 binding agents and their use in the treatment of various diseases, including, for example, cancers.
1. A CD8 binding agent comprising at least one targeting moiety comprising three complementarity determining regions (CDR1, CDR2, and CDR3), wherein:
CDR1 comprises the amino acid sequence of SEQ ID NO: 28; CDR2 comprises the amino acid sequence of SEQ ID NO: 97; CDR3 comprises the amino acid sequence of SEQ ID NO: 166;
CDR1 comprises the amino acid sequence of SEQ ID NO: 45; CDR2 comprises the amino acid sequence of SEQ ID NO: 1221; CDR3 comprises the amino acid sequence of SEQ ID NO: 181;
CDR1 comprises the amino acid sequence of SEQ ID NO: 39; CDR2 comprises the amino acid sequence of SEQ ID NO: 108; CDR3 comprises the amino acid sequence of SEQ ID NO: 177;
CDR1 comprises the amino acid sequence of SEQ ID NO: 37; CDR2 comprises the amino acid sequence of SEQ ID NO: 112; CDR3 comprises the amino acid sequence of SEQ ID NO: 181; or
CDR1 comprises the amino acid sequence of SEQ ID NO: 51; CDR2 comprises the amino acid sequence of SEQ ID NO: 130; CDR3 comprises the amino acid sequence of SEQ ID NO: 199.
2. The CD8 binding agent of claim 1 , wherein the targeting moiety is a single domain antibody, or a recombinant heavy-chain-only antibody (VHH).
3. The CD8 binding agent of claim 1 , wherein the targeting moiety is a single-domain antibody.
4. The CD8 binding agent of claim 1 , wherein the CD8 binding agent comprises one or more signaling agents.
5. The CD8 binding agent of claim 4 , wherein the signaling agent is selected from one or more of an interferon, an interleukin, and a tumor necrosis factor, and a modified form thereof.
6. The CD8 binding agent of claim 5 , wherein the signaling agent is a modified human IFN-α2 having an amino acid sequence that is at least 95% identical to SEQ ID NO: 288 or 289, said modified human IFN-α2 having one or more mutations that confer improved safety as compared to a wild type IFN-α2; and wherein the targeting moiety and the modified signaling agent are optionally connected with one or more linkers.
7. The CD8 binding agent of claim 6 , wherein the modified human IFN-α2 comprises one or more mutations at positions R120, M148, A145, R149, and L153 relative to SEQ ID NO: 288 or 289.
8. The CD8 binding agent of claim 7 , wherein the modified human IFN-α2 comprises one or more mutations selected from R120E, A145G, R149A, and L153A relative to SEQ ID NO: 288 or 289.
9. The CD8 binding agent of claim 6 , wherein the modified human IFN-α2 comprises a R120E mutation and either a A145G, R149A, or a L153A mutation relative to SEQ ID NO: 288 or 289.
10. The CD8 binding agent of claim 5 , wherein the signaling agent is a modified human IFN-β3 having an amino acid sequence that is at least 95% identical to SEQ ID NO: 290, said modified human IFN-β3 having one or more mutations that confer improved safety as compared to a wild type IFN-β3; and wherein the targeting moiety and the modified signaling agent are optionally connected with one or more linkers.
11. The CD8 binding agent of claim 10 , wherein the modified human IFN-β3 comprises one or more mutations at positions W22, R27, L32, R35, V148, L151, R152, and Y155 relative to SEQ ID NO: 290.
12. The CD8 binding agent of claim 11 , wherein the modified human IFN-β3 comprises one or more mutations selected from W22G, R27G, L32A, L32G, R35A, R35G, V148G, L151G, R152A, and R152G relative to SEQ ID NO: 290.
13. The CD8 binding agent of claim 1 , wherein the CD8 binding agent comprises one or more additional targeting moieties.
14. The CD8 binding agent of claim 13 , wherein the one or more additional targeting moieties recognize or functionally modulate a tumor antigen.
15. The CD8 binding agent of claim 1 , wherein the one or more additional targeting moieties recognize or functionally modulate an antigen on an immune cell.
16. The CD8 binding agent of claim 1 , wherein the CD8 binding agent recruits cytotoxic T cells to tumor cells or to the tumor environment.
17. The CD8 binding agent of claim 1 , wherein the CD8 binding agent recognizes and binds CD8 without substantially functionally modulating its activity.
18. A recombinant nucleic acid composition encoding the CD8 binding agents of claim 1 .
19. A host cell comprising a nucleic acid of claim 18 .
20. The CD8 binding agent of claim 1 , comprising an amino acid sequence having at least 90% identity with one of SEQ ID NOs: 1216-1220.
21. The CD8 binding agent of claim 1 , wherein the CD8 binding agent binds to both human and cynomolgus monkey CD8.
22. A method for treating or preventing cancer, comprising administering to a patient in need thereof an effective amount of the CD8 binding agent of claim 2 .