Constructs having a SIRP-alpha domain or variant thereof
The present disclosure features signal-regulatory protein α (SIRP-α) polypeptides and constructs that are useful, e.g., to target a cell (e.g., a cancer cell or a cell of the immune system), to increase phagocytosis of the target cell, to eliminate immune cells such as regulatory T-cells, to kill cancer cells, to treat a disease (e.g., cancer) in a subject, or any combinations thereof. The SIRP-α constructs include a high affinity SIRP-α D1 domain or variant thereof that binds CD47 with higher affinity than a wild-type SIRP-α. The SIRP-α polypeptides or constructs include a SIRP-α D1 variant fused to an Fc domain monomer, a human serum albumin (HSA), an albumin-binding peptide, or a polyethylene glycol (PEG) polymer. Compositions provided herein include (i) a polypeptide including a signal-regulatory protein α (SIRP-α) D1 variant and (ii) an antibody.
1. A polypeptide, comprising:
(a) a signal-regulatory protein α (SIRP-α) D1 variant comprising SEQ ID NO: 80; and
(b) a human IgG1 Fc region comprising L234A, L235A, G237A, and N297A mutations, wherein numbering is according to the EU index,
wherein the C-terminus of the SIRP-αD1 variant is linked to the N-terminus of the human IgG1 Fc region.
2. A nucleic acid encoding the polypeptide of claim 1 .
3. A vector comprising the nucleic acid of claim 2 .
4. A host cell comprising the nucleic acid of claim 2 .
5. A method of producing a polypeptide, comprising culturing the host cell of claim 4 under appropriate conditions to cause expression of the polypeptide and recovering the polypeptide.
6. A pharmaceutical composition comprising the polypeptide of claim 1 .
7. A pharmaceutical composition comprising
(a) a dimer comprising the polypeptide of claim 1 , and
(b) a pharmaceutically acceptable excipient.
8. The pharmaceutical composition of claim 7 , where the dimer is a homodimer.
9. A method of treating an individual having cancer, the method comprising administering to the individual a pharmaceutical composition comprising the polypeptide of claim 1 .
10. The method of claim 9 , further comprising administering at least one additional agent to the individual, wherein the at least one additional agent is an antibody, a tumor associated antigen, or a non-antibody therapeutic.
11. A method of treating an individual having cancer, the method comprising administering to the individual a pharmaceutical composition comprising a dimer and a pharmaceutically acceptable excipient, wherein the dimer comprises the polypeptide of claim 1 .
12. The method of claim 11 , wherein the dimer is a homodimer.
13. The method of claim 11 , further comprising administering at least one additional agent to the individual, wherein the at least one additional agent is an antibody, a tumor associated antigen, or a non-antibody therapeutic.
14. A dimer comprising a polypeptide that comprises the amino acid sequence set forth in SEQ ID NO: 137.
15. The dimer of claim 14 , wherein each monomer in the dimer comprises a polypeptide that comprises the amino acid sequence set forth in SEQ ID NO: 137.
16. A nucleic acid encoding a polypeptide that comprises the amino acid sequence set forth in SEQ ID NO: 137.
17. A vector comprising the nucleic acid of claim 16 .
18. A host cell comprising the nucleic acid of claim 16 .
19. A method of producing a polypeptide, comprising culturing the host cell of claim 18 under appropriate conditions to cause expression of the polypeptide and recovering the polypeptide.
20. A pharmaceutical composition comprising a polypeptide that comprises the amino acid sequence set forth in SEQ ID NO: 137 and a pharmaceutically acceptable excipient.
21. A pharmaceutical composition comprising (a) the dimer of claim 14 and (b) a pharmaceutically acceptable excipient.
22. The pharmaceutical composition of claim 21 , wherein the dimer is a homodimer.
23. A method of treating an individual having cancer, the method comprising administering to the individual the pharmaceutical composition of claim 20 .
24. The method of claim 23 , further comprising administering at least one additional agent to the individual, wherein the at least one additional agent is an antibody, a tumor associated antigen, or a non-antibody therapeutic.
25. A method of treating an individual having cancer, the method comprising administering to the individual the pharmaceutical composition of claim 21 .
26. A method of treating an individual having cancer, the method comprising administering to the individual the pharmaceutical composition of claim 22 .
27. The method of claim 25 , further comprising administering at least one additional agent to the individual, wherein the at least one additional agent is an antibody, a tumor associated antigen, or a non-antibody therapeutic.
28. A polypeptide, comprising:
(a) a signal-regulatory protein α (SIRP-α) D1 variant comprising SEQ ID NO: 85; and
(b) a human IgG2 Fc region comprising A330S, P331S, and N297A mutations, wherein numbering is according to the EU index,
wherein the C-terminus of the SIRP-αD1 variant is linked to the N-terminus of the human IgG1 Fc region.
29. A nucleic acid encoding a polypeptide that comprises the polypeptide of claim 28 .
30. A vector comprising the nucleic acid of claim 29 .
31. A host cell comprising the nucleic acid of claim 29 .
32. A method of producing a polypeptide, comprising culturing the host cell of claim 31 under appropriate conditions to cause expression of the polypeptide and recovering the polypeptide.
33. A pharmaceutical composition comprising the polypeptide of claim 28 .
34. A pharmaceutical composition comprising
(a) a dimer comprising the polypeptide of claim 28 , and
(b) a pharmaceutically acceptable excipient.
35. The pharmaceutical composition of claim 34 , wherein the dimer is a homodimer.
36. A method of treating an individual having cancer, the method comprising administering to the individual the pharmaceutical composition of claim 33 .
37. The method of claim 36 , further comprising administering at least one additional agent to the individual, wherein the at least one additional agent is an antibody, a tumor associated antigen, or a non-antibody therapeutic.
38. A method of treating an individual having cancer, the method comprising administering to the individual a pharmaceutical composition comprising a dimer and a pharmaceutically acceptable excipient, wherein the dimer comprises the polypeptide of claim 28 .
39. The method of claim 38 , wherein the dimer is a homodimer.
40. The method of claim 38 , further comprising administering at least one additional agent to the individual, wherein the at least one additional agent is an antibody, a tumor associated antigen, or a non-antibody therapeutic.
41. A dimer comprising a polypeptide that comprises the amino acid sequence set forth in SEQ ID NO: 113.
42. The dimer of claim 41 , wherein each monomer in the dimer comprises a polypeptide that comprises the amino acid sequence set forth in SEQ ID NO: 113.
43. A nucleic acid encoding a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 113.
44. A vector comprising the nucleic acid of claim 43 .
45. A host cell comprising the nucleic acid of claim 43 .
46. A method of producing a polypeptide, comprising culturing the host cell of claim 45 under appropriate conditions to cause expression of the polypeptide and recovering the polypeptide.
47. A pharmaceutical composition comprising a polypeptide comprising the amino acid sequence set forth in SEQ ID NO: 113 and a pharmaceutically acceptable excipient.
48. A pharmaceutical composition comprising (a) the dimer of claim 41 and (b) a pharmaceutically acceptable excipient.
49. The pharmaceutical composition of claim 48 , wherein the dimer is a homodimer.
50. A method of treating an individual having cancer, the method comprising administering to the individual a pharmaceutical composition comprising a polypeptide that comprises the amino acid sequence of SEQ ID NO: 113 and a pharmaceutically acceptable excipient.
51. The method of claim 50 , further comprising administering at least one additional agent to the individual, wherein the at least one additional agent is an antibody, a tumor associated antigen, or a non-antibody therapeutic.
52. A method of treating an individual having cancer, the method comprising administering to the individual a pharmaceutical composition comprising the dimer of claim 41 and a pharmaceutically acceptable excipient.
53. The method of claim 52 , wherein the dimer is a homodimer.
54. The method of claim 52 , further comprising administering at least one additional agent to the individual, wherein the at least one additional agent is an antibody, a tumor associated antigen, or a non-antibody therapeutic.