Constructs having a SIRP-α 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) polypeptide or a fragment thereof that is capable of binding CD47; and
(b) an Fc variant, wherein the Fc variant is a human IgG1 Fc region comprising mutations L234A, L235A, G237A, and N297A, wherein numbering is according to the EU index,
wherein the polypeptide does not cause acute anemia in rodents, non-human primates, or humans after administration.
2. The polypeptide of claim 1 , wherein the Fc variant binds to an Fcγ receptor with a K D greater than about 5×10 −6 M.
3. The polypeptide of claim 1 , wherein the SIRP-α polypeptide or fragment thereof that is capable of binding CD47 comprises a SIRP-α D1 variant comprising the amino acid sequence, EEELQX 1 IQPDKSVLVAAGETATLRCTX 2 TSLX 3 PVGPIQWFRGAGPGRX 4 LIYNQX 5 EGX 6 FPR VTTVSDX 7 TKRNNMDFSIRIGX 8 ITPADAGTYYCX 9 KFRKGSPDDVEFKSGAGTELSVRAKPS (SEQ ID NO: 51), wherein X 1 is V or I; X 2 is A or I; X 3 is I or F; X 4 is E or V; X 5 is K or R; X 6 is H or P; X 7 is L or T; X 8 is any amino acid other than N; and X 9 is V or I.
4. The polypeptide of claim 3 , wherein the SIRP-α polypeptide or fragment thereof that is capable of binding CD47 comprises a SIRP-α D1 variant wherein X 1 is V or I; X 2 is A or I; X 3 is I or F; X 4 is E; X 5 is K or R; X 6 is H or P; X 7 is L or T; X 8 is not N; and X 9 is V.
5. A method of treating an individual having a disease or disorder, the method comprising administering to the individual a polypeptide comprising:
(a) a signal-regulatory protein α (SIRP-α) polypeptide or a fragment thereof that is capable of binding CD47; and
(b) an Fc variant, wherein the Fc variant a human IgG1 Fc region comprising mutations L234A, L235A, G237A, and N297A,
wherein numbering is according to the EU index;
wherein the polypeptide does not cause acute anemia in the individual after administration.
6. The method of claim 5 , wherein the disease or disorder is a cancer, an autoimmune disease, or an inflammatory disease.
7. The method of claim 5 , wherein the disease or disorder is a cancer, and the cancer is selected from solid tumor cancer, hematological cancer, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, non-Hodgkin lymphoma, Hodgkin lymphoma, multiple myeloma, bladder cancer, pancreatic cancer, cervical cancer, endometrial cancer, lung cancer, bronchus cancer, liver cancer, ovarian cancer, colon and rectal cancer, stomach cancer, gastric cancer, gallbladder cancer, gastrointestinal stromal tumor cancer, thyroid cancer, head and neck cancer, oropharyngeal cancer, esophageal cancer, melanoma, non-melanoma skin cancer, Merkel cell carcinoma, virally induced cancer, neuroblastoma, breast cancer, prostate cancer, renal cancer, renal cell cancer, renal pelvis cancer, leukemia, lymphoma, sarcoma, glioma, brain tumor, and carcinoma.
8. The method of claim 5 , wherein the disease or disorder is an autoimmune disease or an inflammatory disease, and the autoimmune disease or the inflammatory disease is selected from multiple sclerosis, rheumatoid arthritis, a spondyloarthropathy, systemic lupus erythematosus, an antibody-mediated inflammatory or autoimmune disease, graft versus host disease, sepsis, diabetes, psoriasis, atherosclerosis, Sjogren's syndrome, progressive systemic sclerosis, scleroderma, acute coronary syndrome, ischemic reperfusion, Crohn's Disease, endometriosis, glomerulonephritis, myasthenia gravis, idiopathic pulmonary fibrosis, asthma, acute respiratory distress syndrome (ARDS), vasculitis, and inflammatory autoimmune myositis.
9. The method of claim 5 , wherein the signal-regulatory protein α (SIRP-α) polypeptide or fragment thereof that is capable of binding CD47 comprises a sequence set forth in any one of SEQ ID NOs: 78-85.
10. The method of claim 9 , further comprising administration of at least one additional agent.
11. The method of claim 10 , wherein the at least one additional agent is an antibody, tumor associated antigen, or a non-antibody therapeutic.
12. The method of claim 11 , wherein at least two additional agents are administered.
13. The method of claim 12 , wherein the at least two additional agents comprise two antibodies.
14. The method of claim 12 , wherein the at least two additional agents comprise an antibody and a tumor associated antigen.
15. The method of claim 11 , wherein the antibody is a human IgG1 isotype antibody, a human IgG2 isotype antibody, or a human IgG4 isotype antibody.
16. The method of claim 11 , wherein the antibody is selected from an anti-HER2 antibody, anti-CD20 antibody, anti-CD19 antibody, anti-CS1 antibody, anti-CD38 antibody, anti-EGFR antibody, anti-OX40 antibody, anti-PD-1 antibody, anti-PD-L1 antibody, anti-RANKL antibody, anti-CD274 antibody, anti-CTLA-4 antibody, anti-CD137 antibody, anti-4-1BB antibody, anti-B7-H3 antibody, anti-FZD7 antibody, anti-CD27 antibody, anti-CCR4 antibody, anti-CSF1R antibody, anti-CSF antibody, anti-CD30 antibody, anti-BAFF antibody, anti-VEGF antibody, or anti-VEGFR2 antibody.
17. The method of claim 10 , wherein the at least one additional agent is a tumor associated antigen and the tumor associated antigen elicits an immune response.
18. The method of claim 10 , wherein the at least one additional agent is an antibody and the antibody targets a HLA/peptide or MHC/peptide complex.
19. The method of claim 18 , wherein the antibody targets a HLA/peptide or MHC/peptide complex comprising NY-ESO-1/LAGE1, SSX-2, MAGE family (MAGE-A3), gp100/pmel17, Melan-A/MART-1, gp75/TRP1, tyrosinase, TRP2, CEA, PSA, TAG-72, Immature laminin receptor, MOK/RAGE-1, WT-1, Her2/neu, EphA3, SAP-1, BING-4, Ep-CAM, MUC1, PRAME, survivin, Mesothelin, BRCA1/2, CDK4, CML66, MART-2, p53, Ras, β-catenin, HPV E6, or HPV E7.
20. A nucleic acid encoding a polypeptide comprising
(a) a signal-regulatory protein α (SIRP-α) polypeptide or a fragment thereof that is capable of binding CD47; and
(b) an Fc variant wherein the Fc variant is a human IgG1 Fc region comprising mutations L234A, L235A, G237A, and N297A,
wherein numbering is according to the EU index.
21. A vector comprising the nucleic acid of claim 20 .
22. A host cell comprising the nucleic acid of claim 20 .
23. A method of producing a polypeptide comprising culturing the host cell of claim 22 under appropriate conditions to cause expression of the polypeptide and recovering the polypeptide.
24. A pharmaceutical composition comprising:
(I) a polypeptide that comprises
(a) a signal-regulatory protein α (SIRP-α) polypeptide or a fragment thereof that is capable of binding CD47; and
(b) an Fc variant, wherein the Fc variant is a human IgG1 Fc region comprising mutations L234A, L235A, G237A, and N297A,
wherein numbering is according to the EU index; and
(II) a pharmaceutically acceptable carrier.
25. A dimer comprising a polypeptide that comprises:
(a) a signal-regulatory protein α (SIRP-α) polypeptide or a fragment thereof that is capable of binding CD47; and
(b) an Fc variant, wherein the Fc variant is a human IgG1 Fc region comprising mutations L234A, L235A, G237A, and N297A,
wherein numbering is according to the EU index.
26. The dimer of claim 25 , wherein the dimer is a heterodimer.
27. The dimer of claim 25 , wherein the dimer is a homodimer.
28. The polypeptide of claim 1 , wherein the SIRP-α polypeptide or a fragment thereof binds to CD47 with a K D of about 10 nM or less.