CD47 antibodies and uses thereof for treating cancer
Humanized antibodies, including masked antibodies that specifically bind to CD47 are provided. Methods for using anti-CD47 antibodies, including masked antibodies, to modulate activity of (e.g., inhibit proliferation of) a CD47-expressing cell, as well as for the treatment of one or more diseases or disorders (e.g., cancer) associated with CD47-expressing cells, are provided.
1. A humanized antibody or antigen-binding fragment thereof that specifically binds human CD47, the antibody or antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, wherein the heavy chain variable region comprises HCDR1 selected from SEQ ID NOs: 16, 19, 21, and 23; HCDR2 selected from SEQ ID NOs: 17, 20, 22, and 24; and HCDR3 of SEQ ID NO: 18; wherein the light chain variable region comprises LCDR1 selected from SEQ ID NOs: 31 and 34; LCDR2 selected from SEQ ID NOs: 32 and 35; and LCDR3 selected from SEQ ID NOs: 33 and 36; wherein the heavy chain variable region comprises an amino acid sequence with at least 97% identity to an amino acid sequence selected from SEQ ID NOs: 3, 4, 5, 6, 7 and 8; and wherein the light chain variable region comprises an amino acid sequence with at least 85% identity to an amino acid sequence selected from SEQ ID NOs: 11, 12, 13, 14 and 15.
2. The humanized antibody or antigen-binding fragment thereof of claim 1 , wherein the heavy chain variable region comprises HCDR1, HCDR2, and HCDR3 selected from: SEQ ID NOs: 16, 17, and 18; SEQ ID NOs: 19, 20, and 18; SEQ ID NOs: 21, 22, and 18; SEQ ID NOs: 16, 20, and 18; and SEQ ID NOs: 23, 24, and 18.
3. The humanized antibody or antigen-binding fragment thereof of claim 1 , wherein the light chain variable region comprises LCDR1, LCDR2, and LCDR3 selected from SEQ ID NOs: 31, 32, and 33; SEQ ID NOs: 31, 32, and 36; and SEQ ID NOs: 34, 35, and 33.
4. The humanized antibody or antigen-binding fragment thereof of claim 1 , wherein the antibody or antigen-binding fragment thereof comprises HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 selected from SEQ ID NOs: 16, 17, 18, 31, 32, and 33; SEQ ID NOs: 16, 17, 18, 34, 35, and 33; SEQ ID NOs: 19, 20, 18, 31, 32, and 33; SEQ ID NOs: 19, 20, 18, 34, 35, and 33; SEQ ID NOs: 21, 22, 18, 31, 32, and 33; SEQ ID NOs: 21, 22, 18, 34, 35, and 33; SEQ ID NOs: 16, 20, 18, 31, 32, and 33; SEQ ID NOs: 16, 20, 18, 34, 35, and 33; SEQ ID NOs: 23, 24, 18, 31, 32, and 33; SEQ ID NOs: 23, 24, 18, 34, 35, and 33; SEQ ID NOs: 16, 17, 18, 31, 32, and 36; SEQ ID NOs: 19, 20, 18, 31, 32, and 36; SEQ ID NOs: 21, 22, 18, 31, 32, and 36; 16, 20, 18, 31, 32, and 36; and SEQ ID NOs: 23, 24, 18, 31, 32, and 36.
5. The humanized antibody or antigen-binding fragment thereof of claim 1 , wherein the heavy chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 3, 4, 5, 6, 7 and 8.
6. The humanized antibody or antigen-binding fragment thereof of claim 1 , wherein the light chain variable region comprises an amino acid sequence selected from SEQ ID NOs: 11, 12, 13, 14 and 15.
7. The humanized antibody or antigen-binding fragment thereof of claim 1 , wherein the heavy chain variable region and light chain variable region comprise SEQ ID NOs: 3 and 11; SEQ ID NOs: 3 and 12; SEQ ID NOs: 3 and 13; SEQ ID NOs: 3 and 14; SEQ ID NOs: 4 and 11; SEQ ID NOs: 4 and 12; SEQ ID NOs: 4 and 13; SEQ ID NOs: 4 and 14; SEQ ID NOs: 5 and 11; SEQ ID NOs: 5 and 12; SEQ ID NOs: 5 and 13; SEQ ID NOs: 5 and 14; SEQ ID NOs: 6 and 11; SEQ ID NOs: 6 and 12; SEQ ID NOs: 6 and 13; SEQ ID NOs: 6 and 14; SEQ ID NOs: 7 and 11; SEQ ID NOs: 7 and 12; SEQ ID NOs: 7 and 13; SEQ ID NOs: 7 and 14; SEQ ID NOs: 8 and 11; SEQ ID NOs: 8 and 12; SEQ ID NOs: 8 and 13; SEQ ID NOs: 8 and 14; or SEQ ID NOs: 3 and 15.
8. A humanized antibody or antigen-binding fragment thereof that specifically binds human CD47, the antibody or antigen-binding fragment comprising a light chain variable region and a heavy chain variable region, wherein the heavy chain variable region comprises HCDR1 of SEQ ID NO: 16, HCDR2 of SEQ ID NO: 17, and HCDR3 of SEQ ID NO: 18; and wherein the light chain variable region comprises LCDR1 of SEQ ID NO: 31, LCDR2 of SEQ ID NO: 32, and LCDR3 of SEQ ID NO: 33; and wherein the heavy chain variable region comprises an amino acid sequence with at least 95% identity to SEQ ID NO: 3 and the light chain variable region comprises an amino acid sequence with at least 85% identity to SEQ ID NO: 13; and wherein the antibody has reduced hemagglutination of red blood cells compared to Ab47 comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 2 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 10.
9. The humanized antibody or antigen-binding fragment thereof of claim 8 , wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 3 and the light chain variable region comprises the amino acid sequence of SEQ ID NO: 13.
10. A nucleic acid sequence encoding the antibody or antigen-binding fragment thereof of claim 1 .
11. An expression vector comprising the nucleic acid of claim 10 .
12. A host cell comprising the nucleic acid of claim 10 .
13. A host cell that expresses the antibody or antigen-binding fragment thereof of claim 1 .
14. A method of producing an antibody or antigen-binding fragment thereof comprising culturing the host cell of claim 12 .
15. A method of producing an antibody or antigen-binding fragment thereof comprising culturing the host cell of claim 13 .
16. A nucleic acid sequence encoding the antibody or antigen-binding fragment thereof of claim 4 .
17. An expression vector comprising the nucleic acid of claim 16 .
18. A host cell comprising the nucleic acid of claim 16 .
19. A host cell that expresses the antibody or antigen-binding fragment thereof of claim 4 .
20. A method of producing an antibody or antigen-binding fragment thereof comprising culturing the host cell of claim 18 .
21. A method of producing an antibody or antigen-binding fragment thereof comprising culturing the host cell of claim 19 .
22. A nucleic acid sequence encoding the antibody or antigen-binding fragment thereof of claim 8 .
23. An expression vector comprising the nucleic acid of claim 22 .
24. A host cell comprising the nucleic acid of claim 22 .
25. A host cell that expresses the antibody or antigen-binding fragment thereof of claim 8 .
26. A method of producing an antibody or antigen-binding fragment thereof comprising culturing the host cell of claim 24 .
27. A method of producing an antibody or antigen-binding fragment thereof comprising culturing the host cell of claim 25 .
28. A nucleic acid sequence encoding the antibody or antigen-binding fragment thereof of claim 9 .
29. An expression vector comprising the nucleic acid of claim 28 .
30. A host cell comprising the nucleic acid of claim 29 .
31. A host cell that expresses the antibody or antigen-binding fragment thereof of claim 9 .
32. A method of producing an antibody or antigen-binding fragment thereof comprising culturing the host cell of claim 30 .
33. A method of producing an antibody or antigen-binding fragment thereof comprising culturing the host cell of claim 31 .