Humanized anti-CD70 binding agents and uses thereof
View Patent ↗Disclosed are CD70 binding agents, such as humanized anti-CD70 antibodies and fragments and derivatives, that exert a cytotoxic, cytostatic or immunomodulatory on CD70 expressing cells, as well as pharmaceutical compositions and kits comprising the antibody, fragment or derivative. Also disclosed are methods for the treatment of CD70-expressing cancers and immunological disorders, comprising administering to a subject the CD70 binding agents or pharmaceutical compositions.
1. An isolated polynucleotide encoding a humanized heavy chain variable region comprising the three CDRs from SEQ ID NO:2 and a variable region framework sequence of human germline V H 1-2 or V H 1-18 and exon J H -6, provided that any of positions H46, H67, H68, H69, H70, H71, H80, H81, H82, H82A and H91 (Kabat numbering) can be occupied by the amino acid occupying the corresponding position from SEQ ID NO:2.
2. The isolated polynucleotide of claim 1 , further encoding a human IgG constant region.
3. The isolated polynucleotide of claim 2 , wherein the IgG constant region is IgG1 or IgG3.
4. The isolated polynucleotide of claim 2 , wherein the humanized heavy chain variable region and the human IgG constant region together comprise the amino acid sequence corresponding to residues 20-467 of SEQ ID NO:16 or residues 20-467 of SEQ ID NO:20.
5. The isolated polynucleotide of claim 4 , comprising the nucleotide sequence of nucleotides 58-1401 of SEQ ID NO:15 or nucleotides 58-1401 of SEQ ID NO:19.
6. The isolated polynucleotide of claim 1 , wherein position H46 is occupied by the amino acid occupying the corresponding position from SEQ ID NO:2.
7. The isolated polynucleotide of claim 1 , wherein at least one of positions H46, H67, H68, H69, H70, H71, H80, H81, H82, H82A and H91 (Kabat numbering) in the humanized heavy chain variable region is occupied by the residue occupying the corresponding position in SEQ ID NO:2.
8. The isolated polynucleotide of claim 7 , wherein at least one of positions H71 and H91 (Kabat numbering) in the humanized heavy chain variable region is occupied by the residue occupying the corresponding position in SEQ ID NO:2.
9. The isolated polynucleotide of claim 1 , wherein the humanized heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO:6, SEQ ID NO:10, SEQ ID NO:14, SEQ ID NO:18, or amino acids 20-137 of SEQ ID NO:4.
10. An isolated polynucleotide, comprising the nucleotide sequence set forth in SEQ ID NO:5, SEQ ID NO:9, SEQ ID NO:13, SEQ ID NO:17, or nucleotides 58-411 of SEQ ID NO:3.
11. An isolated polynucleotide encoding a humanized light chain variable region comprising the three CDRs from SEQ ID NO:22 and a variable region framework sequence of human germline Vκ exon B3 and Jκ exon Jκ−1, provided that any of positions L25 and L33 can be occupied by the amino acid occupying the corresponding position from SEQ ID NO:22.
12. The isolated polynucleotide of claim 11 , further encoding a light chain constant region.
13. The isolated polynucleotide of claim 12 , wherein the light chain constant region is a human kappa constant domain.
14. The isolated polynucleotide of claim 12 , wherein the humanized light chain variable region and the light chain constant region together comprise the amino acid sequence corresponding to residues 21-238 of SEQ ID NO:26.
15. The isolated polynucleotide of claim 14 , comprising the nucleotide sequence of nucleotides 61-714 of SEQ ID NO:25.
16. The isolated polynucleotide of claim 11 , wherein at least one of positions L25 and L33 in the humanized light chain variable region is occupied by the residue occupying the corresponding position in SEQ ID NO:22.
17. The isolated polynucleotide of claim 11 , wherein the humanized light chain variable region comprises the amino acid sequence of SEQ ID NO:24.
18. An isolated polynucleotide, comprising the nucleotide sequence set forth in SEQ ID NO:23.
19. The isolated polynucleotide of claim 11 , further encoding a humanized heavy chain variable region comprising the three CDRs from SEQ ID NO:2 and a variable region framework sequence of human germline V H 1-2 or V H 1-18 and exon J H -6, provided that any of positions H46, H67, H68, H69, H70, H71, H80, H81, H82, H82A and H91 (Kabat numbering) can be occupied by the amino acid occupying the corresponding position from SEQ ID NO:2.
20. The isolated polynucleotide of claim 10 , further comprising the nucleotide sequence set forth in SEQ ID NO:23.
21. The isolated polynucleotide of claim 20 , wherein the humanized heavy chain variable region is encoded by the nucleotide sequence set forth in SEQ ID NO:13 or SEQ ID NO:17, and the humanized light chain variable region is encoded by the sequence set forth in SEQ ID NO:23.
22. A vector comprising the isolated polynucleotide of claim 1 .
23. A vector comprising the isolated polynucleotide of claim 11 .
24. A host cell transfected with the vector of claim 22 .
25. A host cell transfected with the vector of claim 23 .
26. The host cell of claim 24 that is a mammalian host cell.
27. The host cell of claim 26 , wherein the mammalian host cell is selected from the group consisting of CHO, VERO, BHK, HeLa, COS, MDCK, 293, 3T3, and W138 cells.
28. The host cell of claim 25 that is a mammalian host cell.
29. The host cell of claim 28 , wherein the mammalian host cell is selected from the group consisting of CHO, VERO, BHK, HeLa, COS, MDCK, 293, 3T3, and W138 cells.
30. A method of producing an antibody, comprising culturing the host cell of claim 24 to produce the antibody.
31. A method of producing an antibody, comprising culturing the host cell of claim 25 to produce the antibody.