Polynucleotides encoding VEGF/DLL4 binding agents
The present invention relates to VEGF-binding agents, DLL4-binding agents, VEGF/DLL4 bispecific binding agents, and methods of using the agents for treating diseases such as cancer. The present invention provides antibodies that specifically bind human VEGF, antibodies that specifically bind human DLL4, and bispecific antibodies that specifically bind human VEGF and/or human DLL4. The present invention further provides methods of using the agents to inhibit tumor growth. Also described are methods of treating cancer comprising administering a therapeutically effect amount of an agent or antibody of the present invention to a patient having a tumor or cancer.
1. An isolated polynucleotide encoding a bispecific antibody that specifically binds human VEGF and human DLL4, wherein the bispecific antibody comprises:
(a) a first heavy chain comprising a heavy chain CDR1 comprising NYWMH (SEQ ID NO:17), a heavy chain CDR2 comprising DINPSNGRTSYKEKFKR (SEQ ID NO:18), and a heavy chain CDR3 comprising HYDDKYYPLMDY (SEQ ID NO:19);
(b) a second heavy chain comprising a heavy chain CDR1 comprising TAYYIH (SEQ ID NO:13) or AYYIH (SEQ ID NO:79), a heavy chain CDR2 comprising YIANYNRATNYNQKFKG (SEQ ID NO:14), YISSYNGATNYNQKFKG (SEQ ID NO:15), YIAGYKDATNYNQKFKG (SEQ ID NO:59), or YISNYNRATNYNQKFKG (SEQ ID NO:65), and a heavy chain CDR3 comprising RDYDYDVGMDY (SEQ ID NO:16); and
(c) a first and a second light chain comprising a light chain CDR1 comprising RASESVDNYGISFMK (SEQ ID NO:20), a light chain CDR2 comprising AASNQGS (SEQ ID NO:21), and a light chain CDR3 comprising QQSKEVPWTFGG (SEQ ID NO:22).
2. The polynucleotide of claim 1 , wherein the second heavy chain comprises a heavy chain CDR1 comprising TAYYIH (SEQ ID NO:13), a heavy chain CDR2 comprising YISNYNRATNYNQKFKG (SEQ ID NO:65), and a heavy chain CDR3 comprising RDYDYDVGMDY (SEQ ID NO:16).
3. The polynucleotide of claim 1 , wherein
(a) the first heavy chain comprises a heavy chain variable region comprising SEQ ID NO:11;
(b) the second heavy chain comprises a heavy chain variable region comprising SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:58, or SEQ ID NO:64; and
(c) the first and second light chains comprise a light chain variable region comprising SEQ ID NO:12.
4. The polynucleotide of claim 3 , wherein the second heavy chain comprises a heavy chain variable region comprising SEQ ID NO:64.
5. The polynucleotide of claim 1 , wherein:
(a) the first heavy chain comprises SEQ ID NO:7;
(b) the second heavy chain comprises SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:56, or SEQ ID NO:62; and
(c) the first and second light chains comprise SEQ ID NO:8.
6. The polynucleotide of claim 5 , wherein the second heavy chain comprises SEQ ID NO:62.
7. A vector comprising the polynucleotide of claim 1 .
8. An isolated cell comprising the polynucleotide of claim 1 .
9. A composition comprising one or more polynucleotides that encode a bispecific antibody that specifically binds human VEGF and human DLL4, wherein the bispecific antibody comprises a first heavy chain of SEQ ID NO:7, a second heavy chain of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:56, or SEQ ID NO:62, and two light chains of SEQ ID NO:8.
10. The composition of claim 9 , wherein a first polynucleotide encodes the first heavy chain of SEQ ID NO:7; a second polynucleotide encodes the second heavy chain of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:56, or SEQ ID NO:62; and a third polynucleotide encodes the light chains of SEQ ID NO:8.
11. The composition of claim 9 , wherein a first polynucleotide encodes the first heavy chain of SEQ ID NO:7 and the light chains of SEQ ID NO:8, and a second polynucleotide encodes the second heavy chain of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:56, or SEQ ID NO:62.
12. The composition of claim 9 , wherein a first polynucleotide encodes the first heavy chain of SEQ ID NO:7, and a second polynucleotide encodes the second heavy chain of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:56, or SEQ ID NO:62 and the light chains of SEQ ID NO:8.
13. The composition of claim 9 , wherein a polynucleotide encodes the first heavy chain of SEQ ID NO:7, the second heavy chain of SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:56, or SEQ ID NO:62 and the light chain of SEQ ID NO:8.
14. An isolated polynucleotide comprising a sequence selected from the group consisting of: SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO:77, and SEQ ID NO:78.
15. An isolated polynucleotide encoding the polypeptide encoded by the plasmid deposited with the ATCC as accession number PTA-13236, PTA-13278, or PTA-13235.
16. A vector comprising:
(a) a polynucleotide encoding a polypeptide comprising SEQ ID NO:7;
(b) a polynucleotide encoding a polypeptide comprising SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:56, or SEQ ID NO:62;
(c) a polynucleotide encoding a first polypeptide comprising SEQ ID NO:7 and a second polypeptide comprising SEQ ID NO:8;
(d) a polynucleotide encoding a first polypeptide comprising SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:56, or SEQ ID NO:62; and a second polypeptide comprising SEQ ID NO:8;
(e) a polynucleotide encoding a first polypeptide comprising SEQ ID NO:7 and a second polypeptide comprising SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:56, or SEQ ID NO:62; or
(f) a polynucleotide encoding a first polypeptide comprising SEQ ID NO:7; a second polypeptide comprising SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:56, or SEQ ID NO:62; and a third polypeptide comprising SEQ ID NO:8.
17. An isolated cell comprising a vector of claim 16 .