Claims for Patent: 8,562,986

Summary for Patent: 8,562,986

Title:	Engineered anti-alpha V-integrin hybrid antibodies
Abstract:	The invention relates to engineered antibodies which specifically bind to integrin receptors, especially the alpha V integrin receptor subunit. The antibodies comprise the antigen binding sites (CDRs) of a known mouse anti-integrin antibody, as well as hybrid light chain variable sequences, mutated heavy chain variable sequences (Frs) and modified heavy chain constant sequences. The novel antibodies have improved immunogenic and expression properties and elicit excellent anti-angiogenic as well as anti-tumor activities in humans in monotherapy but also and above all in combination with other angiogenesis and tumor inhibiting agents.
Inventor(s):	Goodman; Simon (Griesheim, DE), Hahn; Diane (Seeheim-Jugenheim, DE), Mitjans; Francesc (Igualada, ES), Adan; Jaume (Mataro, ES), Lo; Kin-Ming (Lexington, MA)
Assignee:	Merck Patent GmbH (Darmstadt, DE)
Application Number:	12/669,408
Patent Claims:	1. An engineered recombinant anti-.alpha.v integrin antibody comprising: (i) the light chain CDR regions: CDR1: RASQDISNYLA (SEQ ID No. 7), CDR2: YTSKIHS (SEQ ID No. 8), and CDR3: QQGNTFPYT (SEQ ID No. 9); (ii) the heavy chain CDR regions: CDR1: SFWMH (SEQ ID No. 10), CDR2: YINPRSGYTEYNEIFRD (SEQ ID No. 11), and CDR3: FLGRGAMDY (SEQ ID No. 12); (iii) the light chain framework regions: FR-1: DIQMTQSPSSLSASVGDRVTITC (SEQ ID No. 14), FR-2: WYQQKPGKAPKLLIY (SEQ ID No. 15), FR-3: GVPSRFSGSGSGTDYTFTISSLQPEDIATYYC (SEQ ID No. 16), and FR-4: FGQGTKVEIK (SEQ ID No. 17) (iv) the heavy chain framework regions: FR1: QVQLQQSGGELAKPGASVKVSCKASGYTFS (SEQ ID No. 22), FR2: WVRQAPGQGLEWIG (SEQ ID No. 23), FR3: KATMTTDTSTSTAYMELSSLRSEDTAVYYCAS (SEQ ID No. 24), and FR4: WGQGTTVTVSS (SEQ ID No. 25); and (v) a heavy chain constant region derived from human IgG and a human constant light chain region. 2. The engineered antibody of claim 1, wherein the heavy chain constant region is derived from IgG2. 3. The engineered antibody of claim 2, wherein said IgG2 constant region comprises a modified IgG1 hinge region. 4. The engineered antibody of claim 3, wherein said modified IgG1 hinge region comprises the sequence EPKSSDKTHTCPPCP (SEQ ID No. 26). 5. The engineered antibody of claim 2, wherein said IgG2 constant region is modified by replacing amino acid N with Q at position 297 (N297Q). 6. The engineered antibody of claim 5, wherein amino acid residue F at position 296 is replaced by A (F296A) in order to eliminate a T-cell epitope generated by the modification at position 297. 7. The engineered antibody of claim 1, wherein the light chain constant region is human kappa. 8. The recombinant anti-.alpha.v-integrin hybrid antibody of claim 1 comprising: (i) variable and constant light chain sequences: DIQMTQSPSSLSASVGDRVTITCRASQDISNYLAWYQQKPGKAPKLLIYYTSKIHSGVPS RFSGSGSGTDYTFTISSLQPEDIATYYCQQGNTFPYTFGQGTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID No. 5) and (ii) variable and constant heavy chain sequences: QVQLQQSGGELAKPGASVKVSCKASGYTFSSFWMHWVRQAPGQGLEWIGYINPRSGY TEYNEIFRDKATMTTDTSTSTAYMELSSLRSEDTAVYYCASFLGRGAMDYWGQGTTVT VSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVEPKSSDKTHTCPPCPAPPV AGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREE QAQSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPP SREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID No. 6). 9. A pharmaceutical composition comprising a pharmaceutically effective amount of the antibody of claim 1 and a pharmaceutically acceptable carrier. 10. A pharmaceutical composition comprising a first and second pharmaceutically effective therapeutic agent, wherein the first agent is the antibody of claim 1, and the second agent is selected from the group consisting of: a chemotherapeutic agent, an angiogenesis inhibitor and an anti-tumor agent. 11. The pharmaceutical composition of claim 10, wherein the anti-tumor agent is an anti-tumor antibody. 12. The antibody of claim 1, wherein the antibody elicits a direct effect on tumor growth, which is independent of the indirect anti-tumor effect of the antibody caused by blocking angiogenesis. 13. The pharmaceutical composition of claim 11, wherein the anti-tumor antibody is an anti-EGFR antibody or an anti-Her2 antibody. 14. The pharmaceutical composition of claim 13, wherein the anti-EGFR antibody is cetuximab or matuzumab. 15. The pharmaceutical composition of claim 10, wherein the anti-angiogenesis inhibitor is the integrin inhibitor cilengitide. 16. The pharmaceutical composition of claim 10, wherein the chemotherapeutic agent is cisplatin or DTIC. 17. An engineered recombinant anti-.alpha.v integrin antibody comprising: (i) the light chain CDR regions: CDR1: RASQDISNYLA (SEQ ID No. 7), CDR2: YTSKIHS (SEQ ID No. 8), and CDR3: QQGNTFPYT (SEQ ID No. 9) and (ii) the heavy chain CDR regions: CDR1: SFWMH (SEQ ID No. 10), CDR2: YINPRSGYTEYNEIFRD (SEQ ID No. 11), and CDR3: FLGRGAMDY (SEQ ID No. 12). 18. The engineered antibody of claim 17, further comprising: (iii) the light chain framework regions: FR-1: DIQMTQSPSSLSASVGDRVTITC (SEQ ID No. 14), FR-2: WYQQKPGKAPKLLIY (SEQ ID No. 15), FR-3: GVPSRFSGSGSGTDYTFTISSLQPEDIATYYC (SEQ ID No. 16), and FR-4: FGQGTKVEIK (SEQ ID No. 17). 19. The engineered antibody of claim 17, further comprising the heavy chain framework regions FR1: QVQLQQSGAELAEPGASVKMSCKASGYTFS (SEQ ID No. 18) FR2: WVKQRPGQGLEWIG (SEQ ID No. 19) FR3: KATMTADTSSSTAYMQLSGLTSEDSAVYYCAS (SEQ ID No. 20) FR4: WGQGTSVTVSS (SEQ ID No. 21), wherein one, more or all of the bold and underlined positions are mutated in order to reduce or eliminate T-cell epitopes and thus immunogenicity in a human. 20. The engineered antibody of claim 19, wherein the heavy chain framework regions are: FR1: QVQLQQSGGELAKPGASVKVSCKASGYTFS (SEQ ID No. 22), FR2: WVRQAPGQGLEWIG (SEQ ID No. 23), FR3: KATMTTDTSTSTAYMELSSLRSEDTAVYYCAS (SEQ ID No. 24), and FR4: WGQGTTVTVSS (SEQ ID No. 25). 21. The engineered antibody of claim 17, further comprising a heavy chain constant region deriving from human IgG and a human constant light chain region. 22. The engineered antibody of claim 21, wherein the heavy chain constant region is derived from IgG2. 23. The engineered antibody of claim 22, wherein said IgG2 constant region comprises a modified IgG1 hinge region. 24. The engineered antibody of claim 23, wherein said modified IgG1 hinge region comprises the sequence EPKSSDKTHTCPPCP (SEQ ID No. 26). 25. The engineered antibody of claim 22, wherein said IgG2 constant region is modified by replacing amino acid N with Q at position 297 (N297Q). 26. The engineered antibody of claim 25, wherein amino acid residue F at position 296 is replaced by A (F296A) in order to eliminate a T-cell epitope generated by the modification at position 297. 27. The engineered antibody of claim 17, wherein the light chain constant region is human kappa. 28. The recombinant anti-.alpha.v-integrin hybrid antibody of claim 17 comprising: (i) variable and constant light chain sequences: DIQMTQSPSSLSASVGDRVTITCRASQDISNYLAWYQQKPGKAPKLLIYYTSKIHSGVPS RFSGSGSGTDYTFTISSLQPEDIATYYCQQGNTFPYTFGQGTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID No. 5) and (ii) variable and constant heavy chain sequences: QVQLQQSGGELAKPGASVKVSCKASGYTFSSFWMHWVRQAPGQGLEWIGYINPRSGY TEYNEIFRDKATMTTDTSTSTAYMELSSLRSEDTAVYYCASFLGRGAMDYWGQGTTVT VSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVL QSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVEPKSSDKTHTCPPCPAPPV AGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREE QAQSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPP SREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID No. 6). 29. A pharmaceutical composition comprising a pharmaceutically effective amount of the antibody of claim 17 and a pharmaceutically acceptable carrier. 30. A pharmaceutical composition comprising a first and second pharmaceutically effective therapeutic agent, wherein the first agent is the antibody of claim 17, and the second agent is selected from the group consisting of: a chemotherapeutic agent, an angiogenesis inhibitor and an anti-tumor agent. 31. The pharmaceutical composition of claim 30, wherein the anti-tumor agent is an anti-tumor antibody. 32. The pharmaceutical composition of claim 31, wherein the anti-tumor antibody is an anti-EGFR antibody or an anti-Her2 antibody. 33. The pharmaceutical composition of claim 32, wherein the anti-EGFR antibody is cetuximab or matuzumab. 34. The pharmaceutical composition of claim 30, wherein the anti-angiogenesis inhibitor is the integrin inhibitor cilengitide. 35. The pharmaceutical composition of claim 30, wherein the chemotherapeutic agent is cisplatin or DTIC. 36. The antibody of claim 17, wherein the antibody elicits a direct effect on tumor growth, which is independent of the indirect anti-tumor effect of the antibody caused by blocking angiogenesis.

Details for Patent 8,562,986

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Eli Lilly And Company	ERBITUX	cetuximab	Injection	125084	02/12/2004		2027-07-17
Eli Lilly And Company	ERBITUX	cetuximab	Injection	125084	03/28/2007		2027-07-17