Site-specific targeting of antibody activity in vivo mediated by disease-associated proteases.
As a general strategy to selectively target antibody activity in vivo, a molecular architecture was designed to render binding activity dependent upon proteases in disease tissues. A protease-activated antibody (pro-antibody) targeting vascular cell adhesion molecule 1 (VCAM-1), a marker of atherosclerotic plaques, was constructed by tethering a binding site-masking peptide to the antibody via a matrix metalloprotease (MMP) susceptible linker. Pro-antibody activation in vitro by MMP-1 yielded a 200-fold increase in binding affinity and restored anti-VCAM-1 binding in tissue sections from ApoE⁻/⁻ mice ex vivo. The pro-antibody was efficiently activated by native proteases in aorta tissue extracts from ApoE⁻/⁻, but not from normal mice, and accumulated in aortic plaques in vivo with enhanced selectivity when compared to the unmodified antibody. Pro-antibody accumulation in aortic plaques was MMP-dependent, and significantly inhibited by a broad-spectrum MMP inhibitor. These results demonstrate that the activity of disease-associated proteases can be exploited to site-specifically target antibody activity in vivo.
Department of Chemical Engineering, University of California, Santa Barbara, CA 93106, USA.
Oran
O
Erster
Jerry M
JM
Thomas
Juliana
J
Hamzah
Abeer M
AM
Jabaiah
Jennifer A
JA
Getz
Tobias D
TD
Schoep
Sejal S
SS
Hall
Erkki
E
Ruoslahti
Patrick S
PS
Daugherty
D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage* / Q000378:metabolism / Q000493:pharmacokinetics; D001011:Aorta / Q000378:metabolism; D001057:Apolipoproteins E / Q000172:deficiency / Q000235:genetics; D002460:Cell Line; D003324:Coronary Artery Disease / Q000378:metabolism / Q000473:pathology; D004195:Disease Models, Animal; D016503:Drug Delivery Systems; D057809:HEK293 Cells; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000008:administration & dosage* / Q000378:metabolism / Q000493:pharmacokinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D058226:Plaque, Atherosclerotic / Q000378:metabolism*; D011355:Prodrugs / Q000008:administration & dosage*; D014018:Tissue Distribution; D019010:Vascular Cell Adhesion Molecule-1 / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D001057:Apolipoproteins E; D011355:Prodrugs; D019010:Vascular Cell Adhesion Molecule-1; D020781:Matrix Metalloproteinase 1