Abstract
Peptides are highly selective, high-affinity ligands for a diverse array of disease targets, but suitably derivatizing them for application as diagnostic or therapeutic agents often presents a significant challenge. Covalent modification with metal chelates frequently results in decreased binding affinity, so a variety of strategies must be explored to find suitable locations for modification and facile peptide conjugation chemistries that maintain or enhance binding affinity. In this chapter, we present a paradigm for systematically optimizing peptide binding and determining the favorable sites and methods for peptide conjugation. This strategy is illustrated by two case studies of peptide-based targeted gadolinium contrast agents: EP-2104R for diagnosis of thrombosis and EP-3533 for diagnosis of cardiac perfusion and fibrosis. Two different architectures for the peptide–metal complex conjugation were designed: EP-2104R contains a total of four gadolinium (Gd) chelates linked at the N- and C-termini, whereas EP-3533 is derivatized with three Gd chelates, two on the N-terminus and one on a lysine side chain. Detailed protocols are provided for two Gd chelate conjugation methods.
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Acknowledgements
We would like to thank Thomas McMurry and Phil Graham for their leadership and scientific contributions to the development of EP-2104R and EP-3533. We also acknowledge the many contributions of colleagues at EPIX Pharmaceuticals to the development of these strategies and protocols, including John Amedio, Jaclyn Chasse, Biplab Das, Qing Deng, Stephane Dumas, Matthew Greenfield, Steffi Koerner, Richard Looby, Shrikumar Nair, Luhua Shen, Wei-Chuan Sun, and Stephan Zech. Discovery of the peptide leads was enabled by collaborations with David Buckler, Bob Ladner, Dan Sexton, and Charles Wescott at DYAX Corporation.
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Kolodziej, A.F., Zhang, Z., Overoye-Chan, K., Jacques, V., Caravan, P. (2014). Peptide Optimization and Conjugation Strategies in the Development of Molecularly Targeted Magnetic Resonance Imaging Contrast Agents. In: Nixon, A. (eds) Therapeutic Peptides. Methods in Molecular Biology, vol 1088. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-673-3_13
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DOI: https://doi.org/10.1007/978-1-62703-673-3_13
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