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Monitoring Binding of HIV-1 Capsid Assembly Inhibitors Using 19F Ligand-and 15N Protein-Based NMR and X-ray Crystallography: Early Hit Validation of a Benzodiazepine Series
The emergence of resistance to existing classes of antiretroviral drugs underlines the need to find novel human immunodeficiency virus (HIV)‐1 targets for drug discovery. The viral capsid protein (CA) represents one such potential target. Recently, a series of benzodiazepine inhibitors was identifie...
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Published in: | ChemMedChem 2013-03, Vol.8 (3), p.405-414 |
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Main Authors: | , , , , , , , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Online Access: | Get full text |
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Summary: | The emergence of resistance to existing classes of antiretroviral drugs underlines the need to find novel human immunodeficiency virus (HIV)‐1 targets for drug discovery. The viral capsid protein (CA) represents one such potential target. Recently, a series of benzodiazepine inhibitors was identified via high‐throughput screening using an in vitro capsid assembly assay (CAA). Here, we demonstrate how a combination of NMR and X‐ray co‐crystallography allowed for the rapid characterization of the early hits from this inhibitor series. Ligand‐based 19F NMR was used to confirm inhibitor binding specificity and reversibility as well as to identify the N‐terminal domain of the capsid (CANTD) as its molecular target. Protein‐based NMR (1H and 15N chemical shift perturbation analysis) identified key residues within the CANTD involved in inhibitor binding, while X‐ray co‐crystallography confirmed the inhibitor binding site and its binding mode. Based on these results, two conformationally restricted cyclic inhibitors were designed to further validate the possible binding modes. These studies were crucial to early hit confirmation and subsequent lead optimization.
CA all the way! Early hits from a series of benzodiazepine inhibitors of viral capsid protein (CA), a novel target against HIV, were characterized using NMR and X‐ray co‐crystallography. Ligand‐based 19F NMR was used to confirm binding specificity and reversibility, and to identify the N‐terminal domain (CANTD) as the molecular target. Protein‐based NMR identified key residues involved in binding, while X‐ray co‐crystallography confirmed the binding site and mode. Conformationally restricted cyclic inhibitors further validated the possible binding modes. |
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ISSN: | 1860-7179 1860-7187 |
DOI: | 10.1002/cmdc.201200580 |