Factors affecting irreversible inhibition of EGFR and influence of chirality on covalent binding

Abstract The discovery of targeted covalent inhibitors is of increasing importance in drug discovery. Finding efficient covalent binders requires modulation of warhead reactivity and optimisation of warhead geometry and non-covalent interactions. Uncoupling the contributions that these factors make...

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Main Authors: Pasquale A. Morese, Ayaz Ahmad, Mathew P. Martin, Richard A. Noble, Sara Pintar, Lan Z. Wang, Shangze Xu, Andrew Lister, Richard A. Ward, Agnieszka K. Bronowska, Martin E. M. Noble, Hannah L. Stewart, Michael J. Waring
Format: Article
Language:English
Published: Nature Portfolio 2025-04-01
Series:Communications Chemistry
Online Access:https://doi.org/10.1038/s42004-025-01501-6
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author Pasquale A. Morese
Ayaz Ahmad
Mathew P. Martin
Richard A. Noble
Sara Pintar
Lan Z. Wang
Shangze Xu
Andrew Lister
Richard A. Ward
Agnieszka K. Bronowska
Martin E. M. Noble
Hannah L. Stewart
Michael J. Waring
author_facet Pasquale A. Morese
Ayaz Ahmad
Mathew P. Martin
Richard A. Noble
Sara Pintar
Lan Z. Wang
Shangze Xu
Andrew Lister
Richard A. Ward
Agnieszka K. Bronowska
Martin E. M. Noble
Hannah L. Stewart
Michael J. Waring
author_sort Pasquale A. Morese
collection DOAJ
description Abstract The discovery of targeted covalent inhibitors is of increasing importance in drug discovery. Finding efficient covalent binders requires modulation of warhead reactivity and optimisation of warhead geometry and non-covalent interactions. Uncoupling the contributions that these factors make to potency is difficult and best practice for a testing cascade that is pragmatic and informative is yet to be fully established. We studied the structure-reactivity-activity relationships of a series of analogues of the EGFR inhibitor poziotinib with point changes in two substructural regions as well as variations in warhead reactivity and geometry. This showed that a simple probe displacement assay that is appropriately tuned in respect of timing and reagent concentrations can reveal structural effects on all three factors: non-covalent affinity, warhead reactivity and geometry. These effects include the detection of potency differences between an enantiomeric pair that differ greatly in their activity and their capacity to form a covalent bond. This difference is rationalised by X-ray crystallography and computational studies and the effect translates quantitatively into cellular mechanistic and phenotypic effects.
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spelling doaj-art-0f4e344ee95c402b81446b50b5a0be322025-08-20T03:08:24ZengNature PortfolioCommunications Chemistry2399-36692025-04-01811810.1038/s42004-025-01501-6Factors affecting irreversible inhibition of EGFR and influence of chirality on covalent bindingPasquale A. Morese0Ayaz Ahmad1Mathew P. Martin2Richard A. Noble3Sara Pintar4Lan Z. Wang5Shangze Xu6Andrew Lister7Richard A. Ward8Agnieszka K. Bronowska9Martin E. M. Noble10Hannah L. Stewart11Michael J. Waring12Cancer Research Horizons Newcastle Drug Discovery Unit, Chemistry, School of Natural and Environmental Sciences, Bedson Building, Newcastle UniversityChemistry, School of Natural and Environmental Sciences, Bedson Building, Newcastle UniversityCancer Research Horizons Newcastle Drug Discovery Unit, Translational and Clinical Research Institute, Paul O’Gorman Building, Newcastle UniversityCancer Research Horizons Newcastle Drug Discovery Unit, Translational and Clinical Research Institute, Paul O’Gorman Building, Newcastle UniversityCancer Research Horizons Newcastle Drug Discovery Unit, Translational and Clinical Research Institute, Paul O’Gorman Building, Newcastle UniversityCancer Research Horizons Newcastle Drug Discovery Unit, Translational and Clinical Research Institute, Paul O’Gorman Building, Newcastle UniversityChemistry, School of Natural and Environmental Sciences, Bedson Building, Newcastle UniversityOncology iMed, R&D, AstraZenecaOncology iMed, R&D, AstraZenecaChemistry, School of Natural and Environmental Sciences, Bedson Building, Newcastle UniversityCancer Research Horizons Newcastle Drug Discovery Unit, Translational and Clinical Research Institute, Paul O’Gorman Building, Newcastle UniversityCancer Research Horizons Newcastle Drug Discovery Unit, Chemistry, School of Natural and Environmental Sciences, Bedson Building, Newcastle UniversityCancer Research Horizons Newcastle Drug Discovery Unit, Chemistry, School of Natural and Environmental Sciences, Bedson Building, Newcastle UniversityAbstract The discovery of targeted covalent inhibitors is of increasing importance in drug discovery. Finding efficient covalent binders requires modulation of warhead reactivity and optimisation of warhead geometry and non-covalent interactions. Uncoupling the contributions that these factors make to potency is difficult and best practice for a testing cascade that is pragmatic and informative is yet to be fully established. We studied the structure-reactivity-activity relationships of a series of analogues of the EGFR inhibitor poziotinib with point changes in two substructural regions as well as variations in warhead reactivity and geometry. This showed that a simple probe displacement assay that is appropriately tuned in respect of timing and reagent concentrations can reveal structural effects on all three factors: non-covalent affinity, warhead reactivity and geometry. These effects include the detection of potency differences between an enantiomeric pair that differ greatly in their activity and their capacity to form a covalent bond. This difference is rationalised by X-ray crystallography and computational studies and the effect translates quantitatively into cellular mechanistic and phenotypic effects.https://doi.org/10.1038/s42004-025-01501-6
spellingShingle Pasquale A. Morese
Ayaz Ahmad
Mathew P. Martin
Richard A. Noble
Sara Pintar
Lan Z. Wang
Shangze Xu
Andrew Lister
Richard A. Ward
Agnieszka K. Bronowska
Martin E. M. Noble
Hannah L. Stewart
Michael J. Waring
Factors affecting irreversible inhibition of EGFR and influence of chirality on covalent binding
Communications Chemistry
title Factors affecting irreversible inhibition of EGFR and influence of chirality on covalent binding
title_full Factors affecting irreversible inhibition of EGFR and influence of chirality on covalent binding
title_fullStr Factors affecting irreversible inhibition of EGFR and influence of chirality on covalent binding
title_full_unstemmed Factors affecting irreversible inhibition of EGFR and influence of chirality on covalent binding
title_short Factors affecting irreversible inhibition of EGFR and influence of chirality on covalent binding
title_sort factors affecting irreversible inhibition of egfr and influence of chirality on covalent binding
url https://doi.org/10.1038/s42004-025-01501-6
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