Pitfalls and Considerations in Determining the Potency and Mutant Selectivity of Covalent Epidermal Growth Factor Receptor Inhibitors.

Autor: Hoyt KW; Department of Chemistry, University at Buffalo, The State University of New York, Buffalo, New York 14260, United States., Urul DA; AssayQuant Technologies, Inc., Marlboro, Massachusetts 01752, United States., Ogboo BC; Department of Chemistry, University at Buffalo, The State University of New York, Buffalo, New York 14260, United States., Wittlinger F; Department of Pharmaceutical and Medicinal Chemistry, Institute of Pharmaceutical Sciences, Eberhard Karls Universität Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany., Laufer SA; Department of Pharmaceutical and Medicinal Chemistry, Institute of Pharmaceutical Sciences, Eberhard Karls Universität Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany.; Cluster of Excellence iFIT (EXC 2180) 'Image-Guided and Functionally Instructed Tumor Therapies', Eberhard Karls Universität Tübingen, 72076 Tübingen, Germany.; Tübingen Center for Academic Drug Discovery & Development (TüCAD2), 72076 Tübingen, Germany., Schaefer EM; AssayQuant Technologies, Inc., Marlboro, Massachusetts 01752, United States., May EW; AssayQuant Technologies, Inc., Marlboro, Massachusetts 01752, United States., Heppner DE; Department of Chemistry, University at Buffalo, The State University of New York, Buffalo, New York 14260, United States.; Department of Pharmacology and Therapeutics, Roswell Park Comprehensive Cancer Center, Buffalo, New York 14203, United States.; Department of Structural Biology, The State University of New York, Buffalo, New York 14203, United States.
Jazyk: angličtina
Zdroj: Journal of medicinal chemistry [J Med Chem] 2024 Jan 11; Vol. 67 (1), pp. 2-16. Date of Electronic Publication: 2023 Dec 22.
DOI: 10.1021/acs.jmedchem.3c01502
Abstrakt: Enzyme inhibitors that form covalent bonds with their targets are being increasingly pursued in drug development. Assessing their biochemical activity relies on time-dependent assays, which are distinct and more complex compared with methods commonly employed for reversible-binding inhibitors. To provide general guidance to the covalent inhibitor development community, we explored methods and reported kinetic values and experimental factors in determining the biochemical activity of various covalent epidermal growth factor receptor (EGFR) inhibitors. We showcase how liquid handling and assay reagents impact kinetic parameters and potency interpretations, which are critical for structure-kinetic relationships and covalent drug design. Additionally, we include benchmark kinetic values with reference inhibitors, which are imperative, as covalent EGFR inhibitor kinetic values are infrequently consistent in the literature. This overview seeks to inform best practices for developing new covalent inhibitors and highlight appropriate steps to address gaps in knowledge presently limiting assay reliability and reproducibility.
Databáze: MEDLINE
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