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Rapid Determination of the Specificity Constant of Irreversible Inhibitors ( k inact / K I ) by Means of an Endpoint Competition Assay
Author(s) -
Miyahisa Ikuo,
Sameshima Tomoya,
Hixon Mark S.
Publication year - 2015
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201505800
Subject(s) - chemistry , ic50 , potency , biophysics , biochemistry , in vitro , biology
Owing to their covalent target occupancy, irreversible inhibitors require low exposures and offer long duration, and their use thus represents a powerful strategy for achieving pharmacological efficacy. Importantly, the potency metric of irreversible inhibitors is k inact / K I not IC 50 . A simple approach to measuring k inact / K I was developed that makes use of an irreversible probe for competitive assays run to completion against test compounds. In this system, the k inact / K I value of the test compound is equal to ( k inact / K I ) probe ×[probe]/IC 50 . The advantages of this method include simplicity, high throughput, and application to all target classes, and it only requires an in‐depth kinetic evaluation of the probe.