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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
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201505800
Subject(s) - metric (unit) , chemistry , combinatorial chemistry , computer science , engineering , operations management
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.