Determination of Affinity and Residence Time of Potent Drug-Target Complexes by Label-free Biosensing

Journal of Medicinal Chemistry
John G QuinnMelinda M Mulvihill

Abstract

Prolonged drug-target occupancy has become increasingly important in lead optimization, and biophysical assays that measure residence time are in high demand. Here we report a practical label-free assay methodology that provides kinetic and affinity measurements suitable for most target classes without long preincubations and over comparatively short sample contact times. The method, referred to as a "chaser" assay, has been applied to three sets of unrelated kinase/inhibitor panels in order to measure the residence times, where correlation with observed efficacy was suspected. A lower throughput chaser assay measured a residence time of 3.6 days ±3.4% (95% CI) and provided single digit pM sensitivity. A higher throughput chaser methodology enabled a maximum capacity of 108 compounds in duplicate/day with an upper residence time limit of 9 h given an assay dissociation time of 34 min.

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Citations

Oct 2, 2019·ACS Central Science·Kin Sing Stephen LeeBruce D Hammock
Jul 2, 2019·Journal of Chemical Information and Modeling·Liang-Yong XiaYong Liang

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