Fluorescently Labeled Analogues of Dofetilide as High-Affinity Fluorescence Polarization Ligands for the Human Ether-a-go-go-Related Gene (hERG) Channel
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- 作者：David H. Singleton ; Helen Boyd ; Jill V. Steidl-Nichols ; Matt Deacon ; Marcel J. de Groot ; David Price ; David O. Nettleton ; Nora K. Wallace ; Matthew D. Troutman ; Christine Williams ; James G. Boyd
- 刊名：Journal of Medicinal Chemistry
- 出版年：2007
- 出版时间：June 28, 2007
- 年：2007
- 卷：50
- 期：13
- 页码：2931 - 2941
- 全文大小：318K
- 年卷期：v.50,no.13(June 28, 2007)
- ISSN：1520-4804
文摘
Novel fluorescent derivatives of dofetilide (1) have been synthesized. Analogues that feature a fluorescentprobe attached through an aliphatic spacer to the central tertiary nitrogen of 1 have high affinity for thehERG channel, and affinity is dependent on both linker length and pendent dye. These variables have beenoptimized to generate Cy3B derivative 10e, which has hERG channel affinity equivalent to that of dofetilide.When bound to cell membranes expressing the hERG channel, 10e shows a robust increase in fluorescencepolarization (FP) signal. In a FP binding assay using 10e as tracer ligand, Ki values for several knownhERG channel blockers were measured and excellent agreement with the literature Ki values was observedover an affinity range of 2 nM to 3 
M. 10e blocks hERG channel current in electrophysiological patchclamp experiments, and computational docking experiments predict that the dofetilide core of 10e bindshERG channel in a conformation similar to that previously predicted for 1. These analogues enable high-throughput hERG channel binding assays that are rapid, economical, and predictive of test compounds'potential for prolonged QT liabilities.
M. 10e blocks hERG channel current in electrophysiological patchclamp experiments, and computational docking experiments predict that the dofetilide core of 10e bindshERG channel in a conformation similar to that previously predicted for 1. These analogues enable high-throughput hERG channel binding assays that are rapid, economical, and predictive of test compounds'potential for prolonged QT liabilities.