MS Binding Assays - A New Concept for Drug Screening

Klaus Wanner, Full Professor, Ludwig- Maximilians-University Munich

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Launch presentation

About the speaker

Klaus T. Wanner studied chemistry and pharmacy at the TU and the Ludwig-Maximilians University in Munich. After obtaining his PhD he worked as a post-doc at Colorado State University, USA. Upon returning to Munich he completed his professorial thesis in 1988. In 1990 he accepted a position as professor at the Freie Universität Berlin, and returned to Ludwig-Maximilians University in 1994. , Prof. Wanner's research focuses on asymmetric synthesis as well as the development of inhibitors of the GABA transport proteins. Another important area of research are MS binding assays developed by the research group to replace radioligand binding assays.

Abstract

Bioluminescence is well established as a superior technology for predictive high throughput assays that are simple, sensitive, and cost effective and performed with low incidences of test compound interference. The availability of bioluminescent cytochrome P450 (CYP) assays overcomes limitations of conventional CYP assays and adds the bioluminescent advantages to the tool box for ADMET studies where CYP inhibition and induction assays are critical. Probe CYP substrates are used that are inactive luciferin derivatives. CYPs convert the substrates to active luciferin that is detected in a second reaction with a firefly luciferase that generates light in proportion to the amount of CYP enzyme activity. The predictive value of this approach was considered by comparisons to conventional methods. IC50s measured by the luminogenic method were found to correlate well in terms of rank order and absolute potency to those measured by fluorescent, LC/MS, LC/UV and radiometric methods. Likewise, homogenous bioluminescent cell based assays detected fold increases in CYP enzyme activities that correlate well to measurements made using conventional assays. These close correlations establish the value of in vitro bioluminescent CYP assays for predicting in vivo effects while harnessing the considerable advantages of a bioluminescent approach.

Launch presentation