Evaluate Compound Toxicity Early in the Drug Discovery Process Using High Content Analysis in Cell-Based Assays

Test compounds may have undesired side affects in addition to the desired modulation of a specific signaling pathway. Here we describe the use of high content cell-based assays to monitor such effects. In addition to the primary activity readout, many image analysis algorithms used for high content analysis report secondary readouts describing the cellular morphology, including number of cells, cell size and rounding, and nuclear morphology. Cell loss or changes in cell morphology as a consequence of compound treatment indicate acute compound cytotoxicity, and significant changes in the nuclear morphology are a sign of apoptosis and/or DNA damage. It is quite common that compounds which induce cytotoxic effects emerge as hits in small molecule screens and profiling studies. Such compounds can be discarded as false positives if secondary high content parameters are fully exploited. In this study, we show that this strategy may be applied to the Redistribution technology.