Identification of a small molecule that induces mitotic arrest using a simplified high-content screening assay and data analysis method

Abstract

High-content screening has emerged as a newand powerful technique for identifying small-molecule modulators ofmammalian cell biology. The authors describe the development and execution of a high-content screen to identify smallmolecules that induce mitotic arrest in mammalian cancer cells. Many widely used chemotherapeutics, such as Taxol® and vinblastine, induce mitotic arrest, and the creation of new drugs that also induce mitotic arrest may have tremendous therapeutic value. In their screen, the authors employed a simple DNA stain (DAPI) and a sensitive nonparametric statistical test to identify compounds from an internal collection of ∼13,000 high-quality lead-like small molecules. Subsequent analysis of 1 active compound indicated that it induces mitotic arrest, assessed using a high-content phosphohistone H3 detection assay, and caused cell proliferation defects inmultiple cancer cell lines. The active compound, a quinazolinone originating from a natural product-like subset of the screened compounds, is active in cells at ∼500nM and appears to act by inhibiting the polymerization of tubulin. © 2006 Society for Biomolecular Sciences.