title: Discovery and development of an aurora kinase inhibitor clinical candidate using an image-based assay for measuring proliferation, apoptosis, and DNA content.
abstract: Measurement of antiproliferative capacity of a compound is central to early oncology drug discovery, with information about the precise mechanism of compound action typically being acquired during later downstream assays. Here we describe the development and validation of an in vitro image-based assay that simultaneously measures tumor cell count, late apoptotic morphology, and nuclear DNA content (termed the proliferation, apoptosis, and DNA content [PAD] assay) by using a DNA binding fluorescent dye. The PAD assay determines whether a compound's antiproliferative effect occurs via cell cycle arrest or induction of apoptosis, replacing downstream assays for 1/50(th) the cost. We used this assay to screen a kinase inhibitor-biased library and discovered an Aurora kinase inhibitor, and we also used it to drive structure-activity relationship to clinical candidate Investigational New Drug filing within 2 years. The simplicity of the PAD assay was critical to the rapid time frame within which this candidate was identified and progressed. This inhibitor is currently beginning Phase II clinical trials.
authors.affiliation: Rigel Pharmaceuticals, Inc., South San Francisco, California, USA. scatalano@drugdiscoveryimaging.com
authors.forename: Susan M
authors.initials: SM
authors.lastname: Catalano
authors.forename: John
authors.initials: J
authors.lastname: McLaughlin
authors.forename: Yasumichi
authors.initials: Y
authors.lastname: Hitoshi
authors.forename: Don
authors.initials: D
authors.lastname: Payan
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D064127:Aurora Kinases; D001973:Bromodeoxyuridine / Q000378:metabolism; D002455:Cell Division; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004247:DNA / Q000032:analysis*; D016195:G2 Phase; D006801:Humans; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors*
chemical_list: D047428:Protein Kinase Inhibitors; D004247:DNA; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases; D001973:Bromodeoxyuridine