title: Experimental design on single-time-point high-throughput microsomal stability assay.
abstract: An experimental design for a single-time-point microsomal stability assay was evaluated as compared with multiple-time-point studies. Results obtained from single-time-point experiments are in excellent agreement with those from multiple time points. First-order reaction kinetics revealed rapid changes of predicted half-life from percent remaining of the parent compound at the inflection points, suggesting a maximum predictive limit for half-life. Selection of the incubation time in single-time-point assays is important to obtain balanced information for stable and unstable compounds. A short incubation time (e.g., 5 min) is most useful for differentiating between unstable compounds, which is beneficial to direct the synthetic efforts in projects with poor metabolic stability. A long incubation time (e.g., 30 min) is more applicable to a compound series with high metabolic stability. For screening purposes, a moderate incubation time (e.g., 15 min) is recommended to achieve good resolution and a sufficiently high maximum predictive limit for half-life. This study suggests that a single-time-point assay is sufficient for ranking compounds in early drug discovery. It increases throughput and reduces turnaround time and cost.
authors.affiliation: Wyeth Research, P.O. Box CN 8000, Princeton, New Jersey 08543-8000, USA. dil@wyeth.com
authors.forename: Li
authors.initials: L
authors.lastname: Di
authors.forename: Edward H
authors.initials: EH
authors.lastname: Kerns
authors.forename: Ning
authors.initials: N
authors.lastname: Gao
authors.forename: Susan Q
authors.initials: SQ
authors.lastname: Li
authors.forename: Youping
authors.initials: Y
authors.lastname: Huang
authors.forename: Jim L
authors.initials: JL
authors.lastname: Bourassa
authors.forename: Donna M
authors.initials: DM
authors.lastname: Huryn
mesh_terms: D000818:Animals; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism*; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012107:Research Design / Q000706:statistics & numerical data*
chemical_list: D004364:Pharmaceutical Preparations