title: Simple strategies for reducing sample loads in in vitro metabolic stability high-throughput screening experiments: a comparison between traditional, two-time-point and pooled sample analyses.
abstract: Higher-throughput ADME programs in early drug discovery are becoming common throughout the pharmaceutical industry as companies strive to reduce their compound attrition in later-stage development. Many of the ADME assays developed into higher-throughput formats rely on LC/MS analyses. Since the biological aspects of the assay are amenable to parallel processes using dense plate formats, the number of samples generated from these assays produce a large analysis load for serial LC/MS. Presented in this report are two novel strategies, including a sample pooling method and a two time-point method, that could be used in drug discovery to reduce the number of samples generated during multiple time-point in-vitro ADME assays. One hundred and sixty-three compounds were subjected to human microsomal incubations with full time-point method samples taken at t = 0, 5, 15, 30, and 45 min. The ER data correlation (R(2)) between the full time-point method and the pooling method and two time-point methods were 0.98 and 0.97, respectively. Both methods have the potential to: 1. produce data of similar quality to traditional high throughput ADME assays, 2. be easily implemented, 3. shorten analytical run times, and 4. be reproducible and robust.
authors.affiliation: Candidate Enhancement Group, Exploratory Medicinal Sciences, Pfizer Inc., Eastern Point Road MS-8118W-102, Groton, Connecticut 06340, USA. sabrina_x_zhao@groton.pfizer.com <sabrina_x_zhao@groton.pfizer.com>
authors.forename: Sabrina X
authors.initials: SX
authors.lastname: Zhao
authors.forename: Darcy
authors.initials: D
authors.lastname: Forman
authors.forename: Nora
authors.initials: N
authors.lastname: Wallace
authors.forename: Bill J
authors.initials: BJ
authors.lastname: Smith
authors.forename: Daniel
authors.initials: D
authors.lastname: Meyer
authors.forename: Diana
authors.initials: D
authors.lastname: Kazolias
authors.forename: Feng
authors.initials: F
authors.lastname: Gao
authors.forename: John
authors.initials: J
authors.lastname: Soglia
authors.forename: Mark
authors.initials: M
authors.lastname: Cole
authors.forename: David
authors.initials: D
authors.lastname: Nettleton
mesh_terms: D000465:Algorithms; D002851:Chromatography, High Pressure Liquid; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D003627:Data Interpretation, Statistical; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006207:Half-Life; D006801:Humans; D007700:Kinetics; D013058:Mass Spectrometry; D008862:Microsomes, Liver / Q000201:enzymology; D004364:Pharmaceutical Preparations / Q000378:metabolism*
chemical_list: D004364:Pharmaceutical Preparations; D003577:Cytochrome P-450 Enzyme System