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
✍ Scribed by Sabrina X. Zhao; Darcy Forman; Nora Wallace; Bill J. Smith; Daniel Meyer; Diana Kazolias; Feng Gao; John Soglia; Mark Cole; David Nettleton
- Publisher
- John Wiley and Sons
- Year
- 2005
- Tongue
- English
- Weight
- 175 KB
- Volume
- 94
- Category
- Article
- ISSN
- 0022-3549
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✦ Synopsis
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 timepoint method samples taken at t ¼ 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.