Experimental design on single-time-point high-throughput microsomal stability assay
✍ Scribed by Li Di; Edward H. Kerns; Ning Gao; Susan Q. Li; Youping Huang; Jim L. Bourassa; Donna M. Huryn
- Publisher
- John Wiley and Sons
- Year
- 2004
- Tongue
- English
- Weight
- 105 KB
- Volume
- 93
- Category
- Article
- ISSN
- 0022-3549
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✦ Synopsis
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-timepoint 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 singletime-point assay is sufficient for ranking compounds in early drug discovery. It increases throughput and reduces turnaround time and cost.