Development of hydrolytic stability screening methods for early drug discovery with high differentiation and...

In early drug discovery, hydrolytic chemical stability is routinely assessed to ensure future developability of quality compounds and stability in in vitro test environments. When conducting high-throughput hydrolytic stability analyses as part of the compound risk assessment, aggressive conditions are typically applied to allow for faster screening. However, it can be challenging to extrapolate the real stability risk and to rank compounds due to over-estimating risk based on aggressive conditions and the narrow discriminative window. In this study, critical assay parameters including temperature, concentration, and detection technique were systematically assessed using selected model compounds, and the impact and interplay of these parameters on predictive power and prediction quality were evaluated. Improved data quality was achieved using high sample concentration and reduced temperature, combined with ultraviolet (UV) detection, while mass spectrometry (MS) detection was found to be a useful complementary detection technique. Therefore, a highly discriminative stability protocol with optimized assay parameters and experimental data quality is proposed. The optimized assay can provide early guidance on the potential stability risk of a drug molecule as well as enable more confident decision-making in compound design, selection, and development.