Poller, Birk, Werner, Sophie, Domange, Norbert, Bednarczyk, Dallas, Mettler, Lina, Stein, Richard, Loretan, Jacqueline, Wartmann, Markus, Faller, Bernard and Huth, Felix (2022) Time matters – In vitro cellular disposition kinetics help identifying disconnects in cell-based potency assays. Xenobiotica, 52 (8). pp. 878-889. ISSN https://doi.org/10.1080/00498254.2022.2130837

Abstract

1. Decreasing potency is commonly observed in early drug discovery when moving from biochemical to more complex cellular systems. Among other factors low permeability is often considered to cause such potency disconnects.
2. We developed a novel cellular disposition assays in MDCK cells to determine initial uptake rate (PSinf), cell-to-medium ratios under steady-state (Kp) and the time to reach equilibrium (TTSS90) from a single experiment in a high throughput assay.
3. The assay was validated using 40 marketed drugs, showing a wide distribution of PSinf and Kp data. The parameters generally correlated with transwell permeability and lipophilicity (logD), while PSinf data revealed better resolution in the high and low permeability ranges. A linear correlation between the Kp/PSinf ratio and TTSS90 was found, demonstrating the dependency of TTSS90 on the rate and extent of a compound to accumulate in cells.
4. Cellular disposition parameters could explain potency (IC50) disconnects for eight BTK degrader compounds in a cellular potency assay. In contrast to transwell permeability, PSinf data enabled differentiation of the compounds with and without IC50 disconnects based on their time to reach equilibrium. Overall, the novel assay offers the possibililty to address potency disconnects during an early drug discovery phase.

Item Type:	Article
Date Deposited:	07 Mar 2023 00:45
Last Modified:	07 Mar 2023 00:45
URI:	https://oak.novartis.com/id/eprint/47952