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Updated: Aug 26, 2025

Mass Spectrometry and Luminogenic-based Approaches to Characterize Phase I Metabolic Competency of In Vitro Cell Cultures
Published on: March 28, 2017
Time matters - in vitro cellular disposition kinetics help rationalizing cellular potency disconnects.
Birk Poller1, Sophie Werner1, Norbert Domange1
1Pharmacokinetic Sciences, Novartis Institutes for Biomedical Research, Basel, Switzerland.
A new assay measures cellular drug uptake, revealing how drug potency changes between biochemical and cellular tests. This helps identify drugs with potency disconnects early in discovery.
Area of Science:
- Pharmacology
- Drug Discovery
- Cell Biology
Background:
- Drug potency often decreases from biochemical to cellular assays.
- Low permeability is a key factor contributing to these potency disconnects.
- Accurate prediction of cellular drug behavior is crucial for early drug discovery.
Purpose of the Study:
- To develop a novel, high-throughput cellular disposition assay.
- To measure passive uptake clearance (PSinf), steady-state cell-to-medium ratio (Kp), and time to reach 90% steady-state (TTSS90).
- To address potency disconnects observed in early drug discovery.
Main Methods:
- Developed a novel cellular disposition assay in MDCK cells.
- Validated the assay using 40 marketed drugs.
- Determined PSinf, Kp, and TTSS90 in a single high-throughput experiment.
Main Results:
- The assay successfully measured PSinf, Kp, and TTSS90 across a range of marketed drugs.
- PSinf data provided better resolution for high and low permeability compounds than traditional methods.
- Cellular disposition parameters explained potency disconnects for Bruton's tyrosine kinase inhibitors.
Conclusions:
- The novel assay effectively characterizes cellular drug disposition.
- PSinf and TTSS90 data can identify compounds with potency disconnects.
- This assay aids in addressing potency disconnects in early drug discovery.
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