Comparative study of absorption prediction using human intestinal organoid-derived cells in static and flow...

Reliable prediction of human oral drug absorption is critical in early-stage drug development. However, physiologically relevant intestine cell models capturing multiple drug disposition mechanisms including transport and metabolism are limited. This study assessed the potential of human intestinal organoid-derived epithelial monolayers to characterize drug permeability and absorption in 2 culture formats-a traditional static Transwell system and a dynamic fluidic gut-on-chip system. Initially, dynamic gene and protein expression profiles were characterized over 5 weeks of static culture, followed by assessments of functional permeability using control compounds. Subsequently, the effects of mechanical and chemical stimuli were explored under dynamic flow conditions. Results indicated that although the integrity of cell monolayers in static cultures remains intact for up to 5 weeks in differentiation medium, cell differentiation stages, and the expression of specific genes and proteins related to drug metabolism exhibit time-dependent variations. Fluidic cultures showed enhanced expression of drug disposition-related genes and proteins, along with relatively higher apparent permeability. Both formats demonstrated strong correlations between the measured apparent permeability and human effective permeability, and moreover, predicted fraction of drug escaping intestinal extraction well. These findings support the application of human intestinal organoid-derived intestinal epithelial models as a reliable tool to enable human absorption predictions with reduced reliance on animal models. SIGNIFICANCE STATEMENT: This study offers significant advancements in the field of oral drug absorption prediction by leveraging human intestinal organoid-derived epithelial monolayers in both static and dynamic culture systems. By demonstrating improved physiological relevance and strong correlations with human absorption parameters, the research provides a robust alternative to traditional animal and cell models. These findings support the potential application of human intestinal organoid-derived models to reduce reliance on animal testing and improve translational accuracy in early-stage drug development.