A fast screening model for drug permeability assessment based on native small intestinal extracellular matrix

Abstract

The Caco-2 cell monolayer model is widely utilized to predict drug permeability across human intestinal epithelial cells. However, at least 21 days is required for the formation and maturation of a well-tight Caco-2 cell monolayer, thereby restricting the throughput of the screening model during drug discovery. To address this challenge, a fast (7 days), and more physiologically relevant screening model integrating both the Caco-2 cell model and a small intestinal submucosa (SIS) hydrogel was developed in this study. The 7 day model exhibited desirable phenotype and functional similarity to the conventional 21 day Caco-2 model with respect to paracellular resistance, alkaline phosphatase (ALP) activities, and the mRNA expression level of three transporters (PEPT1, OATP1A2, and P-gp) as well as their mediated influx or efflux. Besides, the increased gene expression of two excretive transporters (BCRP, MRP2) and their enhanced functionality were observed in the current fast model compared to the traditional 21 day model. More importantly, a strong correlation (r² = 0.9458) was obtained between the absorptive P_app values of 19 model compounds in the 7 day model and those in the conventional 21 day model. These results revealed the pivotal role of the native extracellular matrix (SIS) in facilitating the differentiation of Caco-2 cells, leading to the reconstruction of the accelerated 7 day model, which presents a promising tool for screening drug permeability in future drug discovery.