Demonstrating suitability of the Caco-2 cell model for BCS-based biowaiver according to the recent FDA and ICH harmonised guidelines

Abstract

Objective According to the regulatory guidelines, one of the critical steps in using in‐vitro permeability methods for permeability classification is to demonstrate the suitability of the method. Here, suitability of the permeability method by using a monolayer of cultured epithelial cells was verified with different criteria. Methods Imaging with a transmission electron microscope was used for characterisation of the cells. Monolayer integrity was confirmed by transepithelial electrical resistance measurements and permeability of zero permeability marker compounds. Real‐time polymerase chain reaction was employed to evaluate expression levels of 84 known transporters. Samples for bidirectional permeability determination were quantified by ultra‐performance liquid chromatography. Key findings The Caco‐2 cells grow in an intact monolayer and morphologically resemble enterocytes. Genes of 84 known transporters were expressed at different levels; furthermore, expression was time depended. Functional expression of efflux transporter P‐glycoprotein was confirmed. We established a correlation between permeability coefficients of 21 tested drug substances ranging from low, moderate and high absorption with human fraction absorbed literature data (R2 = 0.84). Conclusions Assay standardisation assures the consistency of experimental data. Only such fully characterised model has the ability to accurately predict drug's intestinal permeability at the early stage of research or for the BCS‐based biowaiver application.

Biopharmaceutical classification of desloratadine – not all drugs are classified the easy way

“…This means that potential active transport in the intestine does not have any significant role in limiting desloratadine absorption. This is in line with our permeability data, since the efflux ratio is well below 2 even when measured by an in vitro model, which is known to over-express P-gp efflux transporter (21).…”

Single-cell transcriptomics elucidates in vitro reprogramming of human intestinal epithelium cultured in a physiodynamic gut-on-a-chip

“…The Caco-2 human colon adenocarcinoma line has been extensively used in biomedical studies and considered as a gold standard for estimating intestinal permeability of drugs in pharmaceutical validation over the past decades [5][6][7] . The Caco-2 epithelium demonstrates enterocyte-like phenotype with transcriptomic signatures of normal small intestinal epithelium when cultured for 3 weeks in a static condition despite its cancer origin [8][9][10] .…”

Comparative analysis of biopharmaceutic classification system (BCS) based biowaiver protocols to validate equivalence of a multisource product

“…Certain excipients reduce the GI transit period, therefore, GI transit period serves as a critical factor for bioequivalence proposing more strict criteria for dissolution to make sure complete dissolution in the stomach (Koch et al, 1993). Apart from in-vivo pharmacokinetic studies, presently only Caco-2 permeability studies are endorsed in the ICH harmonised guideline (ICH, 2018;Jarc et al, 2019). A study conducted to determine consequence of Caco-2 permeability for some formulation excipients showed that permeability across Caco-2 monolayers did not aggravate by such excipients (Rege et al, 2001).…”