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Abstract
PurposeTo examine the utility of human plasma as an assay medium in Caco-2 permeability studies to overcome poor mass balance and inadequate sink conditions frequently encountered with lipophilic compounds.MethodsCaco-2 permeability was assessed for reference compounds with known transport mechanisms using either pH 7.4 buffer or human plasma as the assay medium in both the apical and basolateral chambers. When using plasma, Papp values were corrected for the unbound fraction in the donor chamber. The utility of the approach was assessed by measuring the permeability of selected antimalarial compounds using the two assay media.ResultsCaco-2 cell monolayer integrity and P-gp transporter function were unaffected by the presence of human plasma in the donor and acceptor chambers. For many of the reference compounds having good mass balance with buffer as the medium, higher Papp values were observed with plasma, likely due to improved acceptor sink conditions. The lipophilic antimalarial compounds exhibited low mass balance with buffer, however the use of plasma markedly improved mass balance allowing the determination of more reliable Papp values.ConclusionsThe results support the utility of human plasma as an alternate Caco-2 assay medium to improve mass balance and permeability measurements for lipophilic compounds.
Highlights
In vitro permeability methods based on cell-based systems (e.g. Caco-2 and MDCK), isolated tissues, or artificial membranes (PAMPA) are well-established for testing the likely absorption properties of new drug candidates [1]
The results support the utility of human plasma as an alternate Caco-2 assay medium to improve mass balance and permeability measurements for lipophilic compounds
Compound binding was assessed at a similar concentration to that used in the donor chamber for permeability assessment to take into account the potential for saturable binding at high concentration
Summary
In vitro permeability methods based on cell-based systems (e.g. Caco-2 and MDCK), isolated tissues, or artificial membranes (PAMPA) are well-established for testing the likely absorption properties of new drug candidates [1]. The Caco-2 model is recommended by the FDA and EMA as one of the cell-based screening strategies to inform the need for in vivo transporter-based drug-drug interaction studies [5,6]. Pharm Res (2018) 35: 210 parameters for in silico prediction of compound absorption via physiologically-based pharmacokinetic (PBPK) models, and the FDA acknowledges the use of Caco-2 and other cultured epithelial cell monolayer test systems to assess permeability in relation to the biopharmaceutics classification system (BCS) and associated bio-waiver applications [7,8]. Compound permeability is a key component of the biopharmaceutics drug disposition classification system (BDDCS) [9], the extended clearance classification system (ECCS) [10], and the developability classification system (DCS) [11], each of which can be used to tailor compound profiling procedures during drug discovery and development
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