MDCKII-BCRP

About the Assay

The MDCKII–BCRP (Madin–Darby canine kidney II) is a canine epithelial cell line stably transfected with human ATP binding cassette transporter protein BCRP (ABCG2). MDCKII-BCRP cells form tight, polarized monolayers suitable for drug transport assays and enable exploration of specific interactions with human BCRP. It has been reported that transfection of wild type MDCKII cells with human transporters results in a knock down of endogenous (canine) transporters including those responsible for efflux.

Drug transport is measured bi-directionally, apical-to-basolateral (AtoB) and basolateral-to-apical (BtoA), which allows for the rate of permeability to be determined and for the identification of BCRP mediated efflux substrates.

Given lipid bilayers are universally conserved across all species, MDCKII-BCRP cells represent a useful and human relevant system for bidirectional transport studies which enable the identification of passive and active transport processes and can help predict the oral absorption and CNS penetrance of new chemical entities.

Protocol Summary

Sygnature discovery use pre‑plated MDCKII‑BCRP cells (MedTech Barcelona) to assess compound permeability and efflux potential in hanging insert format.

Compound Selection

A diverse set of compounds was selected to span a range of intrinsic permeabilities and to include both efflux and non‑efflux substrates.

Validation Results

Figure 1  Apparent permeability coefficient (Papp) results for test compounds evaluated in Sygnature Discovery’s MDCKII BCRP assay with HBSS pH 7.4/7.4 buffer. Each bar represents the mean +/- SD Papp from individual experiments, A-B (top) and B-A (bottom), of validation compounds in order from lowest to highest.

Figure 2. Efflux ratio calculated for test compounds evaluated in Sygnature Discovery’s MDCKII BCRP assay with HBSS pH 7.4/7.4 buffer. The dotted line indicates the threshold of ER = 2. Results are presented as mean +/- SD

Figure 3. Efflux observed for test compounds evaluated in Sygnature Discovery’s MDCKII BCRP assay with HBSS pH 7.4/7.4 buffer. The dotted line indicates the threshold of ER = 2. Results are presented as mean +/- SD

References

Kuteykin-Teplyakov, K., Luna-Tortós, C., Ambroziak, K. and Löscher, W. (2010), Differences in the expression of endogenous efflux transporters in MDR1-transfected versus wildtype cell lines affect P-glycoprotein mediated drug transport. British Journal of Pharmacology, 160: 1453-1463.

The International Transporter Consortium, (2010), Nat Rev 9; 215-236.

Elsby, R., Coghlan, H., Edgerton, J., Hodgson, D., Outteridge, S. & Atkinson, H. (2023) Mechanistic in vitro studies indicate that the clinical drug–drug interactions between protease inhibitors and rosuvastatin are driven by inhibition of intestinal BCRP and hepatic OATP1B1 with minimal contribution from OATP1B3, NTCP and OAT3. Pharmacology Research & Perspectives, 11(2), e01060.

Li, J., Wang, Y., Zhang, W., Huang, Y., Hein, K. & Hidalgo, I.J. (2012) The role of a basolateral transporter in rosuvastatin transport and its interplay with apical breast cancer resistance protein in polarized cell monolayer systems. Drug Metabolism and Disposition, 40(11), 2102–2108