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Characterization of Substrates and Inhibitors for the In Vitro Assessment of Bcrp Mediated Drug–Drug Interactions

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Abstract

Purpose

In vitro assessment of drug candidates' affinity for multi-drug resistance proteins is of crucial importance for the prediction of in vivo pharmacokinetics and drug–drug interactions. To have well described experimental tools at hand, the objective of the study was to characterize substrates and inhibitors of Breast Cancer Resistance Protein (BCRP) and P-glycoprotein (P-gp).

Methods

Madin–Darbin canine kidney cells overexpressing mouse Bcrp (MDCKII-Bcrp) were incubated with various Bcrp substrates, or a mixture of substrate and inhibitor to either the apical (A) or basolateral (B) compartment of insert filter plates. Substrate concentrations in both compartments at time points t = 0 h and t = 2 h were determined by LC–MS/MS, and respective permeation coefficients (P app) and efflux ratios were calculated.

Results

The Bcrp inhibitor Ko143 blocked topotecan and ABZSO transport in a concentration-dependent manner. P-gp inhibitors ivermectin, LY335979, PSC833, and the P-gp/Bcrp inhibitor ritonavir did not influence Bcrp mediated topotecan transport, however, blocked ABZSO transport. Additionally, neither was ABZSO transport influenced by topotecan, nor topotecan transport by ABZSO.

Conclusions

Data suggest different modes of substrate and inhibitor binding to Bcrp. In order to not overlook potential drug–drug interactions when testing drug candidates for inhibitory potential towards Bcrp, distinct Bcrp probe substrates should be used.

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Abbreviations

A:

apical

ABCB1:

ATP binding cassette transporter B1

ABCG2:

ATP binding cassette transporter G2

ABCP:

human placenta-specific ATP-binding cassette gene

ABZSO:

albendazole sulphoxide

B:

basolateral

BCRP:

breast cancer resistance protein

DMEM:

Dulbecco's modified Eagle’s medium

DMSO:

dimethylsulfoxide

FCS:

fetal calf serum

HBSS:

Hanks’ balanced salt solution

HEPES:

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid)

LLC-PK1:

porcine kidney tubular epithelial cell line

MDCKII:

Madin Darby canine kidney cell line II

MXR:

mitoxantrone resistance gene

P app :

apparent permeability coefficient

P-gp:

P-glycoprotein

TEER:

transepithelial electrical resistance

wt:

wild-type

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Acknowledgements

We thank Mark Twele for technical assistance during LC–MS/MS analytics.

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Authors and Affiliations

  1. Department of Metabolism and Pharmacokinetics, Bayer HealthCare AG, Aprather Weg, 42096, Wuppertal-Aprath, Germany

    Uwe Muenster, Birgit Grieshop, Karsten Ickenroth & Mark Jean Gnoth

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  1. Uwe Muenster
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  2. Birgit Grieshop
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Correspondence to Mark Jean Gnoth.

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Muenster, U., Grieshop, B., Ickenroth, K. et al. Characterization of Substrates and Inhibitors for the In Vitro Assessment of Bcrp Mediated Drug–Drug Interactions. Pharm Res 25, 2320–2326 (2008). https://doi.org/10.1007/s11095-008-9632-1

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  • DOI: https://doi.org/10.1007/s11095-008-9632-1

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