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01-02-2015 | Original Article

Cellular uptake kinetics of bortezomib in relation to efficacy in myeloma cells and the influence of drug transporters

Authors: Jannick Clemens, Anja Seckinger, Dirk Hose, Dirk Theile, Magdalena Longo, Walter Emil Haefeli, Jürgen Burhenne, Johanna Weiss

Published in: Cancer Chemotherapy and Pharmacology | Issue 2/2015

Abstract

Purpose

Despite overall successful application to multiple myeloma patients, clinical efficacy of the proteasome inhibitor bortezomib is typically challenged by primary and secondary resistance of unknown origin. So far, the potential impact of intracellular concentrations on drug efficacy of bortezomib and the influence of drug transporters are unknown.

Methods

We determined cellular bortezomib kinetics in nine myeloma cell lines using ultrahigh-performance liquid chromatography coupled to tandem mass spectrometry. The potential influence of drug transporters on the uptake kinetics observed in these cell lines was investigated by testing substrate characteristics of bortezomib for several transporters in over-expressing model cells. Additionally, transporter mRNA expression was quantified in myeloma cell lines by real-time polymerase chain reaction (RT-PCR).

Results

All myeloma cells revealed an extensive intracellular bortezomib accumulation (47.5–183 ng/ml) exceeding extracellular concentrations (0.04–0.17 ng/ml) by more than factor 1,000. Only organic anion-transporting polypeptide 1B1 facilitated the uptake in over-expressing cells, however, to a negligible extent (factor 1.36). Bortezomib efflux via P-glycoprotein was confirmed by demonstrating reduced sensitivity (IC₅₀ 11.6 vs. 2.8 ng/ml) and intracellular concentrations (−56.1 %) in over-expressing cells compared to controls. RT-PCR revealed a varying but overall weak transporter expression in the studied myeloma cells without any correlation to intracellular concentrations. Although principally valid as demonstrated in the P-glycoprotein over-expressing cell model, there was no significant correlation between intracellular concentrations and bortezomib efficacy in myeloma cell lines.

Conclusion

Differences in intracellular concentrations in myeloma cell lines neither result from variable transporter expression nor represent the main factor determining bortezomib efficacy in vitro.

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Title: Cellular uptake kinetics of bortezomib in relation to efficacy in myeloma cells and the influence of drug transporters
Authors: Jannick Clemens
Anja Seckinger
Dirk Hose
Dirk Theile
Magdalena Longo
Walter Emil Haefeli
Jürgen Burhenne
Johanna Weiss
Publication date: 01-02-2015
Publisher: Springer Berlin Heidelberg
Published in: Cancer Chemotherapy and Pharmacology / Issue 2/2015
Print ISSN: 0344-5704
Electronic ISSN: 1432-0843
DOI: https://doi.org/10.1007/s00280-014-2643-1

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