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Interaction of the mitotic kinesin Eg5 inhibitor monastrol with P-glycoprotein

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Abstract

Monastrol is the first characterised small molecule inhibitor of the motor protein Eg5 involved in bipolar mitotic spindle assembly. Eg5 localises to microtubules in mitosis, but not to interphase microtubules, suggesting that Eg5 inhibitors may be useful to specifically target proliferating tumour tissue, thereby avoiding dose-limiting neuropathy observed with other antimicrotubule agents like taxanes or vinca alkaloids. Because other antimicrotubule agents fail in multidrug resistance associated with P-glycoprotein (Pgp) over-expression, we investigated the interaction of monastrol with Pgp in vitro. By means of the calcein assay (with P388/dx cells and primary porcine brain capillary endothelial cells) and confocal laser-scanning microscopy (with L-MDR1 cells) we demonstrated that monastrol is a weak inhibitor of Pgp in vitro, with f2 values being about two orders of magnitude greater than those of the well-known inhibitors verapamil and quinidine. Monastrol also induces Pgp in vitro as measured by mRNA expression in LS180 cells after incubation with monastrol. However, its effect is weak compared to rifampicin. Whilst it reveals weak inhibitory and inductive characteristics, monastrol appears to be not transported by Pgp, as indicated by the lack of difference in the antiproliferative effect of this compound in cell lines with and without over-expression of Pgp. The observed interaction profile of monastrol with Pgp is promising for the development of other more potent Eg5 inhibitors.

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Abbreviations

ANOVA:

analysis of variance

β2 mg:

β2-microglobulin

BCRP:

breast cancer resistance protein

calcein-AM:

calcein-acetoxymethylester

DMSO:

dimethyl sulfoxide

FCS:

foetal calf serum

f2:

concentration needed to double baseline fluorescence

G6PDH:

glucose-6-phosphate dehydrogenase

HBSS:

Hank’s balanced salt solution

HHBSS:

with HEPES supplemented HBSS

HEPES:

N-2-hydroxyethylpiperazine-N’-2-ethansulfonicacid

LY335979:

zosuquidar

MDR:

multidrug resistance

MRP:

multidrug resistance associated protein

MTT:

methylthiazolyldiphenyl-tetrazolium bromide

PBS:

phosphate buffered saline

Pgp:

P-glycoprotein

PXR:

pregnane x receptor

pBCECs:

porcine brain capillary endothelial cells

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Acknowledgements

We would like to thank Dr. Dario Ballinari for providing the cell lines P388 and P388/dx, Dr. Alfred Schinkel for providing the cell line L-MDR1, and Eli Lilly Company for providing LY335979. Moreover, we would like to thank Dr. Frank Zenke for his scientific review and for providing the test compounds monastrol and paclitaxel, Dr. Michael M. Hoffmann for sequencing of PCR products, as well as Stephanie Rosenzweig, Heiner Sähr, and Alexandra Sauer for their excellent technical assistance.

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Authors and Affiliations

  1. Department of Internal Medicine VI, Clinical Pharmacology and Pharmacoepidemiology, University of Heidelberg, Im Neuenheimer Feld 410, D-69120, Heidelberg, Germany

    Tanja Peters, Heike Lindenmaier, Walter E. Haefeli & Johanna Weiss

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  1. Tanja Peters
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  2. Heike Lindenmaier
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  3. Walter E. Haefeli
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  4. Johanna Weiss
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Correspondence to Johanna Weiss.

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Peters, T., Lindenmaier, H., Haefeli, W.E. et al. Interaction of the mitotic kinesin Eg5 inhibitor monastrol with P-glycoprotein. Naunyn Schmied Arch Pharmacol 372, 291–299 (2006). https://doi.org/10.1007/s00210-005-0022-5

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  • DOI: https://doi.org/10.1007/s00210-005-0022-5

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