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Specific targeting of neurotoxic side effects and pharmacological profile of the novel cancer stem cell drug salinomycin in mice

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Abstract

Salinomycin is a polyether antibiotic which effectively eliminates a variety of cancer stem cells and chemotherapy-resistant tumor cells in vitro and in vivo. One important caveat for its clinical application is the paucity of preclinical pharmacological and safety data. In the present study, we thus aimed to elucidate pharmacokinetic properties of salinomycin and to assess the side effect profile of chronic treatment with this compound in C57Bl/6 mice. In addition, we tested whether neurotoxic side effects can be prevented by interference with the intracellular calcium homeostasis. We observed that salinomycin has a narrow therapeutic index; however, a dose of 5 mg/kg body weight was well tolerated, and analysis of blood parameters as well as organ histology of liver, kidney, skeletal muscle, and heart showed no abnormalities after daily salinomycin injection for 4 weeks. Pharmacokinetic evaluation revealed low micromolar peak concentrations and an almost complete systemic elimination within 5 h after injection. In contrast to low systemic toxicity, typical signs of a sensory polyneuropathy with mechanical and cold allodynia, distinct gait alterations, decreased sensory nerve action potential amplitudes, and loss of myelinated fibers in the sciatic nerve were observed in salinomycin-treated animals. Inhibition of the mitochondrial Na+/Ca2+ exchanger partially prevented the development of salinomycin-induced neuropathy in vivo, an approach which did not reduce salinomycin’s antineoplastic efficacy in vitro. Taken together, this study establishes a framework of pharmacokinetic data for future preclinical trials and safety data for translational trials. Furthermore, we established a strategy to reduce salinomycin’s off-target neurotoxic effects.

Key message

  • Salinomycin has a narrow therapeutic index; a dose of 5 mg/kg is tolerated in mice.

  • Mice treated with salinomycin develop a painful sensory polyneuropathy.

  • An optimized protocol was established to measure salinomycin in serum samples.

  • Inhibition of Na+/Ca2+ exchangers prevents salinomycin-induced neuropathy.

  • Blocking mitochondrial Na+/Ca2+ exchangers does not impair antineoplastic efficacy.

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Acknowledgement

We would like to thank Catherine Aubel for the thoughtful text editing, Heather L. More for the Matlab routine used to perform semi-automatic nerve morphometry, and Petra Loge for the excellent technical assistance. We would also like to thank Thomas Kolrep for his commitment regarding the mass spectrometric experiments. The research leading to these results has received funding from the federal ministry of education and research via the grant center for stroke research Berlin (01 EO 0801), the Volkswagen foundation (Lichtenberg program to Matthias Endres), and the Deutsche Forschungsgemeinschaft DFG (NeuroCure, BioSupraMol). Wolfgang Boehmerle is participant in the Charité Clinical Scientist Program funded by the Charité Universitätsmedizin Berlin and the Berlin Institute of Health.

Disclosure

There are no known conflicts of interest with respect to the execution of the experiments or the preparation of the manuscript.

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Authors and Affiliations

  1. Klinik und Hochschulambulanz für Neurologie, Charité Universitätsmedizin Berlin, Chariteplatz 1, 10117, Berlin, Germany

    Wolfgang Boehmerle, Hanna Muenzfeld, Petra Huehnchen & Matthias Endres

  2. Cluster of Excellence NeuroCure, Charité Universitätsmedizin Berlin, Berlin, Germany

    Wolfgang Boehmerle, Petra Huehnchen & Matthias Endres

  3. Center for Stroke Research Berlin, Charité Universitätsmedizin Berlin, Berlin, Germany

    Matthias Endres

  4. German Centre for Neurodegenerative Diseases (DZNE), Chariteplatz 1, 10117, Berlin, Germany

    Matthias Endres

  5. Großgerätezentrum BioSupraMol, Department of Chemistry, Freie Universität Berlin, 14195, Berlin, Germany

    Andreas Springer

Authors
  1. Wolfgang Boehmerle
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  2. Hanna Muenzfeld
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  3. Andreas Springer
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  4. Petra Huehnchen
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  5. Matthias Endres
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Corresponding author

Correspondence to Wolfgang Boehmerle.

Additional information

Wolfgang Boehmerle and Hanna Muenzfeld contributed equally to the present manuscript.

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Supplemental Figure S1

Assessment of heat allodynia. Response latency in the hot plate test (50 °C) was comparable in all four experimental groups (GIF 27 kb)

High Resolution Image (TIFF 66 kb)

Supplemental Figure S2

MS/MS spectrum of deprotonated salinomycin. The fragment ions observed can be deduced from the fragmentation pattern suggested for sodiated salinomycin (GIF 47 kb)

High Resolution Image (TIFF 7,940 kb)

Supplemental Figure S3

SAL calibration standards with linear regression. Concentration values are in nanomole. r = 9.998847; r 2 = 0.997695 (GIF 9 kb)

High Resolution Image (TIFF 1,223 kb)

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Boehmerle, W., Muenzfeld, H., Springer, A. et al. Specific targeting of neurotoxic side effects and pharmacological profile of the novel cancer stem cell drug salinomycin in mice. J Mol Med 92, 889–900 (2014). https://doi.org/10.1007/s00109-014-1155-0

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  • DOI: https://doi.org/10.1007/s00109-014-1155-0

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