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Biological Chemistry

Editor-in-Chief: Brüne, Bernhard

Editorial Board: Buchner, Johannes / Lei, Ming / Ludwig, Stephan / Thomas, Douglas D. / Turk, Boris / Wittinghofer, Alfred


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Volume 396, Issue 6-7

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Subcellular distribution of FTY720 and FTY720-phosphate in immune cells – another aspect of Fingolimod action relevant for therapeutic application

Matthias Schröder / Olga Arlt
  • Pharmazentrum frankfurt/ZAFES, Institute of General Pharmacology and Toxicology, Goethe University Frankfurt, Building 75, Theodor-Stern-Kai 7, D-60590 Frankfurt/Main, Germany
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/ Helmut Schmidt
  • Pharmazentrum frankfurt/ZAFES, Institute of Clinical Pharmacology, Goethe University, D-60590 Frankfurt/Main, Germany
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/ Andrea Huwiler / Carlo Angioni
  • Pharmazentrum frankfurt/ZAFES, Institute of Clinical Pharmacology, Goethe University, D-60590 Frankfurt/Main, Germany
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/ Josef M. Pfeilschifter
  • Pharmazentrum frankfurt/ZAFES, Institute of General Pharmacology and Toxicology, Goethe University Frankfurt, Building 75, Theodor-Stern-Kai 7, D-60590 Frankfurt/Main, Germany
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/ Anja Schwiebs
  • Corresponding author
  • Pharmazentrum frankfurt/ZAFES, Institute of General Pharmacology and Toxicology, Goethe University Frankfurt, Building 75, Theodor-Stern-Kai 7, D-60590 Frankfurt/Main, Germany
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/ Heinfried H. Radeke
  • Pharmazentrum frankfurt/ZAFES, Institute of General Pharmacology and Toxicology, Goethe University Frankfurt, Building 75, Theodor-Stern-Kai 7, D-60590 Frankfurt/Main, Germany
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Published Online: 2015-01-14 | DOI: https://doi.org/10.1515/hsz-2014-0287

Abstract

FTY720 (Fingolimod; Gilenya®) is an immune-modulatory prodrug which, after intracellular phosphorylation by sphingosine kinase 2 (SphK2) and export, mimics effects of the endogenous lipid mediator sphingosine-1-phosphate. Fingolimod has been introduced to treat relapsing-remitting multiple sclerosis. However, little has been published about the immune cell membrane penetration and subcellular distribution of FTY720 and FTY720-P. Thus, we applied a newly established LC-MS/MS method to analyze the subcellular distribution of FTY720 and FTY720-P in subcellular compartments of spleen cells of wild type, SphK1- and SphK2-deficient mice. These studies demonstrated that, when normalized to the original cell volume and calculated on molar basis, FTY720 and FTY720-P dramatically accumulated several hundredfold within immune cells reaching micromolar concentrations. The amount and distribution of FTY720 was differentially affected by SphK1- and SphK2-deficiency. On the background of recently described relevant intracellular FTY720 effects in the nanomolar range and the prolonged application in multiple sclerosis, this data showing a substantial intracellular accumulation of FTY720, has to be considered for benefit/risk ratio estimates.

Keywords: lymphocytes; mass-spectrometry; multiple sclerosis; sphingosine kinase; sphingosine 1-phosphate; subcellular compartment

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About the article

Corresponding author: Anja Schwiebs, Pharmazentrum frankfurt/ZAFES, Institute of General Pharmacology and Toxicology, Goethe University Frankfurt, Building 75, Theodor-Stern-Kai 7, D-60590 Frankfurt/Main, Germany, e-mail:

aWe deeply regret the sudden unexpected death of Dr. Helmut Schmidt during the preparation of the manuscript for this article.


Received: 2014-12-01

Accepted: 2015-01-09

Published Online: 2015-01-14

Published in Print: 2015-06-01


Citation Information: Biological Chemistry, Volume 396, Issue 6-7, Pages 795–802, ISSN (Online) 1437-4315, ISSN (Print) 1431-6730, DOI: https://doi.org/10.1515/hsz-2014-0287.

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