TY  - JOUR
A1  - Imeri, Faik
A1  - Fallegger, Daniel
A1  - Zivkovic, Aleksandra
A1  - Schwalm, Stephanie
A1  - Enzmann, Gaby
A1  - Blankenbach, Kira
A1  - Heringdorf, Dagmar Meyer Zu
A1  - Homann, Thomas
A1  - Kleuser, Burkhard
A1  - Pfeilschifter, Josef
A1  - Engelhardt, Britta
A1  - Stark, Holger
A1  - Huwiler, Andrea
T1  - Novel oxazolo-oxazole derivatives of FTY720 reduce endothelial cell permeability, immune cell chemotaxis and symptoms of experimental autoimmune encephalomyelitis in mice
JF  - Neuropharmacology
N2  - The immunomodulatory FTY720 (fingolimod) is presently approved for the treatment of relapsing-remitting multiple sclerosis. It is a prodrug that acts by modulating sphingosine 1-phosphate (S1P) receptor signaling. In this study, we have developed and characterized two novel oxazolo-oxazole derivatives of FTY720, ST-968 and the oxy analog ST-1071, which require no preceding activating phosphorylation, and proved to be active in intact cells and triggered S1P(1) and S1P(3), but not S1P(2), receptor internalization as a result of receptor activation.
 Functionally, ST-968 and ST-1071 acted similar to FTY720 to abrogate S1P-triggered chemotaxis of mouse splenocytes, mouse T cells and human U937 cells, and reduced TNFa- and LPS-stimulated endothelial cell permeability. The compounds also reduced TNF alpha-induced ICAM-1 and VCAM-1 mRNA expression, but restored TNF alpha-mediated downregulation of PECAM-1 mRNA expression.
 In an in vivo setting, the application of ST-968 or ST-1071 to mice resulted in a reduction of blood lymphocytes and significantly reduced the clinical symptoms of experimental autoimmune encephalomyelitis (EAE) in C57BL/6 mice comparable to FTY720 either by prophylactic or therapeutic treatment. In parallel to the reduced clinical symptoms, infiltration of immune cells in the brain was strongly reduced, and in isolated tissues of brain and spinal cord, the mRNA and protein expressions of ICAM-1 and VCAM-1, as well as of matrix metalloproteinase-9 were reduced by all compounds, whereas PECAM-1 and tissue inhibitor of metalloproteinase TIMP-1 were upregulated.
 In summary, the data suggest that these novel butterfly derivatives of FTY720 could have considerable implication for future therapies of multiple sclerosis and other autoimmune diseases. (C) 2014 Elsevier Ltd. All rights reserved.
KW  - Fingolimod
KW  - ST-968
KW  - ST-1071
KW  - Sphingosine 1-phosphate
KW  - Endothelial cells
KW  - Permeability
KW  - Multiple sclerosis
Y1  - 2014
U6  - https://doi.org/10.1016/j.neuropharm.2014.05.012
SN  - 0028-3908
SN  - 1873-7064
VL  - 85
SP  - 314
EP  - 327
PB  - Elsevier
CY  - Oxford
ER  - 
TY  - JOUR
A1  - Pastukhov, Oleksandr
A1  - Schwalm, Stephanie
A1  - Zangemeister-Wittke, Uwe
A1  - Fabbro, Doriano
A1  - Bornancin, Frederic
A1  - Japtok, Lukasz
A1  - Kleuser, Burkhard
A1  - Pfeilschifter, Josef
A1  - Huwiler, Andrea
T1  - The ceramide kinase inhibitor NVP-231 inhibits breast and lung cancer cell proliferation by inducing M phase arrest and subsequent cell death
JF  - British journal of pharmacology : journal of The British Pharmacological Society
N2  - Background and PurposeCeramide kinase (CerK) catalyzes the generation of ceramide-1-phosphate which may regulate various cellular functions, including inflammatory reactions and cell growth. Here, we studied the effect of a recently developed CerK inhibitor, NVP-231, on cancer cell proliferation and viability and investigated the role of cell cycle regulators implicated in these responses.
 Experimental ApproachThe breast and lung cancer cell lines MCF-7 and NCI-H358 were treated with increasing concentrations of NVP-231 and DNA synthesis, colony formation and cell death were determined. Flow cytometry was performed to analyse cell cycle distribution of cells and Western blot analysis was used to detect changes in cell cycle regulator expression and activation.
 Key ResultsIn both cell lines, NVP-231 concentration-dependently reduced cell viability, DNA synthesis and colony formation. Moreover it induced apoptosis, as measured by increased DNA fragmentation and caspase-3 and caspase-9 cleavage. Cell cycle analysis revealed that NVP-231 decreased the number of cells in S phase and induced M phase arrest with an increased mitotic index, as determined by increased histone H3 phosphorylation. The effect on the cell cycle was even more pronounced when NVP-231 treatment was combined with staurosporine. Finally, overexpression of CerK protected, whereas down-regulation of CerK with siRNA sensitized, cells for staurosporine-induced apoptosis.
 Conclusions and ImplicationsOur data demonstrate for the first time a crucial role for CerK in the M phase control in cancer cells and suggest its targeted inhibition, using drugs such as NVP-231, in combination with conventional pro-apoptotic chemotherapy.
Y1  - 2014
U6  - https://doi.org/10.1111/bph.12886
SN  - 0007-1188
SN  - 1476-5381
VL  - 171
IS  - 24
SP  - 5829
EP  - 5844
PB  - Wiley-Blackwell
CY  - Hoboken
ER  -