Abstract
Ceramide kinase (CerK) produces the bioactive lipid ceramide-1-phosphate (C1P) and appears as a key enzyme for controlling ceramide levels. In this study, we discovered and characterized adamantane-1-carboxylic acid (2-benzoylamino-benzothiazol-6-yl)amide (NVP-231), a potent, specific, and reversible CerK inhibitor that competitively inhibits binding of ceramide to CerK. NVP-231 is active in the low nanomolar range on purified as well as cellular CerK and abrogates phosphorylation of ceramide, resulting in decreased endogenous C1P levels. When combined with another ceramide metabolizing inhibitor, such as tamoxifen, NVP-231 synergistically increased ceramide levels and reduced cell growth. Therefore, NVP-231 represents a novel and promising compound for controlling ceramide metabolism that may provide insight into CerK physiological function.
Footnotes
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ABBREVIATIONS: CerK, ceramide kinase; C1P, ceramide-1-phosphate; MOPS, 3-(N-morpholino)propanesulfonic acid; DMSO, dimethyl sulfoxide; TLC, thin-layer chromatography; LC/MS, liquid chromatography/mass spectrometry; BMDM, bone marrow-derived macrophages; 7-AAD, 7-aminoactinomycin D; GlcCer, glucosylceramide; SM, sphingomyelin; M-CSF, macrophage colony stimulating factor; PLA2G4A, cytosolic phospholipase A2 α; NBD, N-[7-(4-nitrobenzo-2-oxa-l,3-diazole)]; NBD-Cer, NBD-C6-ceramide; NVP-231, adamantane-1-carboxylic acid (2-benzoylamino-benzothiazol-6-yl)amide.
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The online version of this article (available at http://molpharm.aspetjournals.org) contains supplemental material. - Received May 6, 2008.
- Accepted July 1, 2008.
- The American Society for Pharmacology and Experimental Therapeutics
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