Targeting Ceramide Metabolism with a Potent and Specific Ceramide Kinase Inhibitor

• Published in: Molecular pharmacology Vol. 74; no. 4; pp. 925 - 932
• Main Authors: Graf, Christine, Klumpp, Martin, Habig, Michael, Rovina, Philipp, Billich, Andreas, Baumruker, Thomas, Oberhauser, Berndt, Bornancin, Frédéric
• Format: Journal Article
• Language: English
• Published: United States American Society for Pharmacology and Experimental Therapeutics 01.10.2008
• Subjects: Animals; Baculoviridae - genetics; Cell Proliferation - drug effects; Cell Survival - drug effects; Cercopithecus aethiops; COS Cells; Dose-Response Relationship, Drug; Glutathione Transferase - metabolism; Humans; Inhibitory Concentration 50; Luciferases - metabolism; Luminescence; Macrophages, Peritoneal - drug effects; Mast Cells - drug effects; Mice; Molecular Structure; Molecular Weight; Phosphorylation - drug effects; Phosphotransferases (Alcohol Group Acceptor) - chemistry; Phosphotransferases (Alcohol Group Acceptor) - genetics; Phosphotransferases (Alcohol Group Acceptor) - isolation & purification; Phosphotransferases (Alcohol Group Acceptor) - metabolism; Protein Kinase Inhibitors - chemistry; Protein Kinase Inhibitors - metabolism; Protein Kinase Inhibitors - pharmacology; Radioligand Assay; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - isolation & purification; Recombinant Fusion Proteins - metabolism; Reproducibility of Results; Sensitivity and Specificity; Spodoptera - cytology; Spodoptera - metabolism; Structure-Activity Relationship
• ISSN: 0026-895X; 1521-0111
• DOI: 10.1124/mol.108.048652

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.