Targeting Ceramide Metabolism with a Potent and Specific Ceramide Kinase Inhibitor

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.

Characterization of eicosanoid synthesis in a genetic ablation model of ceramide kinase

“…This further illustrates the effects of CERK-derived C1P on eicosanoid production, and therefore implicates these mediators as players in the infl ammatory response. Of note, these fi ndings are partially in opposition to the reports by Bornancin and coworkers ( 23,24,49 ). Specifi cally, this group explored how cPLA 2 ␣ -dependent pathways were affected by the genetic ablation of CERK and this laboratory observed similar responses of cells derived from both wild-type and knockout animals to PMA/ionomycin treatment as well as the susceptibility of the whole animals to arthritis induction by both Ag and KRN serum.…”

“…This may be one alternative route to determining the role of a particular bioactive lipid in a particular biology to overcome compensatory mechanisms when targeting the biosynthetic pathways. As stated previously, there is still the question of the CERK inhibitor (NVP-231) developed by Novartis ( 49 ). Bornancin and coworkers reported that treatment of cells with this inhibitor had no effect on the stimulation of eicosanoid synthesis by various infl ammatory agonists ( 49 ).…”

Activation of Ceramidase and Ceramide Kinase by Vanadate via a Tyrosine Kinase–Mediated Pathway

“…In the present study, the expression of hCERK increased the level of NBD-C1P accompanied with decreases in NBD-SM and NBD-GlcCer in CHO cells (Table 2). Similarly, Graf et al (14) reported that expression of CERK reduced levels of NBD-SM and NBD-GlcCer. Inhibition of NBD-GlcCer synthesis by PPMP in the present study and by tamoxifen (14) enhanced the formation of NBD-SM in CHO cells and in COS cells, respectively.…”

“…Similarly, Graf et al (14) reported that expression of CERK reduced levels of NBD-SM and NBD-GlcCer. Inhibition of NBD-GlcCer synthesis by PPMP in the present study and by tamoxifen (14) enhanced the formation of NBD-SM in CHO cells and in COS cells, respectively. MAPPsensitive alkaline ceramidases such as ACER2 and ACER3 are localized to the Golgi apparatus (39,49).…”

Featuring… Susumu Mitsutake

“…A specific inhibitor for CERK has already been developed [5], so we are focusing on the SMS2 inhibitor. We have already patented a screening method for discriminating between SMS1 and SMS2, which we expect to be helpful for identifying an SMS2-specific inhibitor.…”