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US 5,851,782 AGranted
Inhibitors of ceramidase
AssigneeDuke University
InventorsYusuf A. Hannun, Alicja Bielawska, Charles McKay
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Abstract
The present invention relates, in general, to the bioeffector molecule ceramide and, in particular, to methods of effecting intracellular accumulation of ceramide. The invention further relates to methods of selecting compounds that inhibit alkaline ceramidase and that can be used to treat diseases/disorders associated with cell hyperplasia or dedifferentiation.
Claims4
A method of testing a compound for its ability to ilhibit mammalian alkaline ceramidase comprising
- i) preparing a reaction mixture comprising said compound, mammalian alkaline ceramidase and ceramide, and incubating said mixture under conditions such that said alkaline ceramidase degrades ceramide to sphingosine in the absence of said compound, and
- ii) measuring the level of degradation of said ceramide in the presence of said compound and comparing that level to a level of degradation of ceramide obtained in the absence of said compound
- wherein a lower level of degradation of ceramide in the prcesence of said compound indicates that said compound inhibits said alkaline ceramidase.
A method of testing a compound for its ability to inhibit mammalian alkaline ceramidase comprising
- i) preparing reaction mixtures comprising mammalian alkaline ceramidase, ceramide, and differing concentrations of a known inhibitor of said alkaline ceramidase,
- ii) incubating said mixtures under conditions such that said alkaline cerimidase degrades ceramide to sphingosine in the absence of said known inhibitor, and measuring the effect of said differing concentrations of said known inhibitor on degradation of said ceramide by said alkaline ceramidase so that a dose response curve for said known inhibitor is obtained, and
- iii) measuring the effect of said compound on said dose response curve of said known inhibitor, wherein a shift of said dose response curve of said known inhibitor to the left by said compound indicates that said compound inhibits alkaline cerarnidase.
Dependencies are inferred from the claim text.