High-throughput discovery of novel small-molecule inhibitors of acid Ceramidase

Abstract

Ceramide has a key role in the regulation of cellular senescence and apoptosis. As Ceramide levels are lowered by the action of acid ceramidase (AC), abnormally expressed in various cancers, the identification of AC inhibitors has attracted increasing interest. However, this finding has been mainly hampered by the lack of formats suitable for the screening of large libraries. We have overcome this drawback by adapting a fluorogenic assay to a 384-well plate format. The performance of this optimised platform has been proven by the screening a library of 4100 compounds. Our results show that the miniaturised platform is well suited for screening purposes and it led to the identification of several hits, that belong to different chemical classes and display potency ranges of 2–25 µM. The inhibitors also show selectivity over neutral ceramidase and retain activity in cells and can therefore serve as a basis for further chemical optimisation.

Keywords:

• Sphingolipids
• ceramide
• acid ceramidase
• inhibitor

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work was supported by the Ministerio de Economica, Industria y Competitividad under the Grant CTQ2017-85378-R. The authors thank Eli Lilly for providing the compounds tested in this study. Mazen Aseeri was financially supported by a scholarship from the Saudi Arabian Ministry of Higher Education.