New chemical tools to illuminate N-acylphosphatidylethanolamine biosynthesis

Wendel, T.J. (2023)

Doctoral Thesis

This thesis describes the development and optimization of the first molecular tools to study the enzyme PLA2G4E. After remaining elusive for many years, in 2016, this enzyme was discovered to be responsible for the calcium-dependent formation of N-acylphosphatidylethanolamines (NAPEs) in cells. NAPEs are low-abundant lipid species that play roles in membrane stabilization, cell signaling and homeostasis. They are well-known as precursors to the signaling lipids of the N-acylethanolamine (NAE) class, but their own biological functions remain relatively poorly understood. To find inhibitors of PLA2G4E, a focused compound library was screened in a newly developed activity-based protein profiling (ABPP) assay. Hits were identified and optimized by building structure-activity relationships (SARs) through organic synthesis and activity assays. WEN091 was identified as potent inhibitor of PLA2G4E that was able to reduce cellular NAPE levels. Cellular target engagement was confirmed by use of a tailored activity-based probe. Using these molecular tools, the relevance of NAPEs and PLA2G4E in cellular processes and disease may be elucidated.

• Chemical biology
• Medicinal chemistry
• Enzyme inhibitors
• Activity-based protein profiling
• Endocannabinoid system
• N-acylphosphatidylethanolamine
• N-acyltransferase
• 2-arachidonoylglycerol
• PLA2G4E
• ABHD6

• All authors: Wendel, T.J.
• Supervisor: Stelt, M. van der; Boeckel, C.A.A. van
• Co-supervisor: Berg, R.J.B.H.N. van den
• Committee: Aerts, J.M.F.G.; Overkleeft, H.S.; Artola Pérez de Azanza, M.E.; Zimmermann, R.; Verhelst, S.H.L.
• Qualification: Doctor (dr.)
• Awarding Institution: Leiden Institute of Chemistry (LIC), Faculty of Science, Leiden University
• Date: 2023-03-23
• ISBN (print): 9789464730609