Biological and Medicinal Chemistry

Development of a NanoBRET assay to validate inhibitors of Sirt2-mediated lysine deacetylation and defatty-acylation that block prostate cancer cell migration

Authors

  • Anja Vogelmann Institute of Pharmaceutical Sciences & University of Freiburg ,
  • Matthias Schiedel University of Erlangen-Nuremberg & Department of Chemistry and Pharmacy, Medicinal Chemistry ,
  • Nathalie Wössner Institute of Pharmaceutical Sciences & University of Freiburg ,
  • Annika Merz Institute of Pharmaceutical Sciences & University of Freiburg ,
  • Daniel Herp Institute of Pharmaceutical Sciences & University of Freiburg ,
  • Sören Hammelmann Institute of Pharmaceutical Sciences & University of Freiburg ,
  • Arianna Colcerasa Institute of Pharmaceutical Sciences & University of Freiburg ,
  • Garrison Komaniecki Department of Chemistry and Chemical Biology & Cornell University ,
  • Jun Young Hong Department of Chemistry and Chemical Biology & Cornell University ,
  • Manuela Sum Department of Urology and Center for Clinical Research & University Medical Center Freiburg ,
  • Eric Metzger Department of Urology and Center for Clinical Research & University Medical Center Freiburg ,
  • Emilia Neuwirt Institute of Neuropathology & University Medical Center Freiburg ,
  • Lin Zhang Institute of Biochemistry & University of Freiburg ,
  • Oliver Einsle Institute of Biochemistry & University of Freiburg ,
  • Olaf Groß Institute of Neuropathology & University Medical Center Freiburg ,
  • Roland Schüle Department of Urology and Center for Clinical Research & University Medical Center Freiburg ,
  • Hening Lin Cornell University & Department of Chemistry and Chemical Biology & Howard Hughes Medical Institute ,
  • Wolfgang Sippl Department of Medicinal Chemistry, Institute of Pharmacy & Martin Luther University Halle-Wittenberg ,
  • Manfred Jung Institute of Pharmaceutical Sciences, University of Freiburg

Abstract

Sirtuin2 (Sirt2) with its NAD+-dependent deacetylase and defatty-acylase activities plays a central role in the regulation of specific cellular functions. Dysregulation of Sirt2 activity has been associated with the pathogenesis of many diseases, thus making Sirt2 a promising target for pharmaceutical intervention. Herein, we present new high affinity Sirt2 selective Sirtuin-Rearranging Ligands (SirReals) that inhibit both Sirt2-dependent deacetylation and defatty-acylation in vitro and in cells. We show that simultaneous inhibition of both Sirt2 activities results in strongly reduced levels of the oncogene c-Myc and an inhibition of cancer cell migration. Furthermore, we describe the development of a NanoBRET-based assay for Sirt2, thereby providing a method to study cellular target engagement for Sirt2 in a straightforward and accurately quantifiable manner. Applying this assay, we could confirm cellular Sirt2 binding of our new Sirt2 inhibitors and correlate their anticancer effects with their cellular target engagement.

Version notes

Added biochemical data and cellular data for selected inhibitors

Content

Thumbnail image of Sirt2_paper_main_text_23022022_chemrxiv.pdf

Supplementary material

Thumbnail image of Sirt2_paper_ESI_23022022.pdf
Development of a NanoBRET assay to validate inhibitors of Sirt2-mediated lysine deacetylation and defatty-acylation that block prostate cancer cell migration
Supplementary material: additional cellular data, docking study, synthesis procedures, western blot raw data