Selective covalent targeting of GPX4 using masked nitrile-oxide electrophiles

We recently described glutathione peroxidase 4 (GPX4) as a promising target for killing therapy-resistant cancer cells via ferroptosis. The onset of therapy resistance by multiple types of treatment results in a stable cell state marked by high levels of polyunsaturated lipids and an acquired depend...

Full description

Saved in:
Bibliographic Details
Published inNature chemical biology Vol. 16; no. 5; pp. 497 - 506
Main Authors Eaton, John K., Furst, Laura, Ruberto, Richard A., Moosmayer, Dieter, Hilpmann, André, Ryan, Matthew J., Zimmermann, Katja, Cai, Luke L., Niehues, Michael, Badock, Volker, Kramm, Anneke, Chen, Sixun, Hillig, Roman C., Clemons, Paul A., Gradl, Stefan, Montagnon, Claire, Lazarski, Kiel E., Christian, Sven, Bajrami, Besnik, Neuhaus, Roland, Eheim, Ashley L., Viswanathan, Vasanthi S., Schreiber, Stuart L.
Format Journal Article
LanguageEnglish
Published New York Nature Publishing Group US 01.05.2020
Nature Publishing Group
Subjects
Online AccessGet full text
ISSN1552-4450
1552-4469
1552-4469
DOI10.1038/s41589-020-0501-5

Cover

More Information
Summary:We recently described glutathione peroxidase 4 (GPX4) as a promising target for killing therapy-resistant cancer cells via ferroptosis. The onset of therapy resistance by multiple types of treatment results in a stable cell state marked by high levels of polyunsaturated lipids and an acquired dependency on GPX4. Unfortunately, all existing inhibitors of GPX4 act covalently via a reactive alkyl chloride moiety that confers poor selectivity and pharmacokinetic properties. Here, we report our discovery that masked nitrile-oxide electrophiles, which have not been explored previously as covalent cellular probes, undergo remarkable chemical transformations in cells and provide an effective strategy for selective targeting of GPX4. The new GPX4-inhibiting compounds we describe exhibit unexpected proteome-wide selectivity and, in some instances, vastly improved physiochemical and pharmacokinetic properties compared to existing chloroacetamide-based GPX4 inhibitors. These features make them superior tool compounds for biological interrogation of ferroptosis and constitute starting points for development of improved inhibitors of GPX4. Nitrile-oxide electrophiles were identified as covalent inhibitors of GPX4 that exhibit increased selectivity and reduced off-target effects relative to chloroacetamide-based inhibitors.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 14
content type line 23
AUTHOR CONTRIBUTIONS
J.K.E. conceived and designed experiments. J.K.E., L.F., K.E.L., S.G., and C.M. performed chemical synthesis and compound characterization. J.K.E., R.A.R., M.J.R., L.L.C., and V.S.V. maintained cell cultures and performed viability, cellular thermal shift, and western blotting experiments. D.M. and A.H. designed the cloning approach and expressed, purified, and characterized recombinant wild-type GPX4 protein. V.B., A.H., D.M., and J.K.E. performed cellular and biochemical MS binding assays. M.N. performed metabolite-ID studies. R.C.H., K.Z., A.K., S.C., B.B. contributed tools and reagents for protein characterization experiments. P.A.C. contributed to analysis of cell viability data. R.A.R. and S.Ch. performed cellular lipid peroxidation assays. R.N. performed formulation work. A.L.E. performed in vivo experiments. J.K.E., V.S.V. and S.L.S. initiated the project and wrote the manuscript. V.S.V. and S.L.S. directed the project.
ISSN:1552-4450
1552-4469
1552-4469
DOI:10.1038/s41589-020-0501-5