Pharmacological characterization of the seven human NOX isoforms and their inhibitors

Article Article scientifique Article Pharmacological characterization of the seven human NOX isoforms and their inhibitors eng 2019 author Augsburger Fiona 774276 author Filippova Aleksandra 778616 author Rasti-Nadjafabadi Delphine 774398 author Seredenin Tamara 771548 author Lam Magdalena author Maghzal Ghassan author Mahiout Zahia 706663 author Jansen-Dürr Pidder author Knaus Ulla G. author Doroshow James author Stocker Roland 762238 author Krause Karl-Heinz 23022 author Jaquet Vincent 639300 101272 10.1016/j.redox.2019.101272 31330481 2213-2317 https://linkinghub.elsevier.com/retrieve/pii/S2213231719305208 Background NADPH oxidases (NOX) are a family of flavoenzymes that catalyze the formation of superoxide anion radical (O2•-) and/or hydrogen peroxide (H2O2). As major oxidant generators, NOX are associated with oxidative damage in numerous diseases and represent promising drug targets for several pathologies. Various small molecule NOX inhibitors are used in the literature, but their pharmacological characterization is often incomplete in terms of potency, specificity and mode of action. Experimental approach We used cell lines expressing high levels of human NOX isoforms (NOX1-5, DUOX1 and 2) to detect NOX-derived O2•- or H2O2 using a variety of specific probes. NOX inhibitory activity of diphenylene iodonium (DPI), apocynin, diapocynin, ebselen, GKT136901 and VAS2870 was tested on NOX isoforms in cellular and membrane assays. Additional assays were used to identify potential off target effects, such as antioxidant activity, interference with assays or acute cytotoxicity. Key results Cells expressing active NOX isoforms formed O2•-, except for DUOX1 and 2, and in all cases activation of NOX isoforms was associated with the detection of extracellular H2O2. Among all molecules tested, DPI elicited dose-dependent inhibition of all isoforms in all assays, however all other molecules tested displayed interesting pharmacological characteristics, but did not meet criteria for bona fide NOX inhibitors. Conclusion Our findings indicate that experimental results obtained with widely used NOX inhibitors must be carefully interpreted and highlight the challenge of developing reliable pharmacological inhibitors of these key molecular targets. NADPH oxidase NOX Reactive oxygen species Small molecule inhibitors Dewey 616.07 European Commission NOX enzymes as mediators of inflammation-triggered neurodegeneration: modulating NOX enzymes as novel therapies 278611 NEURINOX EU FP7 137 Département de pathologie et immunologie 21F false 38 Radicaux libres et cellules souches embryonnaires 60 Redox Biology 26 052b8e89-6bd1-494e-85f8-762c1cb028a0 Article (Published version) Public 1-s2.0-S2213231719305208-main.pdf 2732279 application/pdf CC BY openAccess Faculté de médecine / Section de médecine fondamentale / Département de pathologie et immunologie

Title	Published in	Year	Views	Downloads

NOX5 cell-free assay for the high-throughput screening of small molecules	Methods in Molecular Biology	2019	406	3
Pharmacological characterization of the seven human NOX isoforms and their inhibitors	Redox Biology	2019	451	430

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Rasti-Nadjafabadi, Delphine