A Putative Frizzled 7-Targeting Compound Acts as a Firefly Luciferase Inhibitor

作者：Julia Kinsolving, Lukas Grätz, Jan Voss, Bente Löw, Emily Shorter, Baptiste Jude, Johanna T. Lanner, Stefan Löber, Peter Gmeiner, Gunnar Schulte · 发表于：Journal of Medicinal Chemistry · 年份：2024 · DOI：10.1021/acs.jmedchem.4c02766 · 被引用次数：7 · 研究领域：bioluminescence and chemiluminescence research、Neurobiology and Insect Physiology Research、Nicotinic Acetylcholine Receptors Study

High Resolution Image Download MS PowerPoint Slide The Frizzled family (FZD 1–10 ) of G protein-coupled receptors regulates WNT signaling mediating proliferative input. Dysregulation of FZD 7 and exaggerated WNT/β-catenin signaling is frequently observed in intestinal cancers. Therefore, it is attractive to develop therapeutics targeting FZD 7 for cancer treatment. Structure-based virtual screening has identified compound 28, which inhibited WNT/β-catenin signaling based on the luciferase-based reporter gene TOPFlash assay. However, upon pharmacological validation, compound 28 rather acts as a potent Firefly luciferase (Fluc) inhibitor (IC 50 = 30 nM), matching the reported IC 50 for compound 28-mediated inhibition in the TOPFlash assay. Moreover, we employed Fluc-independent assays, a FZD 7 -focused bioluminescence resonance energy transfer biosensor and quantitative PCR, to emphasize the inability of compound 28 to inhibit the WNT-3A-induced conformational dynamics in FZD 7 and transcription of Axin2, a WNT target gene. Thus, we underline the importance of counter screens to validate compounds that interfere with the detection technology used for compound screening.