Laudanosine restricts Ebola virus entry by targeting TPC2-dependent endolysosomal trafficking

作者：Tom Seitz, Julian Koengeter, Susanne Klute, Franziska Kraft, Denise Christine Clesle, Lily Tschampel, Nico Preising, Armando Alexei Rodríguez Alfonso, Sebastian Wiese, Ludger Ständker, Christoph Jung, Timo Jacob, Janet Köhler, Gilbert Weidinger, Nadine Biedenkopf, Konstantin M. J. Sparrer, Frank Kirchhoff, Fabian Zech · 发表于：bioRxiv (Cold Spring Harbor Laboratory) · 年份：2026 · DOI：10.64898/2026.06.17.732874 · 研究领域：Viral Infections and Outbreaks Research、Calcium signaling and nucleotide metabolism、Cellular transport and secretion

ABSTRACT Late endosome-dependent viruses, including filo- and arenaviruses, rely on host endolysosomal trafficking for productive infection. Here, we used a dual-colour Vesicular stomatitis virus (VSV) based pseudoparticle screen of CytoSorb-derived fractions to identify inhibitors of the Zaire Ebolavirus glycoprotein (GP)-mediated entry. Iterative chromatographic purification and mass spectrometry identified Laudanosine, a degradation product of the clinically used neuromuscular blocker Atracurium, as the antiviral compound. Laudanosine specifically inhibited entry mediated by Ebola, Marburg, Lymphocytic choriomeningitis and Lassa virus glycoproteins without affecting VSV-G-dependent entry. Importantly, Laudanosine inhibited authentic Ebola virus infection without detectable cytotoxicity in cell culture and embryonic zebrafish. Molecular dynamics simulations suggest stable association of Laudanosine with the allosteric inhibitory pocket of the lysosomal two-pore channel (TPC2). Consistently, Laudanosine impairs autophagic flux and disrupts endolysosomal trafficking. Together, our findings identify Laudanosine as a previously unrecognised inhibitor of TPC2-dependent entry of highly lethal viral pathogens.