Next-generation small molecule inhibitors of clathrin function acutely inhibit endocytosis

  • Structure. 2025 May 1;33(5):878-890.e7. doi: 10.1016/j.str.2025.02.011.
André Horatscheck  1 ,  Michael Krauß  1 ,  Haydar Bulut  2 ,  Valerie Chambon  3 ,  Massilullah Shafaq Zadah  3 ,  Estelle Dransart  3 ,  Kimberly Peloza  1 ,  Karine F Santos  4 ,  Mark J Robertson  5 ,  Kate Prichard  5 ,  Sandra Miksche  1 ,  Silke Radetzki  1 ,  Jens-Peter von Kries  1 ,  Markus C Wahl  6 ,  Adam McCluskey  5 ,  Ludger Johannes  3 ,  Volker Haucke  7 ,  Marc Nazaré  8
Affiliations
  • 1. Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, 13125 Berlin, Germany.
  • 2. Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, 13125 Berlin, Germany; Department of Biology, Chemistry, Pharmacy, Freie Universität Berlin, 14195 Berlin, Germany.
  • 3. Institut Curie, PSL Research University, Cellular and Chemical Biology Unit, U1143 INSERM, UMR3666 CNRS, 26 rue d'Ulm, 75248 Paris Cedex 05, France.
  • 4. Department of Biology, Chemistry, Pharmacy, Freie Universität Berlin, 14195 Berlin, Germany.
  • 5. Chemistry, School of Environmental & Life Sciences, The University of Newcastle, Callaghan, NSW 2308, Australia.
  • 6. Department of Biology, Chemistry, Pharmacy, Freie Universität Berlin, 14195 Berlin, Germany; Helmholtz-Zentrum Berlin für Materialien und Energie, Macromolecular Crystallography, Albert-Einstein-Str. 15, 12489 Berlin, Germany.
  • 7. Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, 13125 Berlin, Germany; Department of Biology, Chemistry, Pharmacy, Freie Universität Berlin, 14195 Berlin, Germany. Electronic address: [email protected].
  • 8. Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, 13125 Berlin, Germany; Helmholtz-Zentrum Berlin für Materialien und Energie, Macromolecular Crystallography, Albert-Einstein-Str. 15, 12489 Berlin, Germany. Electronic address: [email protected].
PMID: 40112806 DOI: 10.1016/j.str.2025.02.011
Abstract

Clathrin-mediated endocytosis (CME) is the predominant endocytic pathway in eukaryotic cells and a major regulator of cell physiology as it facilitates the internalization of receptors, channels, and transporters and viral entry. The clathrin terminal domain acts as a central protein interaction hub within the endocytic protein network. Previously described inhibitors of CME display off-target activities that result in cytotoxicity, providing limitations to their use. We report the development and characterization of next-generation small molecule inhibitors of clathrin terminal domain function. These compounds termed Pitstop 2c and Pitstop 2d occupy the binding site within the clathrin terminal domain for endocytic protein ligands including epsin, resulting in potent inhibition of receptor-mediated endocytosis and reduced entry of vesicular stomatitis virus (VSV) with minimal cytotoxic side effects. Next-generation Pitstops thus provide an improved toolset to address clathrin function in cell physiology with potential applications as inhibitors of virus and pathogen entry.

Keywords
cathrin; endocytosis; small molecule inhibition; virus entry.
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