title: Targeting of Helicobacter pylori thymidylate synthase ThyX by non-mitotoxic hydroxy-naphthoquinones.
abstract: ThyX is an essential thymidylate synthase that is mechanistically and structurally unrelated to the functionally analogous human enzyme, thus providing means for selective inhibition of bacterial growth. To identify novel compounds with anti-bacterial activity against the human pathogenic bacterium Helicobacter pylori, based on our earlier biochemical and structural analyses, we designed a series of eighteen 2-hydroxy-1,4-naphthoquinones (2-OH-1,4-NQs) that target HpThyX. Our lead-like molecules markedly inhibited the NADPH oxidation and 2'-deoxythymidine-5'-monophosphate-forming activities of HpThyX enzyme in vitro, with inhibitory constants in the low nanomolar range. The identification of non-cytotoxic and non-mitotoxic 2-OH-1,4-NQ inhibitors permitted testing their in vivo efficacy in a mouse model for H. pylori infections. Despite the widely assumed toxicity of naphthoquinones (NQs), we identified tight-binding ThyX inhibitors that were tolerated in mice and can be associated with a modest effect in reducing the number of colonizing bacteria. Our results thus provide proof-of-concept that targeting ThyX enzymes is a highly feasible strategy for the development of therapies against H. pylori and a high number of other ThyX-dependent pathogenic bacteria. We also demonstrate that chemical reactivity of NQs does not prevent their exploitation as anti-microbial compounds, particularly when mitotoxicity screening is used to prioritize these compounds for further experimentation.
authors.affiliation: Laboratoire d'Optique et Biosciences, CNRS UMR7645, INSERM U1182, Ecole Polytechnique, Palaiseau 91128, France Department of Biology, Université Paris-Sud, Orsay 91405, France stephane.skouloubris@u-psud.fr.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Skouloubris
authors.affiliation: Laboratoire d'Optique et Biosciences, CNRS UMR7645, INSERM U1182, Ecole Polytechnique, Palaiseau 91128, France.
authors.forename: Kamel
authors.initials: K
authors.lastname: Djaout
authors.affiliation: Laboratoire d'Optique et Biosciences, CNRS UMR7645, INSERM U1182, Ecole Polytechnique, Palaiseau 91128, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Lamarre
authors.affiliation: Laboratoire d'Optique et Biosciences, CNRS UMR7645, INSERM U1182, Ecole Polytechnique, Palaiseau 91128, France.
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Lambry
authors.affiliation: Department of Microbiology, Institut Pasteur, Unité Pathogenèse de Helicobacter, 28 rue du Dr. Roux, Paris 75724, France.
authors.forename: Karine
authors.initials: K
authors.lastname: Anger
authors.affiliation: Laboratoire d'Optique et Biosciences, CNRS UMR7645, INSERM U1182, Ecole Polytechnique, Palaiseau 91128, France.
authors.forename: Julien
authors.initials: J
authors.lastname: Briffotaux
authors.affiliation: Laboratoire d'Optique et Biosciences, CNRS UMR7645, INSERM U1182, Ecole Polytechnique, Palaiseau 91128, France.
authors.forename: Ursula
authors.initials: U
authors.lastname: Liebl
authors.affiliation: Department of Microbiology, Institut Pasteur, Unité Pathogenèse de Helicobacter, 28 rue du Dr. Roux, Paris 75724, France.
authors.forename: Hilde
authors.initials: H
authors.lastname: de Reuse
authors.affiliation: Laboratoire d'Optique et Biosciences, CNRS UMR7645, INSERM U1182, Ecole Polytechnique, Palaiseau 91128, France hannu.myllykallio@polytechnique.edu.
authors.forename: Hannu
authors.initials: H
authors.lastname: Myllykallio
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D005434:Flow Cytometry; D016481:Helicobacter Infections / Q000188:drug therapy* / Q000201:enzymology / Q000382:microbiology; D016480:Helicobacter pylori / Q000187:drug effects* / Q000201:enzymology; D006801:Humans; D051379:Mice; D008938:Mitosis / Q000187:drug effects; D009285:Naphthoquinones / Q000737:chemistry / Q000494:pharmacology*; D013274:Stomach Neoplasms / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology*; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors*; D014407:Tumor Cells, Cultured
chemical_list: D004791:Enzyme Inhibitors; D009285:Naphthoquinones; D013940:Thymidylate Synthase
keywords: Helicobacter pylori; anti-microbial agents; naphthoquinone; thymidylate synthase ThyX