Identification of new benzofuran derivatives as STING agonists with broad-spectrum antiviral activity

Virus Res. 2024 Sep:347:199432. doi: 10.1016/j.virusres.2024.199432. Epub 2024 Jul 8.

Abstract

The Stimulator of Interferon Genes (STING) is involved in cytosolic DNA sensing and type I Interferons (IFN-I) induction. Aiming to identify new STING agonists with antiviral activity and given the known biological activity of benzothiazole and benzimidazole derivatives, a series of benzofuran derivatives were tested for their ability to act as STING agonists, induce IFN-I and inhibit viral replication. Compounds were firstly evaluated in a gene reporter assay measuring luciferase activity driven by the human IFN-β promoter in cells expressing exogenous STING (HEK293T). Seven of them were able to induce IFN-β transcription while no induction of the IFN promoter was observed in the presence of a mutated and inactive STING, showing specific protein-ligand interaction. Docking studies were performed to predict their putative binding mode. The best hit compounds were then tested on human coronavirus 229E replication in BEAS-2B and MRC-5 cells and three derivatives showed EC50 values in the μM range. Such compounds were also tested on SARS-CoV-2 replication in BEAS-2B cells and in Calu-3 showing they can inhibit SARS-CoV-2 replication at nanomolar concentrations. To further confirm their IFN-dependent antiviral activity, compounds were tested to verify their effect on phospho-IRF3 nuclear localization, that was found to be induced by benzofuran derivatives, and SARS-CoV-2 replication in Vero E6 cells, lacking IFN production, founding them to be inactive. In conclusion, we identified benzofurans as STING-dependent immunostimulatory compounds and host-targeting inhibitors of coronaviruses representing a novel chemical scaffold for the development of broad-spectrum antivirals.

Keywords: Benzofurans; Broad-spectrum antiviral; Human coronaviruses; Immune modulation; STING.

MeSH terms

  • Animals
  • Antiviral Agents* / chemistry
  • Antiviral Agents* / pharmacology
  • Benzofurans* / chemistry
  • Benzofurans* / pharmacology
  • Cell Line
  • Chlorocebus aethiops
  • HEK293 Cells
  • Humans
  • Interferon-beta / genetics
  • Membrane Proteins* / genetics
  • Membrane Proteins* / metabolism
  • Molecular Docking Simulation
  • SARS-CoV-2 / drug effects
  • Vero Cells
  • Virus Replication* / drug effects

Substances

  • Benzofurans
  • Antiviral Agents
  • STING1 protein, human
  • Membrane Proteins
  • Interferon-beta
  • benzofuran