Identification of Three Antimicrobials Activating Serotonin Receptor 4 in Colon Cells

[Image: see text] The serotonin receptor 4b (5-HTR(4b)) is expressed throughout the gastrointestinal tract, and its agonists are used in the treatment of irritable bowel syndrome with constipation (IBS-C). Today, there are no rapid assays for the identification of 5-HTR(4b) agonists. Here, we developed a luciferase-based 5-HTR(4b) assay capable of assessing one compound per second with a 38-fold dynamic range and nM limit of detection for serotonin. We used the assay to screen more than 1000 natural products and anti-infection agents and identified five new 5-HTR(4b) ligands: hordenine, halofuginone, proflavine, ethacridine, and revaprazan. We demonstrate that hordenine (antibiofilm), halofuginone (antiparasitic), and revaprazan (gastric acid reducer) activate 5-HTR(4b) in human colon epithelial cells, leading to increased cell motility or wound healing. The 5-HTR(4b) assay can be used to screen larger pharmaceutical libraries to identify novel treatments for IBS-C. This work shows that antimicrobials interact not only with the gut microbiota, but also with the human host.