Structure-based discovery of conformationally selective inhibitors of the serotonin transporter

The serotonin transporter (SERT) removes synaptic serotonin and is the target of anti-depressant drugs. SERT adopts three conformations: outward-open, occluded, and inward-open. All known inhibitors target the outward-open state except ibogaine, which has unusual anti-depressant and substance-withdrawal effects, and stabilizes the inward-open conformation. Unfortunately, ibogaine’s promiscuity and cardiotoxicity limit the understanding of inward-open state ligands. We docked over 200 million small molecules against the inward-open state of the SERT. Thirty-six top-ranking compounds were synthesized, and thirteen inhibited; further structure-based optimization led to the selection of two potent (low nanomolar) inhibitors. These stabilized an outward-closed state of the SERT with little activity against common off-targets. A cryo-EM structure of one of these bound to the SERT confirmed the predicted geometry. In mouse behavioral assays, both compounds had anxiolytic- and anti-depressant-like activity, with potencies up to 200-fold better than fluoxetine (Prozac), and one substantially reversed morphine withdrawal effects. [Display omitted] •Large-library docking finds conformationally and target-selective SERT inhibitors•Cryo-EM supports the computationally predicted structure•The inhibitors are anti-depressant-like and alleviate opioid withdrawal in mice•Targeting of transporters for structure-based ligand discovery The development of low nM inhibitors that confer anti-depressant-like and anti-opioid withdrawal actions in mice was achieved by screening a large virtual library against the inward-open conformation of SERT.