Structure-Based Design of a Chemical Probe Set for the 5-HT 5A Serotonin Receptor.

The 5-HT receptor (5-HTR), for which no selective agonists and a few antagonists exist, remains the least understood serotonin receptor. A single commercial antagonist, SB-699551, has been widely used to investigate the 5-HTR function in neurological disorders, including pain, but this molecule has substantial liabilities as a chemical probe. Accordingly, we sought to develop an internally controlled probe set. Docking over 6 million molecules against a 5-HTR homology model identified 5 mid-μM ligands, one of which was optimized to , a 42 nM arrestin-biased partial agonist at the 5-HTR with a more restricted off-target profile and decreased assay liabilities versus SB-699551. Site-directed mutagenesis supported the docked pose of . Surprisingly, analogs of that lost the 5-HTR activity revealed that 5-HTR engagement is nonessential for alleviating pain, contrary to studies with less-selective ligands. and analogs constitute a selective probe set with which to study the function of the 5-HTR.