Fig. 3. (a) Hit validation with the affinity MS assay on a mixture of the 11 putative ligands. Confirmed ligands (mean BI > 2 and p < 0.01) are designated by asterisks, with data shown as mean and s.d. of four individual assays. (b) Affinity of 3943 to 5HT2CR determined by radioligand binding assay. (c) Inhibition of 5HT2CR activation by 3943 revealed by calcium mobilization assay. MES, mesulergine. Ki and IC50 measurement in (b) and (c) were represented by mean and s.d. from experimental triplicates. (d) Docked poses of 3943 (blue) in the 5-HT2CR model (yellow). (e) Schematic of the interactions between 3943 and 5-HT2CR. Residue colors: negatively charged, red; hydrophobic/aromatic, green; polar, cyan; glycine, gray. Interactions: hydrogen bonds to the side chain, dashed magenta line; hydrogen bonds to the main chain, solid magenta line; π–π interaction, dark green line.
