Fig. 1

Identification of convolamine as an S1R positive modulator. (a) In vivo phenotypic drug screening in wfs1abKO zebrafish showing the effect of repositionable drugs on locomotor response in wfs1abWT controls (Loc WT %), on hyperlocomotor response in wfs1abKO zebrafish (Hyperloc KO %) and calculated PI. Convolamine is identified as MED0092. (b) Two-way graphic representation of the screening data in (a). Dots in green correspond to 0.8 < PI <1.2 and dots in yellow to 0.5 < PI <1.5. Only convolamine located within the green circle corresponding to effects on Loc WT % and Hyperloc KO % limited to 20%. (c) Dose–response effect of convolamine on inhibition of specific 3H-radioligand binding to S1R and S2R binding sites. Estimated IC50 are indicated in the graph. (d) The in vitro pharmacological screening on 47 receptors and binding sites showed only marked inhibition of convolamine on CB2 receptors, Kv channels, 5-HT3 receptor binding and stimulation of 5-HT1B receptors, at 0.1 or 1 μM. (e) S1R/BiP dissociation assay for PRE-084 (estimated IC50 of 468 nM), NE-100 and convolamine. GFP-S1R-oe CHO cells were incubated with the test compound for 30 min at 37°C before GFP immunoprecipitation. (f) Impact of fixed concentrations of convolamine (0.1–10 μM) on PRE-084-induced dissociation of S1R from BiP. Insert: the shift in IC50 values for PRE-084 was plotted versus Convolamine concentration and showed an IC50 for the positive modulatory effect of Convolamine at 289 nM