Fig. 1
From: High-potency ligands for DREADD imaging and activation in rodents and monkeys
New DREADD ligands displaying high in vitro DREADD affinity and potency. a Compound 13 (C13) and Compound 22 (C22) structures. b Binding competition curves of [3H]CLZ versus increasing concentrations of C13 and C22 in HEK-293 cells expressing DREADDs. C13 and C22 exhibit comparable DREADD affinity to clozapine (CLZ) with C13 showing ~twofold greater affinity than C22. CLZ and C21 competition curves from Supplementary Fig. 1 are overlaid for comparison. c, d C13 selectively blocks [3H]CLZ binding to DREADDs in mouse slices at 10 nM. Representative images of sections collected from 3 different mice are displayed and quantified in (d) as mean ± SEM. Two-way ANOVA followed by Dunnett’s test, *p < 0.05 and **p < 0.01 compared with the respective vehicle. e–h [3H]C13 binds with greater selectivity than [3H]CLZ to DREADDs in mouse and monkey brain tissue expressing AAV-hM3Dq and AAV-hM4Di, respectively. i Intraperitoneal (IP) injection of [3H]C13 readily enters the brain and accumulates in DREADDs expression areas in D1-DREADD mice. Representative images from 3 mice per condition. j–l JHU37107 (J07), JHU37152 (J52), and JHU37160 (J60) are high-affinity DREADD ligands. m Docking and molecular dynamics simulation of J60 in the ligand binding pocket of a hM4Di model. n, o J60 and J52 selectively displace [3H]CLZ at a concentration of 1 and 10 nM from hM3Dq and hM4Di expressed in mouse brain sections (n = 3 mice per condition). p, q J60 and J52 activate hM3Dq and hM4Di expressed in HEK293 cells with high potency (experiments performed 3–5 times). In all cases, data are represented as mean ± SEM. Scale bars are 1 mm. Source data are provided as a Source Data file
