Discovery of Multi-Functional Lead Compounds Originating from Traditional Chinese Medicine for Developing Anti-Depressive Agents via Virtual Screening

The increasing prevalence of depression has become a global health issue. Currently approved anti-depressive including 5-hydroxytryptamine (5-HT), dopamine (DA), norepinephrine (NE), triple reuptake inhibitors (TRIs) and glutamate N-methyl-D-aspartate (NMDA) receptor antagonists have limited effects because of their insufficient efficacy and/or slow onset of action. Developing multifunctional antidepressants that can modulate 5-HT, DA, NE, and NMDA simultaneously can potentially overcome the current drug defects. This study aimed to explore leads for the development of multi-functional anti-depressive agents that simultaneous triple reuptake inhibitory and NMDA-GluN2B receptor antagonistic activities This study aimed to explore leads for the development of multi-functional anti-depressive agents that simultaneous triple reuptake inhibitory and NMDA-GluN2B receptor antagonistic activities. Potential leads were screened virtually from the TCMSP database based on the 3D-Pharmacophore model of TRIs followed by the molecular docking into NMDA-GluN2B receptor, BBB score, and the in-silico toxicity evaluation. The biological activities of discovered leads on 5-HT, NE, and DA reuptake and their effect on the NMDA-GluN2B receptor were evaluated via radio-labeled neurotransmitters and competition radio-ligand binding experiment with [3H] ifenprodil, respectively. Lastly, the antidepressant effect of these potential leads was determined in vivo through the forced swim test in mice Two compounds were attained as potential leads after the aforementioned experiments. Further in vitro biological evaluation identified Hit-2 as a promising lead that exerted favorable triple 5-HT/DA/NE reuptake inhibitory activity (66.98% inhibition rate at 10μM against hNET, 73.01% inhibition rate at 1μM against hDAT and 86.27% inhibition rate at 1μM against hSERT), as well as potent NMDA-GluN2B receptor antagonistic activity (Ki=115.73±3.54nM). The antidepressant activity of Hit-2 was confirmed through in vivo experiments Hit-2 not only simultaneously inhibited the reuptake of 5-HT, DA, and NE, and acted as an NMDA-GluN2B receptor antagonist in vitro but also showed in vivo antidepressant activity. These findings may serve as a structural basis for the further development of multi-functional anti-depressive agents. none