In Silico and In Vitro Evaluation of Calm Supplement Constituents on Antidepressant-Related Molecular Targets and Neuroplasticity-Associated Gene Expression

Background: Depression-related mechanisms involve not only monoaminergic neurotransmission but also neurotrophic signaling, synaptic plasticity and intracellular survival pathways. Materials and Methods: In this study, the potential neuropharmacological properties of Calm, a multi-component supplement product, were evaluated using in silico and in vitro approaches. Molecular docking analyses were performed to estimate the binding profiles of Calm-derived constituents and reference antidepressants, sertraline and fluoxetine, against SLC6A4, MAOA, AKT1 and NTRK2. In parallel, SH-SY5Y human neuroblastoma cells were treated with Calm, and MTT-based metabolic activity and the relative expression levels of BDNF, NTRK2 and SYN1 were assessed after 24 h. Results: Docking results showed that several Calm constituents, particularly apigenin, rosmarinic acid, xanthohumol and valerenic acid, exhibited favorable predicted interactions with SLC6A4 and NTRK2. Calm treatment increased MTT signal at low concentrations, with significant increases observed at 10, 30, and 60 µg/mL. However, 24 h exposure to 10 µg/mL Calm significantly reduced BDNF, NTRK2 and SYN1 expression levels. Conclusions: These findings suggest that Calm constituents may interact with antidepressant-related molecular targets and modulate neuroplasticity-associated transcriptional markers. However, the results should be considered preliminary and require protein-level and functional validation.