Centanafadine is a dual inhibitor of norepinephrine (NE)/dopamine (DA) transporter, also inhibits serotonin transporter (IC50s: 6 nM, 38 nM, and 83 nM for human NE, DA, and serotonin transporter).

NE (human):6 nM , 5-HT (human):83 nM , DA (human):38 nM

Centanafadine (EB-1020) preferentially inhibits monoamine reuptake in cloned cell lines with human transporters, showing high affinity for NE (IC50: 6 nM) and DA (IC50: 38 nM) transporters, while exhibiting a lesser effect on the 5-HT transporter (IC50: 83 nM).

In microdialysis studies, Centanafadine significantly increases NE and DA levels in the rat prefrontal cortex in vivo, with peak increases of 375% and 300%, respectively, primarily affecting NE, and elevates DA extracellular concentrations in the striatum to 400% of baseline. Centanafadine dose-dependently reduces immobility in the mouse tail suspension test of depression to 13% of control levels, without stimulating locomotor activity in adult rats at optimal doses, and inhibits locomotor hyperactivity in juvenile rats lesioned with the neurotoxin 6-hydroxydopamine (100 μg intracisternal) as neonates.