Background and Objectives Viloxazine, which has been used to treat depression and attention-deficit/hyperactivity disorder (ADHD), has been termed a moderate-affinity, selective norepinephrine reuptake inhibitor based on high selectivity for norepinephrine relative to serotonin and dopamine transporters. However, accumulated research suggests a more complex mechanism of action, based on studies showing activity at serotonin 5-HT2C, 5-HT2B, and 5-HT7 receptors, as well as findings that viloxazine increases extracellular serotonin (along with norepinephrine and dopamine) in the rat prefrontal cortex. This in vitro pharmacology study aimed to replicate and expand prior experiments to better characterize viloxazine’s affinity for and activity at the norepinephrine transporter (NET) and individual serotonin receptors and to clarify how these effects contribute to the mechanism of action. Methods Using in vitro binding competition and functional assays and ex vivo receptor occupancy studies in rats, we assessed viloxazine activity at human NET isoforms and 5-HT2C, 5-HT2B, and 5-HT7 receptors relative to clinically relevant unbound viloxazine plasma concentrations (0.4–3.6 μM). Results Viloxazine showed moderate binding affinity for NET (inhibition constant [Ki] = 0.13 µM) and 5-HT2C (Ki = 0.66 µM), 5-HT2B (Ki = 0.83 µM), and 5-HT7 (Ki = 1.90 µM) receptors. In vitro functional studies showed viloxazine acted as a NET inhibitor, 5-HT2C partial agonist, and 5-HT2B and 5-HT7 antagonist. At clinically relevant concentrations, viloxazine could potentially occupy nearly 95% of NET, more than 80% of 5-HT2C and 5-HT2B, and 65% of 5-HT7 receptors. Subsequent ex vivo studies in rats confirmed high NET occupancy (67–94%) at clinically relevant concentrations. Conclusions These results validate previous experiments showing that viloxazine, in addition to displaying high NET occupancy, acts as a partial agonist at 5-HT2C receptors and an antagonist at 5-HT2B and 5-HT7 receptors at clinically relevant concentrations for ADHD treatment. Therefore, both NET inhibition and serotonin receptor activity may contribute to viloxazine’s clinical efficacy. These findings are contributing to a renewed understanding of viloxazine’s pharmacodynamic profile and likely multimodal mechanism of action.

Updated Viloxazine Pharmacology: Experiments Establish Norepinephrine Transporter Occupancy and Serotonin 5-HT_2C, 5-HT_2B, and 5-HT₇ Receptor Binding at Therapeutically Relevant Concentrations

期刊论文
出版模式：

开放获取
发表日期：

2026年4月3日

查看全文

Jennie Garcia-Olivares,
Brittney Yegla,
Jennifer Koch,
Chungping Yu,
Jonathan Rubin

摘要

Background and Objectives

Viloxazine, which has been used to treat depression and attention-deficit/hyperactivity disorder (ADHD), has been termed a moderate-affinity, selective norepinephrine reuptake inhibitor based on high selectivity for norepinephrine relative to serotonin and dopamine transporters. However, accumulated research suggests a more complex mechanism of action, based on studies showing activity at serotonin 5-HT_2C, 5-HT_2B, and 5-HT₇ receptors, as well as findings that viloxazine increases extracellular serotonin (along with norepinephrine and dopamine) in the rat prefrontal cortex. This in vitro pharmacology study aimed to replicate and expand prior experiments to better characterize viloxazine’s affinity for and activity at the norepinephrine transporter (NET) and individual serotonin receptors and to clarify how these effects contribute to the mechanism of action.

Methods

Using in vitro binding competition and functional assays and ex vivo receptor occupancy studies in rats, we assessed viloxazine activity at human NET isoforms and 5-HT_2C, 5-HT_2B, and 5-HT₇ receptors relative to clinically relevant unbound viloxazine plasma concentrations (0.4–3.6 μM).

Results

Viloxazine showed moderate binding affinity for NET (inhibition constant [K_i] = 0.13 µM) and 5-HT_2C (K_i = 0.66 µM), 5-HT_2B (K_i = 0.83 µM), and 5-HT₇ (K_i = 1.90 µM) receptors. In vitro functional studies showed viloxazine acted as a NET inhibitor, 5-HT_2C partial agonist, and 5-HT_2B and 5-HT₇ antagonist. At clinically relevant concentrations, viloxazine could potentially occupy nearly 95% of NET, more than 80% of 5-HT_2C and 5-HT_2B, and 65% of 5-HT₇ receptors. Subsequent ex vivo studies in rats confirmed high NET occupancy (67–94%) at clinically relevant concentrations.

Conclusions

These results validate previous experiments showing that viloxazine, in addition to displaying high NET occupancy, acts as a partial agonist at 5-HT_2C receptors and an antagonist at 5-HT_2B and 5-HT₇ receptors at clinically relevant concentrations for ADHD treatment. Therefore, both NET inhibition and serotonin receptor activity may contribute to viloxazine’s clinical efficacy. These findings are contributing to a renewed understanding of viloxazine’s pharmacodynamic profile and likely multimodal mechanism of action.

Updated Viloxazine Pharmacology: Experiments Establish Norepinephrine Transporter Occupancy and Serotonin 5-HT2C, 5-HT2B, and 5-HT7 Receptor Binding at Therapeutically Relevant Concentrations

摘要

Updated Viloxazine Pharmacology: Experiments Establish Norepinephrine Transporter Occupancy and Serotonin 5-HT_2C, 5-HT_2B, and 5-HT₇ Receptor Binding at Therapeutically Relevant Concentrations