Effects of the Neutral CB1 Receptor Antagonist AM6527 on Spontaneous, Consummatory, and Motivated Behavior in Mice

Abstract

RationaleThe cannabinoid type-1 receptor (CB1R) signaling pathway plays a central role in regulating motivational and feeding behaviors. Neutral CB1R antagonists represent a promising therapeutic class with potentially fewer adverse effects than inverse agonists, yet their behavioral effects remain incompletely characterized. ObjectivesWe investigated the behavioral profile of AM6527, orally bioavailable neutral CB1R antagonist, across naturalistic and operant paradigms in male mice. To evaluate dopaminergic involvement in AM6527s effects, we employed several pharmacological interventions. ResultsUsing machine learning-based Motion-Sequencing (MoSeq), which parses spontaneous behavior into sub-second syllables, we found that AM6527 did not affect overall speed in an open field, however, it increased the self-directed behaviors and reduced specific locomotor syllables at the highest dose tested. In a naturalistic reward consumption paradigm, AM6527 produced a dose-dependent reduction in milk intake. Operant conditioning paradigms revealed robust suppression of motivated responding on fixed ratio-3 and progressive ratio (PR) schedules for palatable milk reward, with the greatest impact on high-baseline performers under PR conditions. To understand the dopaminergic involvement, we co-administered dopaminergic drugs (targeting D1R, D2R, or dopamine transporter) which resulted in partial rescue of operant responding, indicating dopaminergic and non-dopaminergic contributions to AM6527s observed behavioral effects. ConclusionOur findings suggest that neutral CB1R antagonism suppresses consummatory and motivated behaviors via dopamine-dependent and -independent mechanisms. By leveraging sub-second behavioral analysis with MoSeq, we further reveal distinct changes in spontaneous behavior, underscoring the relevance of CB-based treatments for maladaptive appetitive and motivational states in both psychiatric and metabolic disorder.