Pharmacotherapy for Alcohol Use Disorder (AUD) may be improved by targeting mechanisms by which alcohol interferes with dopamine (DA) pathways in the reward circuitry. Alcohol acts on inhibitory glycine receptors (GlyR) in the nucleus Accumbens (nAc) and treatment with GlyR agonists raise basal DA, attenuate alcohol-induced DA release in the nAc and reduce alcohol intake in rats. Combined treatment with varenicline+bupropion produces additive effects on nAc DA output and abolishes the alcohol deprivation effect (ADE) in rats, predictive of clinical efficacy. This study examines whether the glycine transporter-1(GlyT1)-inhibitor Org24598, an indirect GlyR agonist, enhances the ADE-reducing and DA elevating action of varenicline+bupropion.Effects on alcohol intake, the ADE and nAc glycine and DA levels were examined following systemic treatment with threshold doses for Org24598, bupropion, and varenicline in monotherapy or combined, using a free-choice alcohol consumption model with an ensuing ADE paradigm, and in vivo microdialysis in male Wistar rats.All treatment regimens abolished the ADE but only the effect produced by the triple combination was statistically significant compared to vehicle. Hence, addition of Org24598 may enhance the ADE-reducing action of varenicline+bupropion. Org24598 raised nAc glycine but not DA output in monotherapy. Varenicline+bupropion produced a substantial elevation in nAc DA output that was enhanced following addition of Org24598.Conceivably, the blockade of the ADE is achieved by the triple combination enhancing nAc DA transmission in complementary ways, thereby alleviating hypodopaminergia and negative reinforcement to drink. Ultimately, combining GlyR agonists with varenicline+bupropion may constitute a new pharmacological treatment principle for AUD.