The endocannabinoid system (ECS) plays a key modulatory role during synaptic plasticity and homeostatic processes in the brain, and has an important role in the neurobiological processes underlying drug addiction. We have previously shown that an elevated ECS response to psychostimulant (cocaine) is involved in regulating the development and expression of cocaine-conditioned reward and cocaine sensitization. Here, we determine the involvement of ECS in alcohol addiction. We first measured the levels of endocannabinoids (eCBs) and eCB-like molecules in different regions of the brain reward system following chronic alcohol self-administration using LCMS. We found that following chronic intermittent alcohol consumption, N-oleoyl glycine (OlGly) levels were significantly elevated in the prefrontal cortex (PFC), and N-oleoyl alanine (OlAla) was significantly elevated in the PFC, nucleus accumbens (NAc) and ventral tegmental area (VTA) in a region-specific manner. We next tested whether exogenous administration of OlGly or OlAla would attenuate alcohol consumption and preference. We found that systemic administration of OlGly or OlAla (60mg/kg, intraperitoneal) during intermittent alcohol consumption significantly reduced alcohol intake and preference. These findings suggest that the ECS negatively regulates alcohol consumption and boosting selective endocannabinoids exogenously has beneficial effects against alcohol consumption and potentially in preventing relapse.