This study investigates the modulation of anxiety through the blockade of anandamide hydrolysis, an enzyme responsible for the degradation of the endogenous cannabinoid anandamide. The researchers developed a new class of inhibitors, URB532 and URB597, which selectively block fatty acid amide hydrolase (FAAH) without affecting other serine hydrolases. These inhibitors were found to enhance brain levels of anandamide and exhibit benzodiazepine-like properties in rats, reducing anxiety-like behaviors in the elevated zero-maze test and suppressing isolation-induced vocalizations. The effects were prevented by CB1 receptor blockade, suggesting that anandamide plays a role in emotional regulation. The study highlights the potential of FAAH inhibition as a novel approach to anti-anxiety therapy, offering a mechanism that avoids the unwanted side effects of direct cannabinoid agonists.This study investigates the modulation of anxiety through the blockade of anandamide hydrolysis, an enzyme responsible for the degradation of the endogenous cannabinoid anandamide. The researchers developed a new class of inhibitors, URB532 and URB597, which selectively block fatty acid amide hydrolase (FAAH) without affecting other serine hydrolases. These inhibitors were found to enhance brain levels of anandamide and exhibit benzodiazepine-like properties in rats, reducing anxiety-like behaviors in the elevated zero-maze test and suppressing isolation-induced vocalizations. The effects were prevented by CB1 receptor blockade, suggesting that anandamide plays a role in emotional regulation. The study highlights the potential of FAAH inhibition as a novel approach to anti-anxiety therapy, offering a mechanism that avoids the unwanted side effects of direct cannabinoid agonists.