Inhibition of brain carnitinepalmitoyl-transferase-1 (CPT-1) is reported to decrease food intake and body weight. Yet, the fatty acid synthase (FAS) inhibitor and CPT-1 stimulator C75 produces hypophagia and weight loss when given to rodents intracerebroventricularly (i.c.v.). Thus, roles and contributions of altered brain CPT-1 activity and fatty acid oxidation in these phenomena remain unclarified. We administered compounds targeting FAS or CPT-1 to mice by single i.c.v. bolus, and examined acute and prolonged effects on feeding and body weight. C75 decreased food intake rapidly and potently at all doses (1-56 mol), and dose dependently inhibited intake on day 1. Dose dependent weight loss on day 1 persisted through four days of post-injection monitoring. The FAS inhibitor cerulenin (560 mol) produced hypophagia for one day, weight loss for two days, and weight regain to vehicle control by day 3. The CPT-1 inhibitor etomoxir (32, 320 mol) did not alter overall day 1 feeding. However, etomoxir attenuated the hypophagia produced by C75, indicating that CPT-1 stimulation is important for C75’s effect. A novel compound, C89b, was characterized in vitro as a selective inhibitor of CPT-1 that does not affect fatty acid synthesis. C89b (100, 320 mol) decreased feeding in mice for three days, with persistent weight loss for six days, without producing conditioned taste aversion. Similarly, intraperitoneal administration decreased feeding and body weight without producing conditioned taste aversion. These results suggest a role for brain CPT-1 in the regulation of energy balance and implicate CPT-1 stimulation as a pharmacological approach to weight loss.