Basal forebrain cholinergic neurons and oxidative stress in brain have been suggested to play an important role in the regulation of memory functions. Therefore, the present study was planned to study the effect of donepezil, an anticholinesterase antidementia drug, insulin and melatonin, an antioxidant, on memory deficit and acetylcholinesterase (AChE) activity in brain areas of scopolamine-induced amnesic mice. Memory was tested by passive avoidance (PA) test in Swiss adult male mice. A significant increase in transfer latency time (TLT) in 2nd trial as compared to 1st trial is considered as successful learning. Scopolamine (3 mg/kg i.p.) was administered 5 min prior to 1st trial to induce amnesia. AChE activity in detergent soluble (DS) and salt soluble (SS) fractions was estimated in brain areas after completion of 2nd trial. Scopolamine was effective in producing memory impairment (amnesia) which was reverted by donepezil (5 mg/kg p.o.), insulin (1 IU/kg i.p.) and melatonin (20 mg/kg p.o.). AChE activity in DS fraction of scopolamine amnesic mice was inhibited by donepezil, insulin and melatonin with varying extent in different brain regions, whereas AChE activity in SS fraction was not much affected. The results demonstrate that anti-amnesic effect of donepezil, insulin and melatonin may be mediated through enhancement of cholinergic activity.