Secondary metabolites from natural products are potential sources of acetylcholinesterase inhibitors, which are key enzymes in the treatment of many neurodegenerative diseases. Inspired by the reported activities of alkaloids herein we report the chemical investigation on the chemical constituents from Beilschmiedia insignis and their acetylcholinesterase inhibitory activity. Isolation of the stem bark of B. insignis led to the isolation and identification of five aporphine alkaloids, namely isocorydine (1), norisocorydine (2), (+)-laurotetanine (3), (+)N-methyllaurotetanine (4), and (+)-boldine (5), together with beta-sitosterol (6), beta-sitostenone (7), lupeol (8), and lupenone (9). The chemical structures of these compounds were obtained by analysis of their spectroscopic data, as well as the comparison with that of reported data. Acetylcholinesterase inhibitory activity revealed that all isolated alkaloids were found to inhibit AChE with percentage inhibition values ranged from 44.9 to 74.5%. This is the first report on phytochemicals from B. insignis.