A series of deoxyvasicinone analogues were synthesized in order to produce new high efficiency deoxyvasicinone derivatives of cholinesterase (ChE) inhibitors, and their structure-activity relationships (SARs) were investigated. The compounds with six-membered rings exhibited greater ChE inhibitory activity than the compounds with other sizes of ring. C=N bonds were important for ChE inhibitory activity and selectivity for butyrylocholinoesterase (BuChE). Hydroxyl groups were able to inhibit ChE. Conversely, carbonyl groups decreased ChE inhibitory activity. Furthermore, docking studies demonstrated that proteins and ligands interacted through π-π stacking, hydrogen bonds and “water bridges”.