© 2023, The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.Cholinesterase enzyme family consists of acetylcholinesterase (AChE, 220.127.116.11), the major enzyme responsible for hydrolysis of acetylcholine at cholinergic synapses, and butyrylcholinesterase (BChE, 18.104.22.168) a detoxification enzyme of plasma. Statins are cholesterol-lowering medications utilized as protective medicaments in stroke and Alzheimer’s disease, which cholinesterases are associated with. Thus, in this study, we characterized the inhibitory effects and mechanisms of common statins, rosuvastatin, atorvastatin, simvastatin and lovastatin, on human erythrocyte AChE and purified serum BChE using in vitro and in silico methods. Kinetic assays identified statins as selective non-competitive inhibitors of human serum BChE. The IC50 and Km values were found as 194.7 ± 55.2 µM and 1.03 ± 0.2 µM for rosuvastatin, 492.5 ± 55.1 µM and 7.2 ± 0.3 µM for atorvastatin, 14.2 ± 0.3 µM and 202.7 ± 23.2 µM for lovastatin, and 17.6 ± 0.1 µM and 207.2 ± 13.2 µM for simvastatin, respectively. The compounds did not display considerable inhibition against AChE. Molecular docking predicted good affinity and strong interactions with the BChE active site for atorvastatin and rosuvastatin. Current study identifies rosuvastatin as the most specific and selective inhibitor of human BChE among the tested statins. As selective inhibitors of BChE statins have the potential to be re-evaluated as medicaments due to their pleiotropic effects.