Discovery of new, highly potent and selective inhibitors of BuChE - design, synthesis, in vitro and in vivo evaluation and crystallography studies

Eur J Med Chem. 2023 Mar 5:249:115135. doi: 10.1016/j.ejmech.2023.115135. Epub 2023 Jan 18.


The symptomatic and disease-modifying effects of butyrylcholinesterase (BuChE) inhibitors provide an encouraging premise for researching effective treatments for Alzheimer's disease. Here, we examined a series of compounds with a new chemical scaffold based on 3-(cyclohexylmethyl)amino-2-hydroxypropyl, and we identified a highly selective hBuChE inhibitor (29). Based on extensive in vitro and in vivo evaluations of the compound and its enantiomers, (R)-29 was identified as a promising candidate for further development. Compound (R)-29 is a potent hBuChE inhibitor (IC50 = 40 nM) with selectivity over AChE and relevant off-targets, including H1, M1, α1A and β1 receptors. The compound displays high metabolic stability on human liver microsomes (90% of the parent compound after 2 h of incubation), and its safety was confirmed through examining the cytotoxicity on the HepG2 cell line (LC50 = 2.85 μM) and hERG inhibition (less than 50% at 10 μM). While (rac)-29 lacked an effect in vivo and showed limited penetration to the CNS in pharmacokinetics studies, compound (R)-29 exhibited a procognitive effect at 15 mg/kg in the passive avoidance task in scopolamine-treated mice.

Keywords: Alzheimer's disease; Butyrylcholinesterase; Butyrylcholinesterase inhibitors; Crystallography; Passive avoidance.

MeSH terms

  • Acetylcholinesterase / metabolism
  • Alzheimer Disease* / metabolism
  • Animals
  • Butyrylcholinesterase* / metabolism
  • Cholinesterase Inhibitors / chemistry
  • Crystallography
  • Humans
  • Mice
  • Molecular Docking Simulation
  • Scopolamine / pharmacology
  • Structure-Activity Relationship


  • Butyrylcholinesterase
  • Cholinesterase Inhibitors
  • Scopolamine
  • Acetylcholinesterase