2-Hydroxy- N-phenylbenzamides and Their Esters Inhibit Acetylcholinesterase and Butyrylcholinesterase

Biomolecules. 2019 Nov 5;9(11):698. doi: 10.3390/biom9110698.


The development of novel inhibitors of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) represents a viable approach to alleviate Alzheimer's disease. Thirty-six halogenated 2-hydroxy-N-phenylbenzamides (salicylanilides) with various substitution patterns and their esters with phosphorus-based acids were synthesized in yields of 72% to 92% and characterized. They were evaluated for in vitro inhibition of AChE from electric eel and BuChE from equine serum using modified Ellman's spectrophotometric method. The benzamides exhibited a moderate inhibition of AChE with IC50 values in a narrow concentration range from 33.1 to 85.8 µM. IC50 values for BuChE were higher (53.5-228.4 µM). The majority of derivatives inhibit AChE more efficiently than BuChE and are comparable or superior to rivastigmine-an established cholinesterases inhibitor used in the treatment of Alzheimer's disease. Phosphorus-based esters especially improved the activity against BuChE with 5-chloro-2-{[4-(trifluoromethyl)phenyl]carbamoyl}phenyl diethyl phosphite 5c superiority (IC50 = 2.4 µM). This derivative was also the most selective inhibitor of BuChE. It caused a mixed inhibition of both cholinesterases and acted as a pseudo-irreversible inhibitor. Several structure-activity relationships were identified, e.g., favouring esters and benzamides obtained from 5-halogenosalicylic acids and polyhalogenated anilines. Both 2-hydroxy-N-phenylbenzamides and esters share convenient physicochemical properties for blood-brain-barrier penetration and thus central nervous system delivery.

Keywords: acetylcholinesterase; benzamides; butyrylcholinesterase; enzyme inhibition; esters; in vitro inhibition; phosphorus derivatives; salicylanilides.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcholinesterase / metabolism*
  • Animals
  • Benzamides / chemistry
  • Benzamides / pharmacology*
  • Butyrylcholinesterase / metabolism*
  • Cholinesterase Inhibitors / chemistry
  • Cholinesterase Inhibitors / pharmacology*
  • Electrophorus
  • Esters / chemistry
  • Esters / pharmacology*
  • Horses
  • Inhibitory Concentration 50
  • Molecular Structure
  • Phosphorus / chemistry
  • Structure-Activity Relationship


  • Benzamides
  • Cholinesterase Inhibitors
  • Esters
  • Phosphorus
  • Acetylcholinesterase
  • Butyrylcholinesterase