Identification of a Novel Dual Inhibitor of Acetylcholinesterase and Butyrylcholinesterase: In Vitro and In Silico Studies

Raquel B M de Almeida; Deyse B Barbosa; Mayra R do Bomfim; Jéssika A O Amparo; Bruno S Andrade; Silvia L Costa; Joaquín M Campos; Jorddy N Cruz; Cleydson B R Santos; Franco H A Leite; Mariana B Botura

doi:10.3390/ph16010095

Identification of a Novel Dual Inhibitor of Acetylcholinesterase and Butyrylcholinesterase: In Vitro and In Silico Studies

Pharmaceuticals (Basel). 2023 Jan 9;16(1):95. doi: 10.3390/ph16010095.

Authors

Affiliations

¹ Department of Health, State University of Feira de Santana, Feira de Santana 44036-900, BA, Brazil.
² Institute of Health Sciences, Federal University of Bahia, Salvador 40170-110, BA, Brazil.
³ Department of Biological Sciences, State University of Southwest of Bahia, Jequié 45208-091, BA, Brazil.
⁴ Biosanitary Institute of Granada (ibs.GRANADA), University of Granada, 18071 Granada, Spain.
⁵ Department of Pharmaceutical and Organic Chemistry, Faculty of Pharmacy, Campus of Cartuja, University of Granada, 18071 Granada, Spain.
⁶ Laboratory of Modeling and Computational Chemistry, Department of Biological and Health Sciences, Federal University of Amapá, Macapá 68902-280, AP, Brazil.

Abstract

The enhancement of cholinergic functions via acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) inhibition is considered a valuable therapeutic strategy for the treatment of Alzheimer's disease. This study aimed to evaluate the in vitro effect of ZINC390718, previously filtered using computational approaches, on both cholinesterases and to characterize, using a molecular dynamics (MD) simulation, the possible binding mode of this compound inside the cholinesterase enzymes. The in vitro cytotoxicity effect was also investigated using a primary astrocyte-enriched glial cell culture. ZINC390718 presented in vitro dual inhibitory activity against AChE at a high micromolar range (IC₅₀ = 543.8 µM) and against BuChE (IC₅₀ = 241.1 µM) in a concentration-dependent manner, with greater activity against BuChE. The MD simulation revealed that ZINC390718 performed important hydrophobic and H-bond interactions with the catalytic residue sites on both targets. The residues that promoted the hydrophobic interactions and H-bonding in the AChE target were Leu67, Trp86, Phe123, Tyr124, Ser293, Phe295, and Tyr341, and on the BuChE target, they were Asp70, Tyr332, Tyr128, Ile442, Trp82, and Glu197. The cytotoxic effect of Z390718, evaluated via cell viability, showed that the molecule has low in vitro toxicity. The in vitro and in silico results indicate that ZINC390718 can be used as chemotype for the optimization and identification of new dual cholinesterase inhibitors.

Keywords: Alzheimer’s disease; acetylcholinesterase; butyrylcholinesterase; inhibitors.

Grants and funding

This research was funded by Fundação de Amparo à Pesquisa do Estado da Bahia (FAPESB, BOL0346/2017) and Programa de Pós-Graduação em Biotecnologia of Universidade Estadual de Feira de Santana (UEFS).