Identification of acetylcholinesterase inhibitors using homogenous cell‐based assays in quantitative high‐throughput screening platforms. Issue 5 (6th April 2017)

• Record Type: Journal Article
• Authors: Li, Shuaizhang; Huang, Ruili; Solomon, Samuel; Liu, Yitong; Zhao, Bin; Santillo, Michael F.; Xia, Menghang
• Is Part Of: Biotechnology journal. Volume 12:Issue 5(2017)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2017-04-06
• Subjects: Acetylcholinesterase (AChE) -- AChE inhibitors -- Cell‐based AChE assay -- Quantitative high‐throughput screening (qHTS); Biotechnology -- Periodicals; 660.605
• DOI: 10.1002/biot.201600715
• Languages: English
• ISSNs: 1860-6768

Abstract

Abstract: Acetylcholinesterase (AChE) is an enzyme responsible for metabolism of acetylcholine, a neurotransmitter associated with muscle movement, cognition, and other neurobiological processes. Inhibition of AChE activity can serve as a therapeutic mechanism, but also cause adverse health effects and neurotoxicity. In order to efficiently identify AChE inhibitors from large compound libraries, homogenous cell‐based assays in high‐throughput screening platforms are needed. In this study, a fluorescent method using Amplex Red (10‐acetyl‐3, 7‐dihydroxyphenoxazine) and the Ellman absorbance method were both developed in a homogenous format using a human neuroblastoma cell line (SH‐SY5Y). An enzyme‐based assay using Amplex Red was also optimized and used to confirm the potential inhibitors. These three assays were used to screen 1368 compounds, which included a library of pharmacologically active compounds (LOPAC) and 88 additional compounds from the Tox21 program, at multiple concentrations in a quantitative high‐throughput screening (qHTS) format. All three assays exhibited exceptional performance characteristics including assay signal quality, precision, and reproducibility. A group of inhibitors were identified from this study, including known (e.g. physostigmine and neostigmine bromide) and potential novel AChE inhibitors (e.g. chelerythrine chloride and cilostazol). These results demonstrate that this platform is a promising means to profile large numbers of chemicals that inhibit AChE activity. Abstract : Compounds that inhibit acetylcholinesterase activity have both pharmacological and toxicological significance. Three high‐throughput assays were developed and validated to identify acetylcholinesterase inhibitors by screening a collection of 1368 compounds. All three assays exhibited exceptional performance and identified both known and novel inhibitors, which shows that these methods can obtain important pharmacological and toxicological information from large compound libraries.