Combination of human acetylcholinesterase and serum albumin sensing surfaces as highly informative analytical tool for inhibitor screening

• Published in: Journal of pharmaceutical and biomedical analysis 2018-06, Vol.155, p.177-184
• Main Authors: Fabini, Edoardo, Tramarin, Anna, Bartolini, Manuela
• Format: Article
• Language: English
• Subjects: Acetylcholinesterase inhibitors; Human recombinant acetylcholinesterase; Human serum albumin; Kinetic rate constants; Residence time; Surface plasmon resonance
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2018.03.060

[Display omitted] •An SPR-based assay to monitor inhibitor–hAChE interactions was developed.•A functional human recombinant AChE-sensing surface was obtained.•Steady-state affinity for donepezil, galantamine, tacrine, edrophonium was measured.•Kinetic rate constants for all drug–hAChE interactions were determined.•Drug–AChE residence time and HSA binding levels were quantified. In the continuous research for potential drug lead candidates, the availability of highly informative screening methodologies may constitute a decisive element in the selection of best-in-class compounds. In the present study, a surface plasmon resonance (SPR)-based assay was developed and employed to investigate interactions between human recombinant AChE (hAChE) and four known ligands: galantamine, tacrine, donepezil and edrophonium. To this aim, a sensor chip was functionalized with hAChE using mild immobilization conditions to best preserve enzyme integrity. Binding affinities and, for the first time, kinetic rate constants for all drug–hAChE complexes formation/disruption were determined. Inhibitors were classified in two groups: slow-reversible and fast-reversible binders according to respective target residence time. Combining data obtained on drug–target residence time with data obtained on serum albumin binding levels, a good correlation with potency, plasma protein binding in vivo, and administration regimen was found. The outcomes of this work demonstrated that the developed SPR-based assay is suitable for the screening, the binding affinity ranking and the kinetic evaluation of hAChE inhibitors. The method proposed ensures a simpler and cost-effective assay to quantify kinetic rate constants for inhibitor–hAChE interaction as compared with other proposed and published methods. Eventually, the determination of residence time in combination with preliminary ADME studies might constitute a better tool to predict in vivo behaviour, a key information for the research of new potential drug candidates.