Rapid Mechanistic Evaluation and Parameter Estimation of Putative Inhibitors in a Single-Step Progress-Curve Analysis: The Case of Horse Butyrylcholinesterase

Abstract

Highly efficient and rapid lead compound evaluation for estimation of inhibition parameters and type of inhibition is proposed. This is based on a single progress-curve measurement in the presence of each candidate compound, followed by the simultaneous analysis of all of these curves using the ENZO enzyme kinetics suite, which can be implemented as a web application. In the first step, all of the candidate ligands are tested as competitive inhibitors. Where the theoretical curves do not correspond to the experimental data, minimal additional measurements are added, with subsequent processing according to modified reaction mechanisms.

New Sesquiterpenoids from the Fermented Broth of Termitomyces albuminosus and Their Anti-Acetylcholinesterase Activity

“…As mentioned above, sesquiterpenoids with anti-acetylcholinesterase activity could be a potential natural therapeutic agent for Alzheimer’s disease. However, the inhibition mechanistic and action model of the above inhibitors, which were screened by the limited methods, were unclear [25]. More data including the dissociation constant and kinetics parameters are needed for unveiling their reaction mechanism [26].…”

The Removal of Time–Concentration Data Points from Progress Curves Improves the Determination of Km: The Example of Paraoxonase 1

“…To test the performance of iFIT on potentially troublesome curves for reactions which include inhibition by product, we compared the different programs on two additional sets of progress curves for the enzymes: (1) penicillin amidase, cleaving the artificial substrate NIPAB, with data that had already been published by Zavrel et al [10] and reanalyzed by Goličnik [22], and (2) butyrylcholinesterase, cleaving the artificial substrate butyryltiocholine, with our own set of measurements, based on methods used by Stojan [23]. Both of these results are presented fully in the Supplementary Material.…”

Development of potent reversible selective inhibitors of butyrylcholinesterase as fluorescent probes

“…The progress-curve analysis was performed using the ENZO web application, implemented at www.enzo.cmm.ki.si [20][21][22] . This programme is designed to generate differential equations from drawn reaction schemes and subsequently to fit the coefficients of these equations according to least squares methodology to reproduce the experimental data using a numerical integration algorithm.…”