Inhibition of Butyrylcholinesterase with Fluorobenzylcymserine, An Experimental Alzheimer's Drug Candidate: Validation of Enzoinformatics Results by Classical and Innovative Enzyme Kinetic Analyses

Abstract

Background Selective butyrylcholinesterase (BuChE)-inhibition, increases acetylcholine (ACh) levels. In rodents, this inhibition is known to boost cognition. Also, this occurs without the typical unwanted adverse effects of acetylcholinesterase-inhibitors or AChE-Is. The novel compound, fluorobenzylcymserine (FBC), is derived from our effort to design a selective BuChE-inhibitor. Also, we wanted to check whether butyrylcholinesterase-inhibitors (BuChE-Is) possessed an edge over AChE-Is in Alzheimer’s disease (AD) in terms of efficacy and/or tolerance. Method FBC was synthesized as reported earlier while enzymatic activity of BuChE was calculated by Ellman-technique. Molecular docking was performed using Autodock4.2. We applied classical as well as innovative analyses of enzyme-kinetics for exploring “FBC:human BuChE-interaction”. The mode of inhibition and kinetic parameters were also determined. Results Docking results displayed two strong interacting sites for FBC. One of these binding sites was previously identified as a deep narrow groove having polar aromatic residues while a second site was identified during this study which displayed better interaction and was lined with aliphatic and sulphur containing residues. At low concentrations of BuChE, the IC50 was found to be very low i.e. 4.79 and 6.10 nM for 12 and 36 μg, respectively, whereas it increased exponentially by increasing the units of BuChE. Conclusion These analyses indicate that FBC is an interesting AD drug candidate that could provide a potent and partial mixed type of inhibition of human BuChE.

Molecular Interaction of Anti‐Diabetic Drugs With Acetylcholinesterase and Sodium Glucose Co‐Transporter 2

“…Most humbly, this is to state that the author, Shazi Shakil had coined the term “ENZOINFORMATICS” in 2012 and established it as a sub‐discipline of Bioinformatics [Kamal et al, ]. The computationally predicted binding interactions of CTX‐M‐15 (a bacterial enzyme) with cefotaxime (an antibiotic) by the author in 2010 [Shakil and Khan, ] perfectly matched with the X‐ray crystal of the same complex produced in the subsequent year [An et al, ]; and this is exactly the beauty of computational studies to predict a wet lab phenomenon beforehand.…”

“…Most humbly, this is to state that the author, Shazi Shakil had coined the term "ENZOINFORMATICS" in 2012 and established it as a sub-discipline of Bioinformatics [Kamal et al, 2017]. It is noteworthy to mention that this is a purely computational predictive study which does possess obvious limitations.…”

“…It is imperative that this study is meant for basic academic research purpose only and the results described herein should not be used to formulate opinion in favor or against any drug till full validated proofs involving due clinical trial protocols arrive. Most humbly, this is to state that the author, Shazi Shakil had coined the term "ENZOINFORMATICS" in 2012 and established it as a sub-discipline of Bioinformatics [Kamal et al, 2017]. The computationally predicted binding interactions of CTX-M-15 (a bacterial enzyme) with cefotaxime (an antibiotic) by the author in 2010 [Shakil and Khan, 2010] perfectly matched with the X-ray crystal of the same complex produced in the subsequent year [An et al, 2011]; and this is exactly the beauty of computational studies to predict a wet lab phenomenon beforehand.…”

Treatment of canine cognitive dysfunction with novel butyrylcholinesterase inhibitor

“…In humans with AD, a supportive co-regulating role of BChE in the brain cholinergic system was reported, and a negative role predicted elevated levels of BChE dysregulating brain AChE levels 23 . The basis for the adverse effects of approved drugs for treating patients with AD is the inhibition of cholinesterases in the basal ganglia, peripheral nervous system, and parasympathetic autonomic nervous system resulting in excess of acetylcholine 24 .…”

Molecular interaction of investigational ligands with human brain acetylcholinesterase

“…These include sotagliflozin, methotrexate, huperzine B, lycodine, dapagliflozin, and glimepiride, to name a few. Most humbly, it is worth mentioning that we had duly reviewed pertinent literature in the past and we continue with our quest of finding novel neuro drugs presently as well . Cholinergic hypothesis states that occurrence of AD might be owing to deficient production of acetylcholine, a vital neurotransmitter .…”