Cysteine-Targeted Insecticides against A. gambiae Acetylcholinesterase Are Neither Selective nor Reversible Inhibitors

Abstract

Acetylcholinesterase cysteine-targeted insecticides against malaria vector Anopheles gambia and other mosquitos have already been introduced. We have applied the olefin metathesis for the preparation of cysteine-targeted insecticides in high yields. The prepared compounds with either a succinimide or maleimide moiety were evaluated on Anopheles gambiae and human acetylcholinesterase with relatively high irreversible inhibition of both enzymes but poor selectivity. The concept of cysteine binding was not proved by several methods, and poor stability was observed of the chosen most potent/selective compounds in a water/ buffer environment. Thus, our findings do not support the proposed concept of cysteine-targeted selective insecticides for the prepared series of succinimide or maleimide compounds.

Synthesis,in vitroscreening and molecular docking of isoquinolinium-5-carbaldoximes as acetylcholinesterase and butyrylcholinesterase reactivators

“…The recombinant form of human AChE/BChE was prepared in Department of Chemistry (Faculty of Science, University of Hradec Kralove, Hradec Kralove, Czech Republic) 12 and purified using NGC Medium-Pressure Chromatography System (Bio-Rad, Hercules, CA). The total volume of 6-8 ml of medium containing secreted protein was desalted using 5 ml HiTrap Desalting column (GE Healthcare, Chicago, IL) equilibrated with buffer A (20 mM sodium phosphate buffer, 150 mM NaCl, 15 mM imidazole and 20% glycerol; pH 7.4).…”

“…The reactivators as pralidoxime (9), asoxime (syn. HI-6; 10), obidoxime (11), trimedoxime (12), and methoxime (13) (Figure 2) are approved as an antidotes against nerve agents, but unfortunately, their efficacy in suppressing acute toxic effects of all types of nerve agents is rather limited 10 . However, these are the most studied and commercially available reactivators 8 .…”

Pyridinium-2-carbaldoximes with quinolinium carboxamide moiety are simultaneous reactivators of acetylcholinesterase and butyrylcholinesterase inhibited by nerve agent surrogates

“…The hr AChE and hr BChE were purified using NGC Medium-Pressure Chromatography System (Bio-Rad, Hercules, CA) 18 . The total volume of 6–8 mL of medium containing secreted protein was desalted using 5 mL HiTrap Desalting column (GE Healthcare, Chicago, IL) equilibrated with buffer A (20 mM sodium phosphate buffer, 150 mM NaCl, 15 mM imidazole, and 20% glycerol; pH 7.4).…”

1,2,4-Triazole-based anticonvulsant agents with additional ROS scavenging activity are effective in a model of pharmacoresistant epilepsy

“…The reaction mixture contained 10 mL of hrAChE (70 ng/mL protein final concentration) or hrBChE (220 ng/mL protein final concentration), 10 mL of tested compound (1, 10, 100, and 500 mM final concentration), 20 mL of 5,5 0 -dithiobis-2-nitrobenzoic acid (DTNB; 500 mM final concentration), and 60 mL of 20 mM Na-phosphate buffer (pH 7.4). The mixture was pre-incubated at 37 C for 15 min and subsequently 10 mL substrate acetylthiocholine iodide (ATChI) or butyrylthiochiline iodide (BTChI) was added to the final concentration of 1000 mM 24 . The final volume of reaction mixture was 100 mL.…”