Combined effect of organophosphorus hydrolase and oxime on the reactivation rate of diethylphosphoryl-acetylcholinesterase conjugates.
Reactivation of inhibited acetylcholinesterase (AChE) is essential for rapid recovery after organophosphate (OP) poisoning. However, following administration of an oxime reactivator, such as pralidoxime mesylate (P2S), in patients poisoned with certain diethylphosphorothioate pesticides, no reactivation is observed, presumably due to reinhibition by circulating anti-cholinesterase OPs. Pretreatment alone with organophosphorus hydrolases (OPH) that are capable of rapidly hydrolyzing OPs was demonstrated, in animals, to confer significant protection against OP toxicity. One strategy to augment the potentially therapeutic scope of OPHs is a combined post-exposure treatment consisting of a drug(s) commonly used against OP toxicity and a suitable hydrolase. In this study, we examined the in vitro ability of OPH from Pseudomonas sp. (OPHps) to prevent reinhibition of P2S-reactivated AChE by excess OPs. The kinetic parameters of the reactivation of a series of diethylphosphoryl-AChE (DEP--AChE) conjugates, obtained by the use of various diethylphosphates, were determined and compared with the rates of reactivation in the presence of OPHps, with and without the OP inhibitors in the reactivation medium. Extrapolation of the in vitro results to in vivo conditions suggests that an OPHps concentration as low as 1 microgram/mL blood would result in a 100-fold decrease in the concentration of circulating anti-AChE pesticides within less than one blood-circulation time, thereby minimizing reinhibition of the reactivated enzyme. Thus, for DEP-based pesticides, the combination of P2S-OPH treatment can significantly improve clinical recovery after OP intoxication. In addition, it is shown here for the first time that an OPH can effectively hydrolyze quaternary ammonium-containing OPs. This indicates that hydrolysis of phosphorylated oximes, toxic side products of oxime treatment, may also be accelerated by OPHs.
Ashani
Y
Y
Israel Institute for Biological Research, Ness-Ziona, Israel. yashani@netvision.net.il
Leader
H
H
Rothschild
N
N
Dosoretz
C
C
D000110:Acetylcholinesterase / Q000378:metabolism*; D000818:Animals; D043303:Aryldialkylphosphatase; D002800:Cholinesterase Inhibitors / Q000493:pharmacokinetics* / Q000506:poisoning; D002801:Cholinesterase Reactivators / Q000494:pharmacology*; D004950:Esterases / Q000378:metabolism*; D008658:Inactivation, Metabolic; D007306:Insecticides / Q000493:pharmacokinetics* / Q000506:poisoning; D007700:Kinetics; D009943:Organophosphorus Compounds; D010091:Oximes / Q000494:pharmacology*; D011485:Protein Binding; D011549:Pseudomonas / Q000201:enzymology; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity
D002800:Cholinesterase Inhibitors; D002801:Cholinesterase Reactivators; D007306:Insecticides; D009943:Organophosphorus Compounds; D010091:Oximes; D011994:Recombinant Proteins; D004950:Esterases; D000110:Acetylcholinesterase; D043303:Aryldialkylphosphatase