title: Esterase metabolism of cholinesterase inhibitors using rat liver in vitro.
abstract: A variety of chemicals, such as organophosphate (OP) and carbamate pesticides, nerve agents, and industrial chemicals, inhibit acetylcholinesterase (AChE) leading to overstimulation of the cholinergic nervous system. The resultant neurotoxicity is similar across mammalian species; however, the relative potencies of the chemicals across and within species depend in part on chemical-specific metabolic and detoxification processes. Carboxylesterases and A-esterases (paraoxonases, PON) are two enzymatic detoxification pathways that have been widely studied. We used an in vitro system to measure esterase-dependent detoxification of 15 AChE inhibitors. The target enzyme AChE served as a bioassay of inhibitor concentration following incubation with detoxifying tissue. Concentration-inhibition curves were determined for the inhibitor in the presence of buffer (no liver), rat liver plus calcium (to stimulate PONs and thereby measure both PON and carboxylesterase), and rat liver plus EGTA (to inhibit calcium-dependent PONs, measuring carboxylesterase activity). Point estimates (concentrations calculated to produce 20, 50, and 80% inhibition) were compared across conditions and served as a measure of esterase-mediated detoxification. Results with well-known inhibitors (chlorpyrifos oxon, paraoxon, methyl paraoxon, malaoxon) were in agreement with the literature, serving to support the use of this assay. Only a few other inhibitors showed slight or a trend towards detoxification via carboxylesterases or PONs (mevinphos, aldicarb, oxamyl). There was no apparent PON- or carboxylesterase-mediated detoxification of the remaining inhibitors (carbofuran, chlorfenvinphos, dicrotophos, fenamiphos, methamidophos, methomyl, monocrotophos, phosphamidon), suggesting that the influence of esterases on these chemicals is minimal. Thus, generalizations regarding these metabolic pathways may not be appropriate. As with other aspects of AChE inhibitors, their metabolic patterns appear to be chemical-specific.
authors.affiliation: Toxicity Assessment Division (MD B105-04), National Health and Environmental Effects Research Laboratory, Office of Research and Development, US Environmental Protection Agency, Research Triangle Park, NC 27711, USA. Moser.ginger@epa.gov
authors.forename: V C
authors.initials: VC
authors.lastname: Moser
authors.forename: S
authors.initials: S
authors.lastname: Padilla
mesh_terms: D000818:Animals; D043303:Aryldialkylphosphatase / Q000187:drug effects / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D043182:Carboxylesterase / Q000187:drug effects / Q000378:metabolism; D004390:Chlorpyrifos / Q000031:analogs & derivatives / Q000493:pharmacokinetics / Q000494:pharmacology; D002800:Cholinesterase Inhibitors / Q000493:pharmacokinetics* / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004950:Esterases / Q000187:drug effects / Q000378:metabolism*; D066298:In Vitro Techniques; D008658:Inactivation, Metabolic; D008099:Liver / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D008297:Male; D051381:Rats; D020318:Rats, Long-Evans
chemical_list: D002800:Cholinesterase Inhibitors; C009618:O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphate; D004950:Esterases; D043182:Carboxylesterase; D043303:Aryldialkylphosphatase; D004390:Chlorpyrifos; D002118:Calcium