Kinetic parameters of OPT pesticide desulfuration by c-DNA expressed human CYPs

The role of different cytochrome P450 isoforms (CYPs) in the desulfuration of four organophosphorothionate pesticides (OPTs), namely diazinon (DIA), azinphos-methyl (AZ), chlorpyrifos (CPF) and parathion (PARA), at OPT levels representative of actual human exposure has been investigated. For this purpose c-DNA expressed human CYPs and a method, based on acetyleholinesterase (AChE) inhibition, able to detect nM levels of oxon have been used. Our results indicate that the four tested OPTs at low concentration were mainly desulfurated by CYP2B6, 2C19 and 1A2, showing K-m values in the range 0.8-5 muM and the highest efficiency (intrinsic clearance (ICL)) values. CYP3A4 was generally endowed with high K., and resulted linear up to 25-100 muM OPT, concentrations saturating the most efficient CYPs. The tentative extrapolation of the relative contribution of single CYPs, taking into account the average content of different isoforms in the human liver, indicate that CYP1A2 is the major responsible for oxon formation. Indeed this CYP catalyses the 50-90% of desulfuration reaction, depending on the OPT. As CYP3A4 activity is not completely saturated up to 100 muM OPT, and due to the high hepatic content, its contribution to oxon formation may result relevant in poisoning episodes, when individuals are exposed at high doses of OPTs. (C) 2002 Elsevier Science B.V. All rights reserved.