Metabolism of diafenthiuron by microsomal oxidation: procide activation and inactivation as mechanisms contributing to selectivity.
The thiourea insecticide/acaricide diafenthiuron represents a biologically inactive propesticide that requires transformation into the active carbodiimide derivative. The carbodiimide inhibits mitochondrial respiration by selective and covalent binding to the proteolipid (8 kDa) of Fo-ATPase in the inner membrane and to porin (30 kDa) in the outer membrane. The thiourea can be activated by light as well as by cytochrome P450 in the insect. To get insight into the enzymatic mechanisms of activation, model in vitro studies were performed using [14C]diafenthiuron and microsomes from various vertebrate livers and from locust Malpighian tubules. Though there was a common set of metabolites, their quantities varies significantly with the species and assay conditions. As a typical product, p-hydroxydiafenthiuron was identified in assay with rat and mouse microsomes. The sulfomonoxide predominated in hen and fish assays, whereas pig and bovine microsomes almost exclusively produced the carbodiimide. The sulfoxide was shown to be a precursor of the carbodiimide. Formation of all metabolites was dependent on the presence of NADPH and active microsomes. The effects of inhibitors and the requirement for NAPDH suggested a role of cytochrome P450-dependent monooxygenase(s) in the formation of both the hydroxylated product and the carbodiimide. FAD-dependent monooxygenases (FMOs) may also be involved in a step following sulfoxidation. These in vitro studies revealed potential mechanisms contributing to biological selectivity of the effects of a pesticide that acts in a non-selective mode at a conserved mitochondrial site.
Kayser
H
H
Syngenta Crop Protection AG, Research Biochemistry, CD-4002 Basel, Switzerland. hartmut.kayser@syngenta.com
Eilinger
P
P
D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D007300:Insect Control; D007306:Insecticides / Q000737:chemistry / Q000378:metabolism*; D008861:Microsomes / Q000201:enzymology / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism; D006899:Mixed Function Oxygenases; D010084:Oxidation-Reduction; D010670:Phenylthiourea / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism*
C071365:3-hydroxyphenylacetate 6-hydroxylase; D007306:Insecticides; C087661:diafenthiuron; D010670:Phenylthiourea; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases