TY - JOUR T1 - Inhibition and Activation of the Human Liver Microsomal and Human Cytochrome P450 3A4 Metabolism of Testosterone by Deployment-Related Chemicals JF - Drug Metabolism and Disposition JO - Drug Metab Dispos SP - 384 LP - 391 DO - 10.1124/dmd.31.4.384 VL - 31 IS - 4 AU - Khawja A. Usmani AU - Randy L. Rose AU - Ernest Hodgson Y1 - 2003/04/01 UR - http://dmd.aspetjournals.org/content/31/4/384.abstract N2 - Cytochrome P450 (P450) enzymes are major catalysts involved in the metabolism of xenobiotics and endogenous substrates such as testosterone (TST). Major TST metabolites formed by human liver microsomes include 6β-hydroxytestosterone (6β-OHTST), 2β-hydroxytestosterone (2β-OHTST), and 15β-hydroxytestosterone (15β-OHTST). A screen of 16 cDNA-expressed human P450 isoforms demonstrated that 94% of all TST metabolites are produced by members of the CYP3A subfamily with 6β-OHTST accounting for 86% of all TST metabolites. Similar Km values were observed for production of 6β-, 2β-, and 15β-OHTST with human liver microsomes (HLM) and CYP3A4. However, Vmaxand CLint were significantly higher for 6β-OHTST than 2β-OHTST (∼18-fold) and 15β-OHTST (∼40-fold). Preincubation of HLM with a variety of ligands, including chemicals used in military deployments, resulted in varying levels of inhibition or activation of TST metabolism. The greatest inhibition of TST metabolism in HLM was following preincubation with organophosphorus compounds, including chlorpyrifos, phorate, and fonofos, with up to 80% inhibition noticed for several metabolites including 6β-OHTST. Preincubation of CYP3A4 with chlorpyrifos, but not chlorpyrifos-oxon, resulted in 98% inhibition of TST metabolism. Phorate and fonofos also inhibited the production of most primary metabolites of CYP3A4. Kinetic analysis indicated that chlorpyrifos was one of the most potent inhibitors of major TST metabolites followed by fonofos and phorate. Chlorpyrifos, fonofos, and phorate inhibited major TST metabolites noncompetitively and irreversibly. Conversely, preincubation of CYP3A4 with pyridostigmine bromide increased metabolite levels of 6β-OHTST and 2β-OHTST. Preincubation of human aromatase (CYP19) with the test chemicals had no effect on the production of the endogenous estrogen, 17β-estradiol. The American Society for Pharmacology and Experimental Therapeutics ER -