title: Metabolic activation of pradefovir by CYP3A4 and its potential as an inhibitor or inducer.
abstract: Metabolic activation of pradefovir to 9-(2-phosphonylmethoxyethyl)adenine (PMEA) was evaluated by using cDNA-expressed CYP isozymes in portal vein-cannulated rats following oral administration and in human liver microsomes. The enzyme induction potential of pradefovir was evaluated in rats following multiple oral dosing and in primary cultures of human hepatocytes. The results indicated that CYP3A4 is the only cDNA-expressed CYP isozyme catalyzing the conversion of pradefovir to PMEA. Pradefovir was converted to PMEA in human liver microsomes with a K(m) of 60 microM, a maximum rate of metabolism of 228 pmol/min/mg protein, and an intrinsic clearance of about 359 ml/min. Addition of ketoconazole and monoclonal antibody 3A4 significantly inhibits the conversion of pradefovir to PMEA in human liver microsomes, suggesting the predominant role of CYP3A4 in the metabolic activation of pradefovir. Pradefovir at 0.2, 2, and 20 microM was neither a direct inhibitor nor a mechanism-based inhibitor of CYP3A4, CYP2D6, CYP2C9, CYP2C19, CYP2E1, and CYP1A2 in human liver microsomes. In rats, the liver was the site of metabolic activation of pradefovir, whereas the small intestine did not play a significant role in the metabolic conversion of pradefovir to PMEA. Daily oral dosing (300 mg/kg of body weight) to rats for 8 days showed that pradefovir was not an inducer of P450 enzymes in rats. Furthermore, pradefovir at 10 microg/ml was not an inducer of either CYP1A2 or CYP3A4/5 in primary cultures of human hepatocytes.
authors.affiliation: Drug Development, Valeant Research & Development, Costa Mesa, CA 92626, USA. cclin@valeant.com
authors.forename: Chin-chung
authors.initials: CC
authors.lastname: Lin
authors.forename: Che
authors.initials: C
authors.lastname: Fang
authors.forename: Salete
authors.initials: S
authors.lastname: Benetton
authors.forename: Gui-fen
authors.initials: GF
authors.lastname: Xu
authors.forename: Li-Tain
authors.initials: LT
authors.lastname: Yeh
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors / Q000493:pharmacokinetics / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001711:Biotransformation; D051544:Cytochrome P-450 CYP3A; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics; D004347:Drug Interactions; D004790:Enzyme Induction / Q000187:drug effects; D022781:Hepatocytes / Q000187:drug effects / Q000201:enzymology; D006801:Humans; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000378:metabolism; D007654:Ketoconazole / Q000494:pharmacology; D008862:Microsomes, Liver / Q000378:metabolism; D063065:Organophosphonates / Q000037:antagonists & inhibitors / Q000493:pharmacokinetics* / Q000494:pharmacology; D009943:Organophosphorus Compounds / Q000037:antagonists & inhibitors / Q000493:pharmacokinetics* / Q000494:pharmacology; D011355:Prodrugs; D051381:Rats
chemical_list: D000911:Antibodies, Monoclonal; D065607:Cytochrome P-450 Enzyme Inhibitors; D018076:DNA, Complementary; D007527:Isoenzymes; D063065:Organophosphonates; D009943:Organophosphorus Compounds; D011355:Prodrugs; C053001:adefovir; D003577:Cytochrome P-450 Enzyme System; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; C497721:pradefovir; D000225:Adenine; D007654:Ketoconazole