Activation and deactivation of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine by cytochrome P450 enzymes and flavin-containing monooxygenases in common marmosets (Callithrix jacchus).
The potential proneurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induces Parkinson-like syndromes in common marmosets, other primates, and humans. MPTP is metabolically activated to 1-methyl-4-phenyl-2,3-dihydropyridinium and 1-methyl-4-phenylpyridinium ions (MPDP(+) and MPP(+), respectively) by desaturation reactions. MPTP is deactivated to 4-phenyl-1,2,3,6-tetrahydropyridine (PTP) by N-demethylation and is also deactivated to MPTP N-oxide. The roles of cytochrome P450 (P450) enzymes and flavin-containing monooxygenases (FMOs) in the oxidative metabolism of MPTP-treated marmosets are not yet fully clarified. This study aimed to elucidate P450- and FMO-dependent MPTP metabolism in marmoset liver and brain. Rates of MPTP N-oxygenation in liver microsomes were similar to those in brain microsomes from 11 individual marmosets (substrate concentration, 50 μM) and were correlated with rates of benzydamine N-oxygenation (r = 0.75, P < 0.05); the reactions were inhibited by methimazole (10 μM). MPTP N-oxygenation was efficiently mediated by recombinantly expressed marmoset FMO3. Rates of PTP formation by MPTP N-demethylation in marmoset liver microsomes were correlated with bufuralol 1'-hydroxylation rates (r = 0.77, P < 0.01) and were suppressed by quinidine (1 μM), thereby indicating the importance of marmoset CYP2D6 in PTP formation. MPTP transformations to MPDP(+) and MPP(+) were efficiently catalyzed by recombinant marmoset CYP2D6 and human CYP1A2. These results indicated the contributions of multiple drug-metabolizing enzymes to MPTP oxidation, especially marmoset FMO3 in deactivation (N-oxygenation) and marmoset CYP2D6 for both MPTP deactivation and MPTP activation to MPDP(+) and MPP(+). These findings provide a foundation for understanding MPTP metabolism and for the successful production of preclinical marmoset models.
Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida, Tokyo, Japan (S.U., N.M., M.S., H.Y.); Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Wakayama, Japan (Y.U.); Department of Applied Developmental Biology, Central Institute for Experimental Animals, Kawasaki, Kanagawa, Japan (T.I., E.S.); and Keio Advanced Research Center, Keio University, Minato-ku, Tokyo, Japan (E.S.).
Shotaro
S
Uehara
Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida, Tokyo, Japan (S.U., N.M., M.S., H.Y.); Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Wakayama, Japan (Y.U.); Department of Applied Developmental Biology, Central Institute for Experimental Animals, Kawasaki, Kanagawa, Japan (T.I., E.S.); and Keio Advanced Research Center, Keio University, Minato-ku, Tokyo, Japan (E.S.).
Yasuhiro
Y
Uno
Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida, Tokyo, Japan (S.U., N.M., M.S., H.Y.); Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Wakayama, Japan (Y.U.); Department of Applied Developmental Biology, Central Institute for Experimental Animals, Kawasaki, Kanagawa, Japan (T.I., E.S.); and Keio Advanced Research Center, Keio University, Minato-ku, Tokyo, Japan (E.S.).
Takashi
T
Inoue
Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida, Tokyo, Japan (S.U., N.M., M.S., H.Y.); Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Wakayama, Japan (Y.U.); Department of Applied Developmental Biology, Central Institute for Experimental Animals, Kawasaki, Kanagawa, Japan (T.I., E.S.); and Keio Advanced Research Center, Keio University, Minato-ku, Tokyo, Japan (E.S.).
Norie
N
Murayama
Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida, Tokyo, Japan (S.U., N.M., M.S., H.Y.); Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Wakayama, Japan (Y.U.); Department of Applied Developmental Biology, Central Institute for Experimental Animals, Kawasaki, Kanagawa, Japan (T.I., E.S.); and Keio Advanced Research Center, Keio University, Minato-ku, Tokyo, Japan (E.S.).
Makiko
M
Shimizu
Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida, Tokyo, Japan (S.U., N.M., M.S., H.Y.); Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Wakayama, Japan (Y.U.); Department of Applied Developmental Biology, Central Institute for Experimental Animals, Kawasaki, Kanagawa, Japan (T.I., E.S.); and Keio Advanced Research Center, Keio University, Minato-ku, Tokyo, Japan (E.S.).
Erika
E
Sasaki
Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida, Tokyo, Japan (S.U., N.M., M.S., H.Y.); Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Wakayama, Japan (Y.U.); Department of Applied Developmental Biology, Central Institute for Experimental Animals, Kawasaki, Kanagawa, Japan (T.I., E.S.); and Keio Advanced Research Center, Keio University, Minato-ku, Tokyo, Japan (E.S.) hyamazak@ac.shoyaku.ac.jp.
Hiroshi
H
Yamazaki
D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine / Q000031:analogs & derivatives / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000378:metabolism; D002144:Callithrix / Q000378:metabolism*; D019388:Cytochrome P-450 CYP1A2 / Q000378:metabolism; D019389:Cytochrome P-450 CYP2D6 / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004138:Dinitrocresols / Q000378:metabolism*; D005260:Female; D006900:Hydroxylation; D008658:Inactivation, Metabolic / Q000502:physiology; D008099:Liver / Q000378:metabolism; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D010084:Oxidation-Reduction; D010105:Oxygenases / Q000378:metabolism
D004138:Dinitrocresols; C040846:4-phenyl-1,2,3,6-tetrahydropyridine; C024132:4,6-dinitro-o-cresol; D003577:Cytochrome P-450 Enzyme System; D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; D006899:Mixed Function Oxygenases; D010105:Oxygenases; C020066:dimethylaniline monooxygenase (N-oxide forming); D019388:Cytochrome P-450 CYP1A2; D019389:Cytochrome P-450 CYP2D6