title: Enhanced methamphetamine metabolism in rhesus macaque as compared with human: an analysis using a novel method of liquid chromatography with tandem mass spectrometry, kinetic study, and substrate docking.
abstract: Methamphetamine (MA), which remains one of the widely used drugs of abuse, is metabolized by the cytochrome P450 (P450) family of enzymes in humans. However, metabolism of methamphetamine in macaques is poorly understood. Therefore, we first developed and validated a very sensitive liquid chromatography with tandem mass spectrometry (LC-MS/MS) method using solid phase extraction of rhesus plasma with a lower limit of quantitation at 1.09 ng/ml for MA and its metabolites, 4-hydroxy methamphetamine (4-OH MA), amphetamine (AM), 4-OH amphetamine (4-OH AM), and norephedrine. We then analyzed plasma samples of MA-treated rhesus, which showed >10-fold higher concentrations of AM (∼29 ng/ml) and 4-OH AM (∼28 ng/ml) than MA (∼2 ng/ml). Because the plasma levels of MA metabolites in rhesus were much higher than in human samples, we examined MA metabolism in human and rhesus microsomes. Interestingly, the results showed that AM and 4-OH AM were formed more rapidly and that the catalytic efficiency (Vmax/Km) for the formation of AM was ∼8-fold higher in rhesus than in human microsomes. We further examined the differences in these kinetic characteristics using three selective inhibitors of each human CYP2D6 and CYP3A4 enzymes. The results showed that each of these inhibitors inhibited both d- and l-MA metabolism by 20%-60% in human microsomes but not in rhesus microsomes. The differences between human and rhesus CYP2D6 and CYP3A4 enzymes were further assessed by docking studies for both d and l-MA. In conclusion, our results demonstrated an enhanced MA metabolism in rhesus compared with humans, which is likely to be caused by differences in MA-metabolizing P450 enzymes between these species.
authors.affiliation: Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri (R.E., A.S., M.K.G., A.N., X.L., L.C., A.J., P.S.S., A.K.); Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Sciences, Memphis, Tennessee (S.K.); Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, People's Republic of China (L.W., J.L., W.L.); Yerkes National Primate Research Center, Emory University, Atlanta, Georgia (S.B.); University of Nebraska Medical Center, Omaha, Nebraska (H.S.F.) earlar@umkc.edu ksantosh@uthsc.edu.
authors.forename: Ravinder
authors.initials: R
authors.lastname: Earla
authors.affiliation: Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri (R.E., A.S., M.K.G., A.N., X.L., L.C., A.J., P.S.S., A.K.); Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Sciences, Memphis, Tennessee (S.K.); Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, People's Republic of China (L.W., J.L., W.L.); Yerkes National Primate Research Center, Emory University, Atlanta, Georgia (S.B.); University of Nebraska Medical Center, Omaha, Nebraska (H.S.F.) earlar@umkc.edu ksantosh@uthsc.edu.
authors.forename: Santosh
authors.initials: S
authors.lastname: Kumar
authors.affiliation: Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri (R.E., A.S., M.K.G., A.N., X.L., L.C., A.J., P.S.S., A.K.); Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Sciences, Memphis, Tennessee (S.K.); Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, People's Republic of China (L.W., J.L., W.L.); Yerkes National Primate Research Center, Emory University, Atlanta, Georgia (S.B.); University of Nebraska Medical Center, Omaha, Nebraska (H.S.F.).
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.affiliation: Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri (R.E., A.S., M.K.G., A.N., X.L., L.C., A.J., P.S.S., A.K.); Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Sciences, Memphis, Tennessee (S.K.); Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, People's Republic of China (L.W., J.L., W.L.); Yerkes National Primate Research Center, Emory University, Atlanta, Georgia (S.B.); University of Nebraska Medical Center, Omaha, Nebraska (H.S.F.).
authors.forename: Steven
authors.initials: S
authors.lastname: Bosinger
authors.affiliation: Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri (R.E., A.S., M.K.G., A.N., X.L., L.C., A.J., P.S.S., A.K.); Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Sciences, Memphis, Tennessee (S.K.); Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, People's Republic of China (L.W., J.L., W.L.); Yerkes National Primate Research Center, Emory University, Atlanta, Georgia (S.B.); University of Nebraska Medical Center, Omaha, Nebraska (H.S.F.).
authors.forename: Junhao
authors.initials: J
authors.lastname: Li
authors.affiliation: Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri (R.E., A.S., M.K.G., A.N., X.L., L.C., A.J., P.S.S., A.K.); Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Sciences, Memphis, Tennessee (S.K.); Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, People's Republic of China (L.W., J.L., W.L.); Yerkes National Primate Research Center, Emory University, Atlanta, Georgia (S.B.); University of Nebraska Medical Center, Omaha, Nebraska (H.S.F.).
authors.forename: Ankit
authors.initials: A
authors.lastname: Shah
authors.affiliation: Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri (R.E., A.S., M.K.G., A.N., X.L., L.C., A.J., P.S.S., A.K.); Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Sciences, Memphis, Tennessee (S.K.); Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, People's Republic of China (L.W., J.L., W.L.); Yerkes National Primate Research Center, Emory University, Atlanta, Georgia (S.B.); University of Nebraska Medical Center, Omaha, Nebraska (H.S.F.).
authors.forename: Mohitkumar
authors.initials: M
authors.lastname: Gangwani
authors.affiliation: Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri (R.E., A.S., M.K.G., A.N., X.L., L.C., A.J., P.S.S., A.K.); Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Sciences, Memphis, Tennessee (S.K.); Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, People's Republic of China (L.W., J.L., W.L.); Yerkes National Primate Research Center, Emory University, Atlanta, Georgia (S.B.); University of Nebraska Medical Center, Omaha, Nebraska (H.S.F.).
authors.forename: Anantha
authors.initials: A
authors.lastname: Nookala
authors.affiliation: Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri (R.E., A.S., M.K.G., A.N., X.L., L.C., A.J., P.S.S., A.K.); Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Sciences, Memphis, Tennessee (S.K.); Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, People's Republic of China (L.W., J.L., W.L.); Yerkes National Primate Research Center, Emory University, Atlanta, Georgia (S.B.); University of Nebraska Medical Center, Omaha, Nebraska (H.S.F.).
authors.forename: Xun
authors.initials: X
authors.lastname: Liu
authors.affiliation: Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri (R.E., A.S., M.K.G., A.N., X.L., L.C., A.J., P.S.S., A.K.); Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Sciences, Memphis, Tennessee (S.K.); Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, People's Republic of China (L.W., J.L., W.L.); Yerkes National Primate Research Center, Emory University, Atlanta, Georgia (S.B.); University of Nebraska Medical Center, Omaha, Nebraska (H.S.F.).
authors.forename: Lu
authors.initials: L
authors.lastname: Cao
authors.affiliation: Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri (R.E., A.S., M.K.G., A.N., X.L., L.C., A.J., P.S.S., A.K.); Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Sciences, Memphis, Tennessee (S.K.); Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, People's Republic of China (L.W., J.L., W.L.); Yerkes National Primate Research Center, Emory University, Atlanta, Georgia (S.B.); University of Nebraska Medical Center, Omaha, Nebraska (H.S.F.).
authors.forename: Austin
authors.initials: A
authors.lastname: Jackson
authors.affiliation: Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri (R.E., A.S., M.K.G., A.N., X.L., L.C., A.J., P.S.S., A.K.); Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Sciences, Memphis, Tennessee (S.K.); Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, People's Republic of China (L.W., J.L., W.L.); Yerkes National Primate Research Center, Emory University, Atlanta, Georgia (S.B.); University of Nebraska Medical Center, Omaha, Nebraska (H.S.F.).
authors.forename: Peter S
authors.initials: PS
authors.lastname: Silverstein
authors.affiliation: Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri (R.E., A.S., M.K.G., A.N., X.L., L.C., A.J., P.S.S., A.K.); Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Sciences, Memphis, Tennessee (S.K.); Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, People's Republic of China (L.W., J.L., W.L.); Yerkes National Primate Research Center, Emory University, Atlanta, Georgia (S.B.); University of Nebraska Medical Center, Omaha, Nebraska (H.S.F.).
authors.forename: Howard S
authors.initials: HS
authors.lastname: Fox
authors.affiliation: Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri (R.E., A.S., M.K.G., A.N., X.L., L.C., A.J., P.S.S., A.K.); Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Sciences, Memphis, Tennessee (S.K.); Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, People's Republic of China (L.W., J.L., W.L.); Yerkes National Primate Research Center, Emory University, Atlanta, Georgia (S.B.); University of Nebraska Medical Center, Omaha, Nebraska (H.S.F.).
authors.forename: Weihua
authors.initials: W
authors.lastname: Li
authors.affiliation: Division of Pharmacology and Toxicology, School of Pharmacy, University of Missouri-Kansas City, Kansas City, Missouri (R.E., A.S., M.K.G., A.N., X.L., L.C., A.J., P.S.S., A.K.); Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Sciences, Memphis, Tennessee (S.K.); Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, People's Republic of China (L.W., J.L., W.L.); Yerkes National Primate Research Center, Emory University, Atlanta, Georgia (S.B.); University of Nebraska Medical Center, Omaha, Nebraska (H.S.F.).
authors.forename: Anil
authors.initials: A
authors.lastname: Kumar
mesh_terms: D000661:Amphetamine / Q000097:blood* / Q000378:metabolism*; D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D019389:Cytochrome P-450 CYP2D6 / Q000378:metabolism; D051544:Cytochrome P-450 CYP3A / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008253:Macaca mulatta / Q000378:metabolism; D008694:Methamphetamine / Q000097:blood* / Q000378:metabolism*; D008862:Microsomes, Liver / Q000378:metabolism; D010665:Phenylpropanolamine / Q000097:blood* / Q000378:metabolism*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D010665:Phenylpropanolamine; D008694:Methamphetamine; D000661:Amphetamine; D019389:Cytochrome P-450 CYP2D6; D051544:Cytochrome P-450 CYP3A