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

Ravinder Earla, 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.
Santosh Kumar, 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.
Lei Wang, 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.).
Steven Bosinger, 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.).
Junhao Li, 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.).
Ankit Shah, 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.).
Mohitkumar Gangwani, 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.).
Anantha Nookala, 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.).
Xun Liu, 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.).
Lu Cao, 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.).
Austin Jackson, 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.).
Peter S. Silverstein, 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.).
Howard S. Fox, 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.).
Weihua Li, 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.).
Anil Kumar, 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.).

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.

Publication Title

Drug metabolism and disposition: the biological fate of chemicals

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