Metabolic Profiling of a CB2 Agonist, AM9338, Using LC-MS and Microcoil-NMR: Identification of a Novel Dihydroxy Adamantyl Metabolite
Adamantyl groups are key structural subunit commonly used in many marketed drugs targeting diseases ranging from viral infections to neurological disorders. The metabolic disposition of adamantyl compounds has been mostly studied using LC-MS based approaches. However, metabolite quantities isolated...
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doaj-2cab6af2fb124e13b6733cb3eb4432f82020-11-25T02:11:20ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122020-09-011110.3389/fphar.2020.575691575691Metabolic Profiling of a CB2 Agonist, AM9338, Using LC-MS and Microcoil-NMR: Identification of a Novel Dihydroxy Adamantyl MetaboliteChandrashekhar Honrao0Xiaoyu Ma1Shashank Kulkarni2Vinit Joshi3Michael Malamas4Alexander Zvonok5JodiAnne Wood6David Strand7Jason J. Guo8Jason J. Guo9Jason J. Guo10Alexandros Makriyannis11Alexandros Makriyannis12Alexandros Makriyannis13Center for Drug Discovery and Department of Pharmaceutical Sciences, Northeastern University, Boston, MA, United StatesCenter for Drug Discovery and Department of Pharmaceutical Sciences, Northeastern University, Boston, MA, United StatesCenter for Drug Discovery and Department of Pharmaceutical Sciences, Northeastern University, Boston, MA, United StatesCenter for Drug Discovery and Department of Pharmaceutical Sciences, Northeastern University, Boston, MA, United StatesCenter for Drug Discovery and Department of Pharmaceutical Sciences, Northeastern University, Boston, MA, United StatesMAK Scientific LLC, Burlington, MA, United StatesCenter for Drug Discovery and Department of Pharmaceutical Sciences, Northeastern University, Boston, MA, United StatesProtasis Corporation, Seabrook, NH, United StatesCenter for Drug Discovery and Department of Pharmaceutical Sciences, Northeastern University, Boston, MA, United StatesBarnett Institute for Chemical and Biological Analysis, Northeastern University, Boston, MA, United StatesDepartment of Chemistry and Chemical Biology, Northeastern University, Boston, MA, United StatesCenter for Drug Discovery and Department of Pharmaceutical Sciences, Northeastern University, Boston, MA, United StatesMAK Scientific LLC, Burlington, MA, United StatesDepartment of Chemistry and Chemical Biology, Northeastern University, Boston, MA, United StatesAdamantyl groups are key structural subunit commonly used in many marketed drugs targeting diseases ranging from viral infections to neurological disorders. The metabolic disposition of adamantyl compounds has been mostly studied using LC-MS based approaches. However, metabolite quantities isolated from biological preparations are often insufficient for unambiguous structural characterization by NMR. In this work, we utilized microcoil NMR in conjunction with LC-MS to characterize liver microsomal metabolites of an adamantyl based CB2 agonist AM9338, 1-(3-(1H-1,2,3-triazol-1-yl) propyl)-N-(adamantan-1-yl)-1H-indazole-3-carboxamide, a candidate compound for potential multiple sclerosis treatment. We have identified a total of 9 oxidative metabolites of AM9338 whereas mono- or di-hydroxylation of the adamantyl moiety is the primary metabolic pathway. While it is generally believed that the tertiary adamantyl carbons are the preferred sites of CYP450 oxidation, both the mono- and di-hydroxyl metabolites of AM9338 show that the primary oxidative sites are located on the secondary adamantyl carbons. To our knowledge this di-hydroxylated metabolite is a novel adamantyl metabolite that has not been reported before. Further, the stereochemistry of both mono- and di-hydroxyl adamantyl metabolites has been determined using NOE correlations. Furthermore, docking of AM9338 into the CYP3A4 metabolic enzyme corroborates with our experimental findings, and the modelling results also provide a possible mechanism for the unusual susceptibility of adamantyl secondary carbons to metabolic oxidations. The novel dihydroxylated AM9338 metabolite identified in this study, along with the previously known adamantyl metabolites, gives a more complete picture of the metabolic disposition for adamantyl compounds.https://www.frontiersin.org/article/10.3389/fphar.2020.575691/fullmetabolite identificationLC-MSmicro-coil NMRadamantyl metabolismdi-hydroxyl adamantyl metaboliteCYP3A4 metabolism |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Chandrashekhar Honrao Xiaoyu Ma Shashank Kulkarni Vinit Joshi Michael Malamas Alexander Zvonok JodiAnne Wood David Strand Jason J. Guo Jason J. Guo Jason J. Guo Alexandros Makriyannis Alexandros Makriyannis Alexandros Makriyannis |
spellingShingle |
Chandrashekhar Honrao Xiaoyu Ma Shashank Kulkarni Vinit Joshi Michael Malamas Alexander Zvonok JodiAnne Wood David Strand Jason J. Guo Jason J. Guo Jason J. Guo Alexandros Makriyannis Alexandros Makriyannis Alexandros Makriyannis Metabolic Profiling of a CB2 Agonist, AM9338, Using LC-MS and Microcoil-NMR: Identification of a Novel Dihydroxy Adamantyl Metabolite Frontiers in Pharmacology metabolite identification LC-MS micro-coil NMR adamantyl metabolism di-hydroxyl adamantyl metabolite CYP3A4 metabolism |
author_facet |
Chandrashekhar Honrao Xiaoyu Ma Shashank Kulkarni Vinit Joshi Michael Malamas Alexander Zvonok JodiAnne Wood David Strand Jason J. Guo Jason J. Guo Jason J. Guo Alexandros Makriyannis Alexandros Makriyannis Alexandros Makriyannis |
author_sort |
Chandrashekhar Honrao |
title |
Metabolic Profiling of a CB2 Agonist, AM9338, Using LC-MS and Microcoil-NMR: Identification of a Novel Dihydroxy Adamantyl Metabolite |
title_short |
Metabolic Profiling of a CB2 Agonist, AM9338, Using LC-MS and Microcoil-NMR: Identification of a Novel Dihydroxy Adamantyl Metabolite |
title_full |
Metabolic Profiling of a CB2 Agonist, AM9338, Using LC-MS and Microcoil-NMR: Identification of a Novel Dihydroxy Adamantyl Metabolite |
title_fullStr |
Metabolic Profiling of a CB2 Agonist, AM9338, Using LC-MS and Microcoil-NMR: Identification of a Novel Dihydroxy Adamantyl Metabolite |
title_full_unstemmed |
Metabolic Profiling of a CB2 Agonist, AM9338, Using LC-MS and Microcoil-NMR: Identification of a Novel Dihydroxy Adamantyl Metabolite |
title_sort |
metabolic profiling of a cb2 agonist, am9338, using lc-ms and microcoil-nmr: identification of a novel dihydroxy adamantyl metabolite |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Pharmacology |
issn |
1663-9812 |
publishDate |
2020-09-01 |
description |
Adamantyl groups are key structural subunit commonly used in many marketed drugs targeting diseases ranging from viral infections to neurological disorders. The metabolic disposition of adamantyl compounds has been mostly studied using LC-MS based approaches. However, metabolite quantities isolated from biological preparations are often insufficient for unambiguous structural characterization by NMR. In this work, we utilized microcoil NMR in conjunction with LC-MS to characterize liver microsomal metabolites of an adamantyl based CB2 agonist AM9338, 1-(3-(1H-1,2,3-triazol-1-yl) propyl)-N-(adamantan-1-yl)-1H-indazole-3-carboxamide, a candidate compound for potential multiple sclerosis treatment. We have identified a total of 9 oxidative metabolites of AM9338 whereas mono- or di-hydroxylation of the adamantyl moiety is the primary metabolic pathway. While it is generally believed that the tertiary adamantyl carbons are the preferred sites of CYP450 oxidation, both the mono- and di-hydroxyl metabolites of AM9338 show that the primary oxidative sites are located on the secondary adamantyl carbons. To our knowledge this di-hydroxylated metabolite is a novel adamantyl metabolite that has not been reported before. Further, the stereochemistry of both mono- and di-hydroxyl adamantyl metabolites has been determined using NOE correlations. Furthermore, docking of AM9338 into the CYP3A4 metabolic enzyme corroborates with our experimental findings, and the modelling results also provide a possible mechanism for the unusual susceptibility of adamantyl secondary carbons to metabolic oxidations. The novel dihydroxylated AM9338 metabolite identified in this study, along with the previously known adamantyl metabolites, gives a more complete picture of the metabolic disposition for adamantyl compounds. |
topic |
metabolite identification LC-MS micro-coil NMR adamantyl metabolism di-hydroxyl adamantyl metabolite CYP3A4 metabolism |
url |
https://www.frontiersin.org/article/10.3389/fphar.2020.575691/full |
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