Synthesis of 13C215N2‐labeled anti‐inflammatory and cytoprotective tricyclic bis(cyanoenone) ([13C215N2]‐TBE‐31) as an internal standard for quantification by stable isotope dilution LC‐MS method |
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| Authors: | Suqing Zheng Jeffrey T.‐J. Huang Elena V. Knatko Sheila Sharp Maureen Higgins Iwao Ojima Albena T. Dinkova‐Kostova Tadashi Honda |
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| Affiliation: | 1. Institute of Chemical Biology and Drug Discovery, Stony Brook University, Stony Brook, NY, USA;2. Jacqui Wood Cancer Centre, Division of Cancer Research, Medical Research Institute, University of Dundee, Dundee, Scotland, UK;3. Department of Chemistry, Stony Brook University, Stony Brook, NY, USA;4. Lewis B. and Dorothy Cullman Cancer Chemoprotection Center, Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD, USA |
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| Abstract: | Tricyclic bis(cyanoenone), TBE‐31, one of the most potent activators of the Keap1/Nrf2/antioxidant response element pathway, has been developed as a new anti‐inflammatory and cytoprotective agent. 13C215N2‐labeled TBE‐31 ([13C215N2]‐TBE‐31), which has two 13C and two 15N atoms in two cyano groups, was designed to develop a method for quantification of cell, tissue, and plasma levels of TBE‐31 that involves chromatography/mass spectrometry coupled with the use of a stable isotope‐labeled internal standard. [13C215N2]‐TBE‐31 was successfully synthesized in four steps from a previously reported intermediate, which is prepared in 11 steps from cyclohexanone, by introduction of two 13C atoms with ethyl [13C]formate and two 15N atoms with hydroxyl[15N]amine. The stable isotope dilution liquid chromatography–mass spectrometry method for quantification of TBE‐31 was successfully developed using [13C215N2]‐TBE‐31 to compensate for any variables encountered during sample processing and analysis. |
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| Keywords: | isotopically labeled synthesis carbon‐13 nitrogen‐15 stable isotope dilution LC‐MS method Keap1/Nrf2/ARE activator anti‐inflammatory cytoprotective |
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