LC-MS/MS determination of guanabenz E/Z isomers and its application to in vitro and in vivo DMPK profiling studies

Jiashu Xie, Rongrong Jiang, Wei Xie, Bin Cao, Swati S. More

Research output: Contribution to journalArticlepeer-review

Abstract

Endoplasmic reticulum (ER) stress underlies a variety of disorders involving inflammation, such as diabetes, neurodegenerative diseases. Guanabenz acetate (Wytensin®, GA), a clinically approved antihypertensive drug, efficiently counteracts ER stress. The entirety of clinically used GA is the E-isomer, while the Z-isomer is known to lack significant hypotensive properties. We recently discovered that the Z-isomer retains anti-ER stress activity. Coupled with its lack of sedative effects, (Z)-GA is well positioned as a potential therapeutic for a host of ER stress-related disorders. We set forth to characterize the metabolism and pharmacokinetics (DMPK) of (Z)-GA in vitro and in vivo. Toward this end, a reliable and sensitive LC-MS/MS method for simultaneous determination of the (E)- and (Z)-guanabenz was developed. Chromatographic separation of the isomers was achieved on a C18 reverse phase column with a gradient elution. Tandem mass spectrometric detection was conducted using an AB Sciex 5500 QTrap mass spectrometer with positive electrospray ionization. Full validation of the method was performed in mouse plasma with a simple and low plasma volume protein precipitation procedure. The method demonstrated good linearity, reproducibility, and accuracy over a range of 0.5–1000 nM with minimal matrix effect and excellent extraction efficiency. In addition, the developed method was successfully applied to DMPK studies of the GA isomers in vitro and in vivo. Results of these studies revealed for the first time that the DMPK profile of (Z)-guanabenz is distinct from that of (E)-guanabenz, with higher apparent volume of distribution (Vd) and clearance, presumably due to lower plasma protein binding.

Original languageEnglish (US)
Article number114331
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume205
DOIs
StatePublished - Oct 25 2021

Bibliographical note

Funding Information:
We thank the Center for Drug Design at the College of Pharmacy, and a grant from the Winston and Maxine Wallin Neuroscience Discovery fund, University of Minnesota , USA, for supporting this research.

Publisher Copyright:
© 2021 Elsevier B.V.

Keywords

  • (E)-guanabenz
  • (Z)-guanabenz
  • DMPK
  • LC-MS/MS
  • Method validation
  • Protein binding

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