Abstract
Methodology is described to evaluate the reactivity of acyl glucuronides of carboxylic acid-containing compounds. In this chapter, zomepirac is presented as an example where the reactivity is determined in two complimentary ways: (1) Acyl glucuronides are prepared with UDPGA-fortified microsomes and 1-O-β acyl glucuronide is established based on its sensitivity to hydrolysis by β-glucuronidase. The chemical degradation of 1-O-β isomer in 0.1 M phosphate buffer at pH 7.4 is monitored over time for acyl migration by LC-MS/MS analyses. Half-life of disappearance of 1-O-β acyl glucuronide is used as an index of reactivity. (2) Another index of reactivity of acyl glucuronide is from covalent binding to a protein. Acyl glucuronides are incubated with human serum albumin for 2 h. After pelleting and extensively washing the protein pellets, covalent binding is determined by quantitation of the released aglycone from alkaline hydrolysis.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Fung F, Thornton A, Mybeck M, Hornbuckle K, Muniz E (2001) Evaluation of the characteristics of safety withdrawal of prescription drugs from worldwide pharmaceutical markets—1960–1999. Drug Inf J 35:293–317
Regan SL, Maggs JL, Hammond TG, Lambert C, Williams DP, Park BK (2010) Acyl glucuronides: the good, the bad and the ugly. Biopharm Drug Dispos 31:367–395
FDA (2008) Guidance for industry: safety testing of drug metabolites. http://www.fda.gov/cder/guidance/index.htm
ICH (2010) Guidance for Industry: M3(R2) Nonclinical Safety Studies for the Conduct of Human Clinical Trials and Marketing Authorization for Pharmaceuticals, http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/ucm073246.pdf; Guidance for Industry: M3(R2) Nonclinical Safety Studies for the Conduct of Human Clinical Trials and Marketing Authorization for Pharmaceuticals Questions and Answers(R2), http://www.fda.gov/downloads/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/UCM292340.pdf
Bolze S, Bromet N, Gay-Feutry C, Massiere F, Boulieu R, Hulot T (2002) Development of an in vitro screening model for the biosynthesis of acyl glucuronide metabolites and the assessment of their reactivity toward human serum albumin. Drug Metab Dispos 30(4):404–413
Comble J, Blake JW, Nugent TE, Tobin T (1982) Morphine glucuronide hydrolysis: superiority of β-glucuronidase from patella vulgate. Clin Chem 28(1):83–86
Ebner T, Heinzel G, Prox A, Beschke K, Wachsmuth H (1999) Disposition and chemical stability of telmisartan 1-O-acyl glucuronide. Drug Metab Dispos 27(10):1143–1149
Sawamura R, Okudaira N, Watanabe K, Murai T, Kobayashi Y, Tachibana M, Ohnuki T, Masuda K, Honma H, Kurihara A, Okazaki O (2010) Predictability of idiosyncratic drug toxicity risk for carboxylic acid-containing drugs based on the chemical stability of acyl glucuronide. Drug Metab Dispos 38(10):1857–1864
Acknowledgements
The views expressed here are solely those of the author and do not reflect the opinions of Janssen Research & Development, LLC.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Xu, R.F., Lam, W.W., Chen, J., McMillian, M., Silva, J., Lim, HK. (2014). In Vitro Assessment of the Reactivity of Acyl Glucuronides. In: Caldwell, G., Yan, Z. (eds) Optimization in Drug Discovery. Methods in Pharmacology and Toxicology. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-742-6_30
Download citation
DOI: https://doi.org/10.1007/978-1-62703-742-6_30
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-741-9
Online ISBN: 978-1-62703-742-6
eBook Packages: Springer Protocols