Abstract
Previous studies have shown the importance of the addition of albumin for characterization of hepatic glucuronidation in vitro; however, no reports exist on the effects of albumin on renal or intestinal microsomal glucuronidation assays. This study characterized glucuronidation clearance (CLint, UGT) in human kidney, liver, and intestinal microsomes in the presence and absence of bovine serum albumin (BSA) for seven drugs with differential UDP-glucuronosyltransferase (UGT) 1A9 and UGT2B7 specificity, namely, diclofenac, ezetimibe, gemfibrozil, mycophenolic acid, naloxone, propofol, and telmisartan. The impact of renal CLint, UGT on accuracy of in vitro-in vivo extrapolation (IVIVE) of glucuronidation clearance was investigated. Inclusion of 1% BSA for acidic drugs and 2% for bases/neutral drugs in incubations was found to be suitable for characterization of CLint, UGT in different tissues. Although BSA increased CLint, UGT in all tissues, the extent was tissue- and drug-dependent. Scaled CLint, UGT in the presence of BSA ranged from 2.22 to 207, 0.439 to 24.4, and 0.292 to 23.8 ml · min−1 · g tissue−1 in liver, kidney, and intestinal microsomes. Renal CLint, UGT (per gram of tissue) was up to 2-fold higher in comparison with that for liver for UGT1A9 substrates; in contrast, CLint, UGT for UGT2B7 substrates represented approximately one-third of hepatic estimates. Scaled renal CLint, UGT (in the presence of BSA) was up to 30-fold higher than intestinal glucuronidation for the drugs investigated. Use of in vitro data obtained in the presence of BSA and inclusion of renal clearance improved the IVIVE of glucuronidation clearance, with 50% of drugs predicted within 2-fold of observed values. Characterization and consideration of kidney CLint, UGT is particularly important for UGT1A9 substrates.
Footnotes
The work was funded by a consortium of pharmaceutical companies (GlaxoSmithKline, Lilly, Pfizer, and Servier) within the Centre for Applied Pharmacokinetic Research at the University of Manchester. K.L.G. is a recipient of a Ph.D. studentship from Biotechnology and Biological Sciences Research Council.
Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.
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The online version of this article (available at http://dmd.aspetjournals.org) contains supplemental material.
ABBREVIATIONS:
- UGT
- UDP-glucuronosyltransferase
- FFA
- free fatty acid
- BSA
- bovine serum albumin
- HLM
- human liver microsomes
- IVIVE
- in vitro-in vivo extrapolation
- HKM
- human kidney microsomes
- HIM
- human intestinal microsomes
- MPA
- mycophenolic acid
- LC
- liquid chromatography
- MS/MS
- tandem mass spectrometry.
- Received November 29, 2011.
- Accepted January 23, 2012.
- Copyright © 2012 by The American Society for Pharmacology and Experimental Therapeutics
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