QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: dmd.104.000026v1 32/11/1247    most recent Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by McGinnity, D. F. Articles by Riley, R. J. Search for Related Content PubMed PubMed Citation Articles by McGinnity, D. F. Articles by Riley, R. J.

EVALUATION OF FRESH AND CRYOPRESERVED HEPATOCYTES AS IN VITRO DRUG METABOLISM TOOLS FOR THE PREDICTION OF METABOLIC CLEARANCE

Department of Physical & Metabolic Science, AstraZeneca R&D Charnwood, Leicestershire, United Kingdom

The intrinsic clearances (CL_int) of 50 neutral and basic marketed drugs were determined in fresh human hepatocytes and the data used to predict human in vivo hepatic metabolic clearance (CL_met). A statistically significant correlation between scaled CL_met and actual CL_met was observed (r² = 0.48, p < 0.05), and for 73% of the drugs studied, scaled clearances were within 2-fold of the actual clearance. These data have shown that CL_int data generated in human hepatocytes can be used to provide estimates of human hepatic CL_met for both phase I and phase II processes. In addition, the utility of commercial and in-house cryopreserved hepatocytes was assessed by comparing with data derived from fresh cells. A set of 14 drugs metabolized by the major human cytochromes P450 (P450s) (CYP1A2, 2C9, 2C19, 2D6, and 3A4) and uridine diphosphate glucuronosyltransferases (UGT1A1, 1A4, 1A9, and 2B7) have been used to characterize the activity of freshly isolated and cryopreserved human and dog hepatocytes. The cryopreserved human and dog cells retained on average 94% and 81%, respectively, of the CL_int determined in fresh cells. Cryopreserved hepatocytes retain their full activity for more than 1 year in liquid N₂ and are thus a flexible resource of hepatocytes for in vitro assays. In summary, this laboratory has successfully cryopreserved human and dog hepatocytes as assessed by the turnover of prototypic P450 and UGT substrates, and both fresh and cryopreserved human hepatocytes may be used for the prediction of human hepatic CL_met.

This article has been cited by other articles:

				S. W. Paine, A. J. Parker, P. Gardiner, P. J. H. Webborn, and R. J. Riley Prediction of the Pharmacokinetics of Atorvastatin, Cerivastatin, and Indomethacin Using Kinetic Models Applied to Isolated Rat Hepatocytes Drug Metab. Dispos., July 1, 2008; 36(7): 1365 - 1374. [Abstract] [Full Text] [PDF]

				A. J. Parker and J. B. Houston Rate-Limiting Steps in Hepatic Drug Clearance: Comparison of Hepatocellular Uptake and Metabolism with Microsomal Metabolism of Saquinavir, Nelfinavir, and Ritonavir Drug Metab. Dispos., July 1, 2008; 36(7): 1375 - 1384. [Abstract] [Full Text] [PDF]

				D. F. McGinnity, N. J. Waters, J. Tucker, and R. J. Riley Integrated in Vitro Analysis for the in Vivo Prediction of Cytochrome P450-Mediated Drug-Drug Interactions Drug Metab. Dispos., June 1, 2008; 36(6): 1126 - 1134. [Abstract] [Full Text] [PDF]

				A. Rowland, D. J. Elliot, K. M. Knights, P. I. Mackenzie, and J. O. Miners The "Albumin Effect" and in Vitro-in Vivo Extrapolation: Sequestration of Long-Chain Unsaturated Fatty Acids Enhances Phenytoin Hydroxylation by Human Liver Microsomal and Recombinant Cytochrome P450 2C9 Drug Metab. Dispos., May 1, 2008; 36(5): 870 - 877. [Abstract] [Full Text] [PDF]

				R. Fujiwara, M. Nakajima, H. Yamanaka, M. Katoh, and T. Yokoi Product Inhibition of UDP-Glucuronosyltransferase (UGT) Enzymes by UDP Obfuscates the Inhibitory Effects of UGT Substrates Drug Metab. Dispos., February 1, 2008; 36(2): 361 - 367. [Abstract] [Full Text] [PDF]

				D. Hallifax and J. B. Houston Saturable Uptake of Lipophilic Amine Drugs into Isolated Hepatocytes: Mechanisms and Consequences for Quantitative Clearance Prediction Drug Metab. Dispos., August 1, 2007; 35(8): 1325 - 1332. [Abstract] [Full Text] [PDF]

				M. G. Soars, K. Grime, J. L. Sproston, P. J. H. Webborn, and R. J. Riley Use of Hepatocytes to Assess the Contribution of Hepatic Uptake to Clearance in Vivo Drug Metab. Dispos., June 1, 2007; 35(6): 859 - 865. [Abstract] [Full Text] [PDF]

				Y. Mano, T. Usui, and H. Kamimura Comparison of Inhibition Potentials of Drugs against Zidovudine Glucuronidation in Rat Hepatocytes and Liver Microsomes Drug Metab. Dispos., April 1, 2007; 35(4): 602 - 606. [Abstract] [Full Text] [PDF]

				H. S. Brown, M. Griffin, and J. B. Houston Evaluation of Cryopreserved Human Hepatocytes as an Alternative in Vitro System to Microsomes for the Prediction of Metabolic Clearance Drug Metab. Dispos., February 1, 2007; 35(2): 293 - 301. [Abstract] [Full Text] [PDF]

				D. Hallifax and J. B. Houston Uptake and Intracellular Binding of Lipophilic Amine Drugs by Isolated Rat Hepatocytes and Implications for Prediction of in Vivo Metabolic Clearance Drug Metab. Dispos., November 1, 2006; 34(11): 1829 - 1836. [Abstract] [Full Text] [PDF]

				I. J. Martin, R. J. Lewis, M. A. Bernstein, I. G. Beattie, C. A. Martin, R. J. Riley, and B. Springthorpe Which Hydroxy? Evidence for Species Differences in the Regioselectivity of Glucuronidation in Rat, Dog, and Human in Vitro Systems and Dog in Vivo Drug Metab. Dispos., September 1, 2006; 34(9): 1502 - 1507. [Abstract] [Full Text] [PDF]

				C. Lu, P. Li, R. Gallegos, V. Uttamsingh, C. Q. Xia, G. T. Miwa, S. K. Balani, and L.-S. Gan Comparison of Intrinsic Clearance in Liver Microsomes and Hepatocytes from Rats and Humans: Evaluation of Free Fraction and Uptake in Hepatocytes Drug Metab. Dispos., September 1, 2006; 34(9): 1600 - 1605. [Abstract] [Full Text] [PDF]

				D. F. McGinnity, A. J. Berry, J. R. Kenny, K. Grime, and R. J. Riley EVALUATION OF TIME-DEPENDENT CYTOCHROME P450 INHIBITION USING CULTURED HUMAN HEPATOCYTES Drug Metab. Dispos., August 1, 2006; 34(8): 1291 - 1300. [Abstract] [Full Text] [PDF]

				D. Hallifax, H. C. Rawden, N. Hakooz, and J. B. Houston PREDICTION OF METABOLIC CLEARANCE USING CRYOPRESERVED HUMAN HEPATOCYTES: KINETIC CHARACTERISTICS FOR FIVE BENZODIAZEPINES Drug Metab. Dispos., December 1, 2005; 33(12): 1852 - 1858. [Abstract] [Full Text] [PDF]

				D. F. McGinnity, J. Tucker, S. Trigg, and R. J. Riley PREDICTION OF CYP2C9-MEDIATED DRUG-DRUG INTERACTIONS: A COMPARISON USING DATA FROM RECOMBINANT ENZYMES AND HUMAN HEPATOCYTES Drug Metab. Dispos., November 1, 2005; 33(11): 1700 - 1707. [Abstract] [Full Text] [PDF]

				R. J. Riley, D. F. McGinnity, and R. P. Austin A UNIFIED MODEL FOR PREDICTING HUMAN HEPATIC, METABOLIC CLEARANCE FROM IN VITRO INTRINSIC CLEARANCE DATA IN HEPATOCYTES AND MICROSOMES Drug Metab. Dispos., September 1, 2005; 33(9): 1304 - 1311. [Abstract] [Full Text] [PDF]

				M.-Y. Lee, C. B. Park, J. S. Dordick, and D. S. Clark From the Cover: Metabolizing enzyme toxicology assay chip (MetaChip) for high-throughput microscale toxicity analyses PNAS, January 25, 2005; 102(4): 983 - 987. [Abstract] [Full Text] [PDF]