Bennett Ma1, Roy Eisenhandler2, Yuhsin Kuo2, Paul Rearden2, Ying Li3, Peter J Manley4, Sheri Smith2, Karsten Menzel2. 1. Department of Pharmacokinetics, Pharmacodynamics and Drug Metabolism, Merck Research Laboratories, West Point, PA, 19486, USA. bennett_ma@merck.com. 2. Department of Pharmacokinetics, Pharmacodynamics and Drug Metabolism, Merck Research Laboratories, West Point, PA, 19486, USA. 3. Department of Pharmacokinetics, Pharmacodynamics and Drug Metabolism, Merck Research Laboratories, Kenilworth, NJ, USA. 4. Department of Medicinal Chemistry, Merck Research Laboratories, West Point, PA, USA.
Abstract
BACKGROUND AND OBJECTIVES: Prediction of metabolic clearance has been a challenge for compounds exhibiting minimal turnover in typical in vitro stability experiments. The aim of the current study is to evaluate the utilization of plated human hepatocytes to predict intrinsic clearance of low-turnover compounds. METHODS: The disappearance of test compounds was determined for up to 48 h while enzyme activities in plated hepatocytes were monitored concurrently in a complimentary experiment. RESULTS: Consistent with literature reports, marked time-dependent loss of cytochrome P450 (CYP) enzyme activities was observed during the 48-h incubation period. To account for the loss of enzyme activities, a term "fraction of activity remaining" was calculated based on area-under-the-curve derived from the average rate of activity loss (k avg), and then applied as a correction factor for intrinsic clearance determination. Twelve compounds were selected in this study to cover phase I and phase II biotransformation pathways, with in vivo intrinsic clearance values, representing metabolic clearance only, ranging from 0.66 to 47 ml/min/kg. Determination of in vitro intrinsic clearance using three individual preparations of hepatocytes revealed a reasonably good agreement (generally within threefold) between the predicted and the observed metabolic clearance for all 12 compounds tested. CONCLUSIONS: The current results indicated that plated hepatocytes can be utilized to provide clearance predictions for compounds with low-turnover in humans when corrected for the loss in enzyme activities.
BACKGROUND AND OBJECTIVES: Prediction of metabolic clearance has been a challenge for compounds exhibiting minimal turnover in typical in vitro stability experiments. The aim of the current study is to evaluate the utilization of plated human hepatocytes to predict intrinsic clearance of low-turnover compounds. METHODS: The disappearance of test compounds was determined for up to 48 h while enzyme activities in plated hepatocytes were monitored concurrently in a complimentary experiment. RESULTS: Consistent with literature reports, marked time-dependent loss of cytochrome P450 (CYP) enzyme activities was observed during the 48-h incubation period. To account for the loss of enzyme activities, a term "fraction of activity remaining" was calculated based on area-under-the-curve derived from the average rate of activity loss (k avg), and then applied as a correction factor for intrinsic clearance determination. Twelve compounds were selected in this study to cover phase I and phase II biotransformation pathways, with in vivo intrinsic clearance values, representing metabolic clearance only, ranging from 0.66 to 47 ml/min/kg. Determination of in vitro intrinsic clearance using three individual preparations of hepatocytes revealed a reasonably good agreement (generally within threefold) between the predicted and the observed metabolic clearance for all 12 compounds tested. CONCLUSIONS: The current results indicated that plated hepatocytes can be utilized to provide clearance predictions for compounds with low-turnover in humans when corrected for the loss in enzyme activities.
Authors: M J Lamson; J P Sabo; T R MacGregor; J W Pav; L Rowland; A Hawi; M Cappola; P Robinson Journal: Biopharm Drug Dispos Date: 1999-09 Impact factor: 1.627
Authors: Cornelis E C A Hop; Mark J Cole; Ralph E Davidson; David B Duignan; James Federico; John S Janiszewski; Kelly Jenkins; Suzanne Krueger; Rebecca Lebowitz; Theodore E Liston; Walter Mitchell; Mark Snyder; Stefan J Steyn; John R Soglia; Christine Taylor; Matt D Troutman; John Umland; Michael West; Kevin M Whalen; Veronica Zelesky; Sabrina X Zhao Journal: Curr Drug Metab Date: 2008-11 Impact factor: 3.731
Authors: Tom S Chan; Hongbin Yu; Amanda Moore; Salman R Khetani; Salman R Kehtani; Donald Tweedie Journal: Drug Metab Dispos Date: 2013-08-19 Impact factor: 3.922
Authors: Britta Burkhardt; Juan José Martinez-Sanchez; Anastasia Bachmann; Ruth Ladurner; Andreas K Nüssler Journal: Hepatol Int Date: 2013-11-21 Impact factor: 6.047
Authors: Cornelia M Smith; Christina K Nolan; Manda A Edwards; Jean B Hatfield; Todd W Stewart; Stephen S Ferguson; Edward L Lecluyse; Jasminder Sahi Journal: J Pharm Sci Date: 2012-07-17 Impact factor: 3.534
Authors: Antonio F Hernandez-Jerez; Paulien Adriaanse; Annette Aldrich; Philippe Berny; Tamara Coja; Sabine Duquesne; Andreas Focks; Marina Marinovich; Maurice Millet; Olavi Pelkonen; Silvia Pieper; Aaldrik Tiktak; Christopher J Topping; Anneli Widenfalk; Martin Wilks; Gerrit Wolterink; Ursula Gundert-Remy; Jochem Louisse; Serge Rudaz; Emanuela Testai; Alfonso Lostia; Jean-Lou Dorne; Juan Manuel Parra Morte Journal: EFSA J Date: 2021-12-23
Authors: Marjory Moreau; Pankajini Mallick; Marci Smeltz; Saad Haider; Chantel I Nicolas; Salil N Pendse; Jeremy A Leonard; Matthew W Linakis; Patrick D McMullen; Rebecca A Clewell; Harvey J Clewell; Miyoung Yoon Journal: Front Toxicol Date: 2022-04-29