RT Journal Article SR Electronic T1 Influence of Microsomal Concentration on Apparent Intrinsic Clearance: Implications for Scaling in Vitro Data JF Drug Metabolism and Disposition JO Drug Metab Dispos FD American Society for Pharmacology and Experimental Therapeutics SP 1332 OP 1336 VO 29 IS 10 A1 J. Cory Kalvass A1 David A. Tess A1 Craig Giragossian A1 Michael C. Linhares A1 Tristan S. Maurer YR 2001 UL http://dmd.aspetjournals.org/content/29/10/1332.abstract AB The influence of microsomal concentration on unbound fraction (fumic), half-life (t1/2), apparent intrinsic clearance (CLint,app) and apparent Michaelis-Menten constant (Km,app) was examined for two compounds, one representative of high nonspecific binding to microsomes (compound A) and one representative of low (compound B). Kinetic parameters were estimated for the two probe compounds at two human microsomal protein concentrations (0.46 and 2.3 mg/ml) and cytochrome P450 concentrations (0.20 and 1.0 μM), representing a 5-fold difference in microsomal concentration. For compound A, fumic and CLint,app were inversely proportional to microsomal concentration. Conversely, theKm,app of compound A was proportional to microsomal concentration and the half-life was unchanged. For compound B, half-life was inversely proportional to microsomal concentration. In this case, fumic, CLint,app, andKm,app were not proportionally influenced. The experimental observations were entirely consistent with that predicted by a mathematical relationship between microsomal concentration, fumic, t1/2, CLint,app, and Km,app. These results demonstrate that when nonspecific binding is extensive, CLint,app is dependent on the arbitrary choice of microsomal concentration included in the incubation. The American Society for Pharmacology and Experimental Therapeutics