title: The influence of nonspecific microsomal binding on apparent intrinsic clearance, and its prediction from physicochemical properties.
abstract: The apparent intrinsic clearance of 13 drugs has been determined using rat liver microsomes at three different concentrations of microsomal protein. The kinetics was studied using the in vitro half-life method. The nonspecific binding of these drugs to the microsomes was also studied under the same conditions, except for cofactor removal, using equilibrium dialysis. The intrinsic clearances are shown to be dependent on the microsomal concentration, but are approximately constant when corrected for the extent of nonspecific binding to the microsomes. The large difference between observed intrinsic clearance and unbound intrinsic clearance that exists for some compounds, particularly lipophilic bases, is highlighted. A simple model has been developed for understanding the binding of compounds to microsomes and is demonstrated to accurately predict the extent of microsomal binding at one concentration of microsomes from measurement at another. The binding of a further 25 drugs to rat liver microsomes at a microsomal concentration of 1 mg/ml was also studied, along with measurements of lipophilicity using octanol-water partition coefficients. It is shown that the extent of microsomal binding is correlated with lipophilicity, but that basic compounds show a different behavior to acidic and neutral compounds. Microsomal binding is shown to be best predicted using a model where log P is used for basic compounds, and log D(7.4) is used for acidic and neutral compounds. This model has been developed further so that the extent of binding to microsomes of any given concentration can be estimated purely from a knowledge of lipophilicity and ionization.
authors.affiliation: Department of Physical and Metabolic Science, AstraZeneca R&D Charnwood, Loughborough, United Kingdom. rupert.austin@astrazeneca.com
authors.forename: Rupert P
authors.initials: RP
authors.lastname: Austin
authors.forename: Patrick
authors.initials: P
authors.lastname: Barton
authors.forename: Scott L
authors.initials: SL
authors.lastname: Cockroft
authors.forename: Mark C
authors.initials: MC
authors.lastname: Wenlock
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Riley
mesh_terms: D000818:Animals; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D008297:Male; D008657:Metabolic Clearance Rate / Q000502:physiology; D008862:Microsomes, Liver / Q000737:chemistry / Q000378:metabolism*; D008956:Models, Chemical; D004364:Pharmaceutical Preparations / Q000737:chemistry / Q000378:metabolism*; D011237:Predictive Value of Tests; D011485:Protein Binding / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D004364:Pharmaceutical Preparations