title: On the maintenance of hepatocyte intracellular pH 7.0 in the in-vitro metabolic stability assay.
abstract: The account of pH difference between hepatocytes (intracellular pH 7.0) and extracellular water (pH 7.4) leads to the novel equation for hepatic clearance (Berezhkovskiy, J Pharma Sci 100:1167-1683, 2011). The metabolic stability assay using hepatocytes is commonly performed in the incubation buffer of pH 7.4. If hepatocytes retain their physiological pH 7.0 in these conditions, then the assay would mimic the in vivo condition, that is pH 7.4 for plasma and extracellular water, and pH 7.0 in hepatocytes. In this case the rate of drug elimination, taken as proportional to unbound drug concentration in buffer, would correspond to the in vivo rate of drug elimination as proportional to the unbound drug concentration in the extracellular water. Consequently the commonly used PBPK equation for the rate of hepatic elimination, and the equation for hepatic clearance would be valid. However, the experiment designed to determine hepatocyte internal pH indicated that it was not maintained in the in vitro stability assay, so that hepatocytes acquire the same pH as the incubation buffer. Thus, the novel equations for hepatic clearance (that include an ionization factor) should be applied regardless if the intrinsic clearance was obtained either from microsomal or hepatocyte stability assay.
authors.affiliation: Genentech Inc., 1 DNA Way, South San Francisco, CA, 94080, USA, leo.berezh@yahoo.com.
authors.forename: Leonid M
authors.initials: LM
authors.lastname: Berezhkovskiy
authors.forename: Susan
authors.initials: S
authors.lastname: Wong
authors.forename: Jason S
authors.initials: JS
authors.lastname: Halladay
mesh_terms: D000818:Animals; D004285:Dogs; D045604:Extracellular Fluid / Q000378:metabolism*; D006207:Half-Life; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008657:Metabolic Clearance Rate; D051379:Mice; D008954:Models, Biological; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D010599:Pharmacokinetics; D051381:Rats; D014018:Tissue Distribution
chemical_list: D004364:Pharmaceutical Preparations