Renal transport kinetics of chlorothiazide in the isolated perfused rat kidney

The effect of protein binding on the renal excretion of chlorothiazide was examined in the isolated perfused rat kidney. Drug studies were performed in three rats at 6.00% bovine serum albumin + 0% dextran and in three rats at 0.25% bovine serum albumin + 3.83% dextran. Chlorothiazide was introduced into the recirculating perfusate at an initial concentration of 100 micrograms/ml, and was assayed using high-performance liquid chromatography. Functional viability of the kidney was assessed by measuring the fractional excretion of sodium and glucose, and the glomerular filtration rate. The protein binding of drug in perfusate was determined by equilibrium dialysis. These experimental conditions resulted in an approximate 14-fold increase of percent free (from 5.3-72.0%), and a 3-fold increase of renal (from 1.03-3.30 ml/min) and secretion (from 1.01-2.83 ml/min) clearances. The data were best fitted by a model in which one Michaelis-Menten term was used to describe active transport, and secretion was dependent upon free concentrations of chlorothiazide in the perfusate. The maximum velocity of secretion (Vm = 85.6 micrograms/min) and Michaelis constant (Km = 2.1 micrograms/ml) of chlorothiazide was estimated using a nonlinear least-squares regression program. These results suggest that for compounds of low renal extraction, free drug concentrations are the driving force for carrier-mediated tubular secretion.