Disposition of 4-Methylbenzoylglycine in Rat Isolated Perfused Kidney and Effects of Hippurates on Renal Mitochondrial Metabolism

Abstract

Hippurates tend to accumulate within proximal tubule cells during renal secretion. High intracellular concentrations can alter proximal tubular function or lead to tubular toxicity. In this study we examined the renal disposition of the hippurate 4-methylbenzoylglycine, a compound known for its high renal intrinsic clearance in-vivo. The effect of intracellular accumulation on mitochondrial respiration was also measured in-vitro and compared with that of the 2-methyl and 4-amino analogues. Experiments were performed with either 2.5% pluronic or a combination of 2.2% pluronic and 2% bovine serum albumin (BSA) as oncotic agents. Within the concentration range studied (1-200 microg mL(-1)) tubular secretion seemed to be a function of the amount of unbound drug in the perfusate. Renal excretion data were best fitted by a model in which a Michaelis-Menten term was used to describe active secretion. Parameters obtained after the analysis of renal excretion data were the maximum transport velocity (TM = 55+/-2 microg min(-1)) and the Michaelis-Menten constant for tubular transport (KT = 4.2+/-0.8 microg mL(-1)). The compound accumulated extensively in kidney tissue, ratios up to 600 times the perfusate concentration were reached. Accumulation could be explained by active tubular uptake and data were analysed best by a model similar to the model used to describe renal excretion. Calculated parameters were theoretical maximum capacity (RM =300+/-210 microg g(-1)) and affinity constant for renal accumulation (KA = 5.0+/-4.4 microg mL(-1)). The high intracellular concentrations of 4-methylbenzoylglycine had no effect on kidney function and mitochondrial oxygen consumption. The 2-methyl analogue reduced mitochondrial respiration slightly, but 4-aminobenzoylglycine (p-aminohippurate) caused a significant reduction. In conclusion, this study shows that renal accumulation of a hippurate is determined by the efficiency of its tubular secretion. Whether the high intracellular concentrations affect tubular cell functioning depends on the analogue involved.

Uremic Toxins Inhibit Transport by Breast Cancer Resistance Protein and Multidrug Resistance Protein 4 at Clinically Relevant Concentrations

“…Since MRP4 and BCRP are located at the apical membrane of proximal tubule cells, transport activity depends on the intracellular levels of substrates rather than substrate concentrations in the blood. Previously, Masereeuw et al demonstrated that methyl hippuric acids accumulate during secretory transport in the isolated perfused rat kidney [20], [21]. Furthermore, they showed that 2-methyl hippuric acid levels were 175-times higher in kidney tissue compared to the perfusate and 4-methyl hippuric acid concentrations were even 600-times higher.…”