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To assess the stability of a cisplatin (CDDP) complex prepared with chondroitin sulfate A (CSA) relative to protein binding in the circulation and kidney, a trichloroacetic acid (TCA) precipitation method was developed to measure the protein-unbound species of CDDP and the CDDP-CSA complex in plasma and kidney homogenates. The total and unbound drug concentrations were determined up to 3 h following a 2 mg/kg bolus injection of CDDP or CDDP-CSA complex to rats. The stability against plasma binding was evaluated by a determination of the area under concentration-time curve from time 0 to infinite time (AUC(0-infinity)); the ratio of unbound drug AUC(0-infinity) to total drug AUC(0-infinity) was employed to estimate the availability of the unbound drug in the circulation. The results showed that a competitive reaction to platinum existed between plasma protein and the CDDP-CSA complex, but the complex accounted for more than 60% of the protein-unbound species for a dosage, compared to 30% obtained by an administration of uncomplexed CDDP. The tissue binding kinetics in kidney for CDDP and the CDDP-CSA complex was investigated by the use of homogenates. The binding rate constants of CDDP and CDDP-CSA in a kidney homogenate were 0.0040 min(-1) and 0.0014 min(-1), respectively. The results indicate that the CDDP-CSA complex could effectively retard the binding of CDDP to protein in the kidney. These data provide evidence that endogenous protein is able to compete for platinum from the CDDP-CSA complex, but the complex effectively retarded the protein binding reaction with CDDP in plasma and kidney as compared to native CDDP, which has the potential for reducing the accumulation of CDDP in plasma and kidney.

Stability of a Cisplatin-Chondroitin Sulfate A Complex in Plasma and Kidney in Terms of Protein Binding.