Reproducible and time-dependent modification of serum protein binding in Wistar Kyoto rats.

The theoretical basis of the influence of (alterations in) plasma protein binding on pharmacokinetics (PK) is well-established. In contrast, the impact of protein binding on pharmacodynamics has not been examined in a systematic manner. Here we present an experimental approach to modify serum protein levels and binding in the rat, in a robust, reproducible, and time-dependent manner.Male Wistar Kyoto rats were divided into three different groups. The control group (n=4) did not receive treatment. In the cannulation(-) group (n=6) the rats were instrumented with three permanent blood cannulas. The rats in the cannulation(+) group received in addition to the cannulation a subcutaneous injection of turpentine oil of 100 microl/100 g bodyweight. The effects were characterized in terms of 1) the time course of serum levels of albumin and alpha(1)-acid glycoprotein (AGP), and 2) the effect on the ex vivo serum protein binding of S(-)-propranolol.In control rats the AGP serum concentration was stable at a value of 169+/-16 microg/ml. In the cannulation(-) group a maximum ten- to fifteen-fold increase in serum AGP concentration was observed at 48 h post surgery, followed by a gradual return back to baseline within 1 week. In the cannulation(+) group a similar concentration-time profile for AGP was found, but without a complete return to baseline within 1 week and with a much higher variability. Ex vivo, an increase in AGP serum concentration from 55 to 675 microg/ml resulted in a profound decrease in the free fraction of S(-)-propranolol from 14+/-0.6 to 1.9+/-0.3%.In conclusion, through cannulation alone the serum protein levels and binding were modified in a robust, reproducible and time-dependent manner. Therefore this experimental approach is suitable for the investigation of the influence of protein binding on both pharmacokinetics and pharmacodynamics.