Macromolecule-macromolecule interaction in drug distribution. III. Kinetic characterization of the uptake of fractionated [3H]heparin and the effect of plasma proteins in the perfused rat liver.

The concentration-dependent hepatic uptake of fractionated [3H]heparin, a macromolecular model drug, was kinetically characterized and the effect of plasma proteins, albumin and alpha-globulin, was evaluated in the perfused rat liver as part of an ongoing effort to elucidate the mechanism of interaction of macromolecular drugs with biological macromolecules and the role of this interaction in the drugs' distribution. In the absence of proteins, the uptake of fractionated [3H]heparin was saturable with the maximum uptake velocity (Vmax) of 7.6 pmol/min/g liver and the Michaelis constant (Km) of 32.2 nM, suggesting the involvement of a specialized transport. alpha-Globulin (8.0 mg/ml) reduced the uptake of fractionated [3H]heparin at lower heparin at lower heparin concentrations. However, albumin (40 mg/ml) did not affect the uptake of fractionated [3H]heparin, suggesting an insignificant interaction. Assuming that fractionated [3H]heparin bound to alpha-globulin cannot be uptaken and that the reduction in uptake was solely attributable to the saturable Scatchard-type binding of fractionated [3H]heparin to alpha-globulin, the dissociation constant (Kd) and the binding capacity (n) were estimated to be 2.1 nM and 0.002, respectively. In in vitro binding experiments by ultrafiltration, Kd and n were estimated as 168 nM and 0.5, respectively, for alpha-globulin and 1021 nM and 0.02, respectively, for albumin, suggesting lower affinity and higher capacity in vitro for each protein.(ABSTRACT TRUNCATED AT 250 WORDS)