A population pharmacokinetic-pharmacodynamic analysis of repeated measures time-to-event pharmacodynamic responses: the antiemetic effect of ondansetron.

This paper presents and illustrates methodology for specifying, estimating, and evaluating a predictive model for repeated measures time-to-event responses. The illustrative example specifies a model of the antiemetic effect vs. concentration relationship for the 5-HT3 antagonist ondansetron in the human ipecac model for emesis. A key part of this model is a time-dependent log hazard function for emesis that is increased by ipecac administration and decreased by ondansetron concentration. The model is fit using an approximate maximum likelihood method. The data consist of the time free of emeses and, for those individuals with emetic episodes, the time(s) of the episode(s). Model evaluation is accomplished using residual plots adapted to time-to-event data and a "posterior predictive check" wherein observed data statistics are compared to those obtained from data simulated from the fitted model. The ondansetron concentration required to obtain a 50% reduction in the hazard of emesis is estimated to be 1.4 +/- 0.2 ng/ml, and the rate constant for elimination of ipecac-induced hazard is 1.5 +/- 0.2 hr-1.

Tolerance to nitroglycerin in rabbit aorta. Investigating the involvement of the mu isozyme of glutathione S-transferases.

We have proposed that glutathione S-transferases (GSTs), especially the mu isozyme, play a critical role in the metabolism of nitroglycerin (glyceryl trinitrate, GTN), leading to pharmacologic effects. Here we study this enzyme(s) during tolerance development in male New Zealand white rabbits. Each aorta was divided into two segments designated as GTN pretreated and buffer control. Tolerance was induced in rabbit aortic strips so assigned by incubation with GTN (0.22 mM). The activity of the mu isozyme and of total GSTs was determined in portions f each segment. In each rabbit aorta, the response to GTN (0.5 microM) was determined in GTN-pretreated and buffer-pretreated strips by measuring cyclic GMP levels (N = 7 pairs) and percent relaxation (N = 4 pairs). In GTN-pretreated strips, a significant decrease was observed in the activity of the mu isozyme of GST, while the total GST activity was unchanged as compared with control strips. The decrease in isozyme activity correlated very well with the decrease in response to GTN. Two rabbit aortae did not become tolerant, and the activity of the mu isozyme was also not affected. The levels of thiols were not affected by GTN pretreatment and aortae tolerant to GTN did not develop tolerance to S-nitroso acetylpenicillamine (SNAP), indicating that thiol depletion and guanylate cyclase desensitization probably play a minor role in tolerance development to GTN in our model. These studies suggest that tolerance to GTN in rabbit aorta in vitro is associated with a decrease in GST mu activity, which correlates well with the decrease in GTN response.

Correlation of the response to nitroglycerin in rabbit aorta with the activity of the mu class glutathione S-transferase.

The relationship between the activity of glutathione S-transferases (GSTs), especially the mu isozyme, and the production of responses to nitroglycerin (GTN) was investigated in rabbit aorta. GST mu isozyme activity was measured using trans-stilbene oxide (TSO) as a substrate. Each aorta was divided into four parts, two of which were frozen for enzymatic analyses while the remaining two were used to measure the effects of GTN (0.5 microM), i.e. the increase in cGMP levels and the corresponding relaxation. Thus, all three measures were obtained in each individual rabbit aorta. Eight different rabbits were studied. An excellent correlation was obtained between the rise in cGMP and the mu isozyme activity (r2 = 0.948). A good correlation was also obtained between TSO activity and the relaxation response to GTN. Total GST activity did not correlate well with either cGMP increases or percent relaxation. These observations indicate that the activity of the mu isozyme measured using TSO and not the total GST correlates with the responses to GTN in the in vitro rabbit aorta model.

Effect of ethacrynic acid, a glutathione-S-transferase inhibitor, on nitroglycerin-mediated cGMP elevation and vasorelaxation of rabbit aortic strips.

The effects of ethacrynic acid (ECA), an inhibitor of glutathione-S-transferase, on both the pharmacologic and biochemical responses of aortic tissue to nitroglycerin (GTN) were evaluated. Using the rabbit aortic strip model, relaxation responses to 0.6 microM GTN were measured with and without ECA (0.2 mM) pretreatment. These same strips were frozen, and the concentrations of cGMP in the strips were measured using a 3H-labeled radioimmunoassay. Both the relaxation response and the increase in cGMP upon GTN treatment were reduced significantly by pretreatment of the strips with ECA. A correlation was observed between the decreases in the pharmacodynamic and biochemical responses upon ECA pretreatment. cGMP levels in strips treated with sodium nitroprusside, which generates nitric oxide by mechanisms distinct from that for organic nitrates, were not decreased by ECA pretreatment. These observations suggest that the mechanism of GTN action involves a glutathione-S-transferase-mediated metabolic step for GTN and that the isozyme(s) involved in this activation process may be inhibited by ECA.