Carrier-mediated transport of valacyclovir (vacv), the L-valyl ester prodrug of acyclovir (acv), via the human peptide transporter (hPepT1) has been shown in Xenopus laevis oocytes and in cell lines such as Chinese hamster ovary (CHO) and Caco-2 transfected with the hPepT1 gene. However, significant differences in vacv uptake were observed in those models as extracellular pH varied. The purpose of this work was to characterize the interactions of various ionic species of vacv with the peptide transporter by overexpressing the transporter gene, hPepT1, in CHO cells. Based on the pK(a) values of vacv, it was determined that vacv exists as four different ionic species (di-cationic, cationic, neutral and anionic) with a predominance of cationic and neutral species at physiologically relevant pH conditions. Vacv uptake was shown to increase with increasing pH of the extracellular medium from 5.5 to 7.2. The uptake value was maximal at around pH 7.2 and did not vary for studies done at higher pH. Vacv uptake was concentration dependent and saturable at all pH conditions (5.5, 6.2, 6.8, 7.5 and 7.9) with apparent Michaelis-Menten constants, mean (S.D.), of 7.42(0.32), 6.64(1.20), 5.38(0.88), 2.69(0.23) and 2.23(0.33) mM, respectively. The current results demonstrate that the estimated affinities of the cationic and the neutral species of vacv with hPepT1 are significantly different (7.4 versus 1.2 mM, respectively). Given the axial and radial (microclimate) pH gradients known to exist in the intestine, the greater than six-fold difference in affinity constants suggests that intestinal pH fluctuations may significantly impact upon the variability of vacv uptake.
Copyright 2000 John Wiley & Sons, Ltd.