EFFECT OF STEREOCHEMISTRY ON THE OXIDATIVE-METABOLISM OF THE CYCLOPHOSPHAMIDE METABOLITE ALDOPHOSPHAMIDE

Abstract

P-31 NMR and cell perfusion techniques were used to investigate the conversion of the individual enantiomers of aldophosphamide (AP) to carboxyphosphamide (CBP) as catalyzed by aldehyde dehydrogenase in human erythroleukemia K562 cells. R- and S-cyclophosphamides (CPs) were treated with ozone and hydrogen peroxide to yield R(p)- and S-p-cis-4-hydroperoxycyclophosphamides (R(p)- and S-p-cis-4-HO2-CP); reduction of each hydroperoxide gave the corresponding enantiomer of AP [along with its tautomer 4-hydroxycyclophosphamide (4-HO-CP)]. In separate experiments, K562 cells embedded in agarose gel threads were perfused at pH 7.4, 21 +/- 1 degrees, with solutions of 1.4 mM R(p)- and S-p-4-HO-CP/AP, both with and without added mesna (an acrolein scavenger). A comparison of the P-32 NMR spectral data derived from the experiments revealed little statistical difference (+/- 10-20% error limits) in the normalized intensities of the CBP peaks arising from the individual AP enantiomers [with added mesna, the ratio R(p)-CBP:S-p-CBP was 1.00:1.24 +/- 0.13 (average deviation); without mesna, the same ratio was 1.00:1.35]. Using conventional methods for evaluating the in vitro drug toxicities, CP-resistant L1210 cells were treated in separate experiments with R(p)- and S-p-cis-4-HO2-CP; there were no significant differences between the toxicities exhibited by the stereoisomers.