A new metabolic pathway of morphine: in vivo and in vitro formation of morphinone and morphine-glutathione adduct in guinea pig

Abstract

The biliary excretion of morphine and its metabolites by guinea pigs after s.c. injection of morphine (25 mg/kg) was determined by high-performance liquid chromatography procedures. The amounts of morphinone (MO), morphinone-glutathione adduct (MO-GSH), morphine and morphine-3-glucuronide excreted over a 4-hr period were 1.27, 9.35, 1.13 and 7.54% of the administeral dose, respectively. In addition, trace amounts of morphine-GSH adduct and morphinone-cysteine adduct, derived from MO-GSH, were detected. Although MO-GSH formation from MO proceeded nonenzymatically with a relatively high rate, GSH S-transferases participated in the reaction, especially at lower GSH concentrations. Pretreatment of the animals with diethyl maleate and lithocholic acid-3-sulfate decreased the total biliary excretion of MO. On the other hand, pretreatment with naloxone increased the excretion of total MO. In vitro experiments using liver cytosolic preparations showed that lithocholic acid-3-sulfate inhibited the enzymatic formation of MO from morphine and of MO-GSH from MO. The effects of naloxone on MO formation in vitro were more complex. A reaction was stimulated at low and suppressed at higher concentrations. The results indicated that MO-GSH formation from morphine is inhibited by lithocholic acid-3-sulfate through actions on both morphine 6-dehydrogenase and GSH S-transferases. Low levels of naloxone were found to stimulate the morphine 6-dehydrogenase-mediated dehydrogenation of morphine to MO.