Stimulation mechanism of guinea pig liver-mediated reduction of naloxone by morphine.
The mechanism of the stimulatory effect of morphine on the reduction of naloxone has been elucidated using guinea pig liver naloxone reductase that is identical with morphine 6-dehydrogenase. The reaction products were quantitated by means of HPLC. When naloxone was incubated with the enzyme in the presence of NAD(P)H at pH 7.4 or pH optima, the production of 6 alpha-naloxol increased according to the added amount of morphine. The stimulation was predominant with NADH at pH 7.4. Under these conditions, the production of morphinone also increased in proportion to the amount of morphine. The enzymatic reduction of naloxone proceeded even if NAD(P)H was replaced by NAD(P)+ and morphine. At a fairly low concentration of NADH (0.01 mM), the enzyme produced 6 alpha-naloxol (0.3 mM), exceeding the stoichiometric amount in the presence of 16 mM morphine. Although the Vmax values for naloxone was increased by the addition of morphine, the Km values for naloxone remained unaltered. Besides other substrates for guinea pig liver morphine 6-dehydrogenase such as codeine, normorphine and ethylmorphine also enhanced the reduction of naloxone. From these results we concluded that the stimulation of guinea pig liver-mediated reduction of naloxone by morphine is caused by the acceleration of the redox of pyridine nucleotides conducted by the enzyme. These phenomena were further supported by the experiments with the liver cytosol. In addition, we confirmed that, in the guinea pig, the biliary excretion of the metabolites, naloxol and naloxol-3-glucuronide, increased after sc injection of naloxone with morphine.
Kumagai
Y
Y
Faculty of Pharmaceutical Sciences, Fukuoka University, Japan.
Todaka
T
T
Yamano
S
S
Toki
S
S
D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D000818:Animals; D001646:Bile / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D006168:Guinea Pigs; D066298:In Vitro Techniques; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D009020:Morphine / Q000494:pharmacology*; D009022:Morphine Derivatives / Q000378:metabolism; D009243:NAD / Q000378:metabolism; D009270:Naloxone / Q000378:metabolism*; D010084:Oxidation-Reduction
D009022:Morphine Derivatives; D009243:NAD; D009270:Naloxone; D009020:Morphine; D000429:Alcohol Oxidoreductases; C045221:morphine 6-dehydrogenase; C010616:normorphine