Carbonyl reductase activity for acetohexamide in human erythrocytes.
Acetohexamide is an oral antidiabetic agent and is metabolized by the reductive conversion of the acetoxy group to a secondary alcohol metabolite. In vivo, many drugs are metabolized by reductase enzymes; however, the characteristics of the enzymes that reduce carbonyl compounds need to be clarified. We tested whether reductase activity for acetohexamide can be found in human erythrocytes. Enzyme activity was monitored by formation of hydroxyhexamide using HPLC methods. In human erythrocytes, reductase activity (6.10 +/- 1.20 nmol/min/g hemoglobin) (mean +/- SD) was indeed observed, when 0.5 mM acetohexamide was used as a substrate. KM values and Vmax at the physiologically important pH 7.4 were 0.70 +/- 0.13 mM and 9.19 +/- 0.88 nmol/min/g hemoglobin, respectively. Separation of protein by gel filtration gave one major peak fraction with reductase activity whose molecular weight was estimated to be 31,000. Known substrates of carbonyl reductase such as menadione, daunorubicin, and ethacrynic acid inhibited the acetohexamide reduction. The acetohexamide reductase in erythrocyte showed characteristics of carbonyl reductase. Furthermore, acetohexamide reductase activity in erythrocyte was approximately 30% activity of that of human liver (0.17 +/- 0.05 nmol/min/mg cytosolic protein). The pattern of inhibitors in human liver was essentially the same as that in erythrocytes. It is plausible that the activity in erythrocytes may predict the activity in the liver. It was concluded that carbonyl reductase in human erythrocyte plays an important role in acetohexamide metabolism.
Kishimoto
M
M
First Department of Internal Medicine, University of Osaka.
Kawamori
R
R
Kamada
T
T
Inaba
T
T
D000092:Acetohexamide / Q000097:blood*; D000328:Adult; D000429:Alcohol Oxidoreductases / Q000037:antagonists & inhibitors / Q000097:blood* / Q000378:metabolism; D000449:Aldehyde Reductase; D000074408:Aldo-Keto Reductases; D002478:Cells, Cultured; D003630:Daunorubicin / Q000494:pharmacology; D004912:Erythrocytes / Q000201:enzymology*; D004976:Ethacrynic Acid / Q000494:pharmacology; D006454:Hemoglobins / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008297:Male; D009249:NADP / Q000097:blood; D010084:Oxidation-Reduction; D014812:Vitamin K / Q000494:pharmacology
D006454:Hemoglobins; D014812:Vitamin K; D009249:NADP; D000429:Alcohol Oxidoreductases; D000074408:Aldo-Keto Reductases; C064152:acetohexamide reductase; D000449:Aldehyde Reductase; D004976:Ethacrynic Acid; D000092:Acetohexamide; D003630:Daunorubicin