The effects of 1-methyl-5-thiotetrazole in a rat liver vitamin K-dependent carboxylase assay.
1-Methyl-5-thiotetrazole (NMTT), a metabolite of moxalactam (MoxamR), was studied for its potential inhibition of vitamin K-dependent carboxylation. The assay system utilized a detergent solubilized rat liver microsomal preparation. Vitamin K1H2 was artificially produced in situ by the NADH-dependent reduction of exogenous phylloquinone and the resultant carboxylation monitored by 14CO2 incorporation into a soluble peptide substrate. Warfarin, used as a reference inhibitor, gave results expected from the literature - 50% inhibition at a pharmacologically excessive level of 1.0 mM. Carboxylation was unaffected by 1.0 mM NMTT and was marginally (0-14%) diminished by 5.0 mM NMTT. Carboxylation was 25% diminished at 10.0 mM NMTT, a concentration far above that achieved in human testing of moxalactam. When NMTT was pre-incubated with the liver microsomal carboxylase enzyme preparation, 10.0 mM NMTT again caused merely a 25% diminution of carboxylation in the assay. These results do not support a role for NMTT as an inhibition of Vitamin K-dependent carboxylation which would produce pharmacological side effects during moxalactam therapy. During these studies it was found that dramatic consumption of NADH occurs in the presence of liver microsomal preparations (independent of vitamin K and of NMTT) and that NMTT effects on these processes may explain the small carboxylation diminution observed at 10.0 nM NMTT in the carboxylase assay.
D000818:Animals; D001393:Azoles / Q000494:pharmacology*; D019735:Carbon-Carbon Ligases; D006470:Hemorrhage / Q000139:chemically induced; D006801:Humans; D066298:In Vitro Techniques; D008025:Ligases / Q000037:antagonists & inhibitors*; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology; D009070:Moxalactam / Q000009:adverse effects; D051381:Rats; D011919:Rats, Inbred Strains; D013777:Tetrazoles / Q000494:pharmacology*; D014859:Warfarin / Q000494:pharmacology
D001393:Azoles; D013777:Tetrazoles; C038625:1-N-methyl-5-thiotetrazole; D014859:Warfarin; D008025:Ligases; D019735:Carbon-Carbon Ligases; C020660:glutamyl carboxylase; D009070:Moxalactam