Vol. 30, Issue 3, 295-300, March 2002

N-Glucuronidation of Some 4-Arylalkyl-1H-Imidazoles by Rat, Dog, and Human Liver Microsomes

Sanna Kaivosaari, Jarmo S. Salonen, and Jyrki Taskinen

Department of Pharmacy, Division of Pharmaceutical Chemistry, University of Helsinki, Finland (S.K., J.T.); and Orion Pharma, Preclinical and Clinical Research and Development, Turku, Finland (J.S.S.)

N-Glucuronidation in vitro of six 4-arylalkyl-1H-imidazoles (both enantiomers of medetomidine, detomidine, atipamezole, and two other closely related compounds) by rat, dog, and human liver microsomes and by four expressed human UDP-glucuronosyltransferase isoenzymes was studied. Human liver microsomes formed N-glucuronides of 4-arylalkyl-1H-imidazoles with high activity, with apparent V_max values ranging from 0.59 to 1.89 nmol/min/mg of protein. In comparison, apparent V_max values for two model compounds forming the N-glucuronides 4-aminobiphenyl and amitriptyline were 5.07 and 0.56 nmol/min/mg of protein, respectively. Atipamezole showed an exceptionally low apparent K_m value of 4.0 µM and a high specificity constant (V_max/K_m) of 256 compared with 4-aminobiphenyl (K_m, 265 µM; V_max/K_m, 19) and amitriptyline (K_m, 728 µM; V_max/K_m, 0.8). N-Glucuronidation of medetomidine was highly enantioselective in human liver microsomes; levomedetomidine exhibited a 60-fold V_max/K_m value compared with dexmedetomidine. Furthermore, two isomeric imidazole N-glucuronides were formed from dexmedetomidine, but only one was formed from levomedetomidine. Dog liver microsomes formed N-glucuronides of 4-arylalkyl-1H-imidazoles at a low rate and affinity, with apparent V_max values ranging from 0.29 to 0.73 nmol/min/mg of protein and apparent K_m values from 279 to 1640 µM. Rat liver microsomes glucuronidated these compounds at a barely detectable rate. Four expressed human UDP-glucuronosyltransferase isoenzymes (UGT1A3, UGT1A4, UGT1A6, and UGT1A9) were studied for 4-arylalkyl-1H-imidazole-conjugating activity. Only UGT1A4 glucuronidated these compounds at an activity of about 5% of that measured for 4-aminobiphenyl. The observed activity of UGT1A4 does not explain the high efficiency of glucuronidation of 4-arylalkyl-1H-imidazoles in human liver microsomes.