Metabolism of 4,4'-methylene-bis-2-chloroaniline (MOCA) by rats in vivo and formation of N-hydroxy MOCA by rat and human liver microsomes.
The metabolism of 4,4'-methylene-bis-2-chloroaniline (MOCA) was investigated because it is an animal carcinogen to which humans have been exposed. In CD rats, where MOCA is a hepatocarcinogen, less than or equal to 0.2% of an oral dose of [14C]MOCA was recovered unchanged in the urine; enzymatic hydrolysis and extraction of urinary radioactivity indicated the presence of glucuronide and sulfate conjugates. In rat bile, the predominant metabolite was N-glucuronyl MOCA. Liver microsomes from male CD rats or human males (surgical specimens) were incubated in vitro with [14C] MOCA. Metabolite formation, which was dependent upon reduced pyridine nucleotides and intact microsomes, was quantitated by TLC and HPLC using appropriate chemically synthesized standards. N-Hydroxylation of MOCA occurred at a rate of 335 +/- 119 pmol/min/mg rat microsomal protein (n = 3) versus 230 or 765 (n = 2) with microsomes from humans; the product was identified by isotopic dilution for both species. The rates of 5-hydroxy-MOCA (o-aminophenol) formation were 92 +/- 33 (rats) and 7, 35 (human); rates for the benzhydrol derivative were 82 +/- 12 (rats) and 60, 160 (human). In rats, all three rates were elevated 4- to 8-fold by pretreatment with phenobarbital, which also enhanced the formation of partially characterized polar derivatives that appeared to result from oxidation and cleavage at the methylene carbon. The latter pathway typically amounted to 50-100% of the 4,4'-diamino-3,3'-dichlorobenzhydrol value in control or pretreated animals. Thus, rats metabolize MOCA extensively and the pathways include N-hydroxlation, which is regarded as an obligatory step in metabolic activation of arylamines. The presence of MOCA N-hydroxylase in human liver supports the hypothesis that exposure of humans to MOCA entails a carcinogenic risk.
Morton
K C
KC
Department of Chemical Carcinogenesis, Michigan Cancer Foundation, Detroit 48201-1380.
Lee
M S
MS
Siedlik
P
P
Chapman
R
R
D000818:Animals; D001559:Benzhydryl Compounds / Q000378:metabolism*; D001646:Bile / Q000378:metabolism; D002273:Carcinogens / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D005260:Female; D005965:Glucuronates / Q000378:metabolism; D006801:Humans; D006900:Hydroxylation; D066298:In Vitro Techniques; D008297:Male; D008753:Methylenebis(chloroaniline) / Q000378:metabolism*; D008862:Microsomes, Liver / Q000378:metabolism*; D051381:Rats
D001559:Benzhydryl Compounds; D002273:Carcinogens; D005965:Glucuronates; D008753:Methylenebis(chloroaniline)