The use of human in vitro metabolic parameters to explore the risk assessment of hazardous compounds: the case of ethylene dibromide.
Ethylene dibromide (1,2-dibromoethane, EDB) is metabolized by two routes: a conjugative route catalyzed by glutathione S-transferases (GST) and an oxidative route catalyzed by cytochrome P450 (P450). The GST route is associated with carcinogenicity. An approach is presented to use human purified GST and P450 enzymes to explore the importance of these metabolic pathways for man in vivo. This strategy basically consists of four steps: (i) identification of the most important isoenzymes in vitro, (ii) scaling to rate per milligram cytosolic and microsomal protein, (iii) scaling to rate per gram liver, and (iv) incorporation of data in a physiologically based pharmacokinetic (PBPK) model. In the first step, several GST isoenzymes were shown to be active toward EDB and displayed pseudo-first-order kinetics, while the EDB oxidation was catalyzed by CYP2E1, 2A6, and 2B6, which all displayed saturable kinetics. In the second step, the predictions were in agreement with the measured activity in a batch of 21 human liver samples. In the third step, rat liver P450 and GST metabolism of EDB was predicted to be in the same range as human metabolism (expressed per gram). Interindividual differences in GST activity were modeled to determine "extreme cases." For the most active person, an approximately 1.5-fold increase of the amount of conjugative metabolites was predicted. Lastly, it was shown that the GST route, even at low concentrations, will always contribute significantly to total metabolism. In the fourth step, a PBPK model describing liver metabolism after inhalatory exposure to EDB was used. The saturation of the P450 route was predicted to occur faster in the rat than in man. The rat was predicted to have a higher turnover of EDB from both routes. Nevertheless, when all data are combined, it is crucial to recognize that the GST remains significantly active even at low EDB concentrations. The limitations and advantages of the presented strategy are discussed.
Ploemen
J P
JP
TNO Toxicology, Zeist, The Netherlands.
Wormhoudt
L W
LW
Haenen
G R
GR
Oudshoorn
M J
MJ
Commandeur
J N
JN
Vermeulen
N P
NP
de Waziers
I
I
Beaune
P H
PH
Watabe
T
T
van Bladeren
P J
PJ
D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000187:drug effects / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D015946:Ethylene Dibromide / Q000493:pharmacokinetics* / Q000633:toxicity*; D005982:Glutathione Transferase / Q000187:drug effects* / Q000378:metabolism; D015386:Hazardous Substances / Q000378:metabolism* / Q000633:toxicity*; D006801:Humans; D066298:In Vitro Techniques; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000187:drug effects* / Q000201:enzymology; D008862:Microsomes, Liver / Q000378:metabolism; D008954:Models, Biological; D010920:Placenta / Q000201:enzymology; D051381:Rats; D018570:Risk Assessment; D013045:Species Specificity
D015386:Hazardous Substances; D015946:Ethylene Dibromide; D003577:Cytochrome P-450 Enzyme System; D005982:Glutathione Transferase