Chloropicrin: reactions with biological thiols and metabolism in mice

Chloropicrin (CCl3NO2) is a preplant soil fumigant as a fungicide, warning agent for other fumigants, and former was gas. Its mode of action is unknown but presumably related to its facile metabolic dechlorination on reaction with biological thiols. These interactions were studied with 14CCl3NO2, 14CHCl2NO2, 14CH2ClNO2, and 14CH3NO2, synthesized for this purpose. Reaction of 14CCl3NO2 with GSH yields the di- and monochloro derivatives, GSSG, and a small amount of nitrite. The initial adduct with GSH is transient, but more stable derivatives are formed at longer times or on catalysis by GSH S-transferase. Human Hb, examined as a model thiol protein, reacts with 1.5 mol equiv of 14CCl3NO2 at pH 7.4 to produce 53-67% 14CHCl2NO2 and 10-11% Hb dducts (radiochemical yields) which, on longer incubation, release 14CHCl2NO2, CHCl2NO2 and CH2ClNO2 react with the same site(s) in Hb but more slowly. The dechlorination reactions are also catalyzed by liver cytosol and microsomes with NADPH. Dechlorination and adduct formation observed in vitro also occur in vivo in ip-treated mice. Thus, 14CCl3NO2 quickly gives 14CHCl2NO2, and some 14CO2 is formed, but the primary urinary metabolites are polar and nonvolatile. The major metabolite from 14CCl3NO2 in urine is less polar than those from 14CHCl2NO2, 14CH2ClNO2, and 14CH3NO2 which, in turn, are different from the 14CH2O and H14CO2Na metabolites. Liver tissue is adducted in vivo by 14CCl3NO2 (43% radiocarbon unextractable with methanol at 1 h), but the total radiocarbon content decreases 4-fold between 1 and 48 h after treatment. Liver effects from a high CCl3NO2 dose are associated with 19% less GSH and 50% more absorbance from soluble hemoproteins than controls. The toxicity of CCl3NO2 is probably due to disruption of multiple targets by its cascade of dechlorination products.