Benzoyl-CoA conjugate accumulation as an initiating event for male reprotoxic effects in the rat? Structure-activity analysis, species specificity, and in vivo relevance.

A number of para-substituted benzoic acids (p-BA) and chemicals metabolized to p-BA have been found to confer adverse effects in male rats on sperm viability, motility, and morphology. These effects are putatively associated with the metabolism of p-BA to toxic intermediates. We had shown that p-BA lead to accumulation of high levels of p-alkyl-benzoyl-CoA conjugates in plated primary rat hepatocytes. Here we further investigated the relevance of this metabolic pathway for the reprotoxic effects in rats and rabbits. We extended the structure-activity relationship to a set of 19 chemicals (nine reprotoxic and ten non-reprotoxic) and confirmed a very strong correlation between p-alkyl-benzoyl-CoA accumulation in rat hepatocytes and the toxic outcome. Species specificity was probed by comparing rat, rabbit and human hepatocytes, and p-benzoyl-CoA accumulation was found to be specific to the rat hepatocytes, not occurring in human hepatocytes. There was also very limited accumulation in hepatocytes from rabbits that are a non-responder species in in vivo studies. Tissues of rats treated with 3-(4-isopropylphenyl)-2-methylpropanal were analysed and p-isopropyl-benzoyl-CoA conjugates were detected in the liver and in the testes in animals at toxic doses indicating that the metabolism observed in vitro is relevant to the in vivo situation and the critical metabolite does also occur in the reproductive tissue. These multiple lines of evidence further support benzoyl-CoA accumulation as a key initiating event for a specific group of male reproductive toxicants, and indicate a species-specific effect in the rat.