Stereoselective glutathione conjugation of the separate alpha-bromoisovalerylurea and alpha-bromoisovaleric acid enantiomers in the rat in vivo and in rat liver subcellular fractions.

Glutathione (GSH) conjugation of the separate alpha-bromoisovalerylurea (BIU) enantiomers was studied in the rat. Administration of (R)-BIU resulted in excretion of a single glutathione conjugate in bile (IU-S-G/I) and a single mercapturate in urine (IU-S-MA/B). The other enantiomer, (S)-BIU, was exclusively metabolized to the other diastereomeric conjugates, IU-S-G/II and IU-S-MA/A. Thus, the conjugation of BIU with glutathione was completely stereospecific. Both the GSH conjugate and mercapturate derived from (R)-BIU were excreted two to three times more rapidly than their diastereomeric (S)-BIU counterparts. The enantiomers did not influence each others metabolism as reflected by identical metabolite excretion rates when the BIU enantiomers were administered either separately or as the racemic mixture. A similar rate difference for GSH conjugation of the separate BIU enantiomers was observed in incubations with rat liver cytosol as source of GSH transferases, suggesting that the stereoselectivity in vivo was due to glutathione conjugation properly. Similar results were obtained with a rat liver microsomal fraction, indicating that microsomal GSH transferases are active towards BIU and have a similar stereoselectivity as the cytosolic enzymes. Comparison of the GSH conjugation of BIU with that of its analogue alpha-bromoisovaleric acid (BI, which lacks the amide-linked urea group) revealed an opposite stereoselectivity: while (R)-BIU was conjugated faster than (S)-BIU, the (R) enantiomer of the acid was conjugated more slowly than (S)-BI. The alpha-bromocarbonyl compounds BI and BIU present a new type of substrate for the GSH transferases and allow studies of these enzymes in intact organisms as well as investigations on the stereoselectivity of GSH conjugation.