DRUG METABOLISM

摘要

The invention relates to the technical area of detoxification and metabolism of drugs as well as endogenous compounds. Particularly, the present invention relates to the physiological and pharmacological significance of glucuronidation and to the biotechnological production of glucuronides. Drug metabolites generated by UDP glycosyltransferases (UGTs) are needed for drug development and toxicity studies, especially in the context of metabolites in safety testing (MIST). Since chemical metabolite synthesis can be stereo chemically demanding and arduous, various biological approaches have been developed; however, no whole-cell biotransformation with recombinant microbes that expresses human UGTs could yet be achieved. In this invention human UDP glucose-6-dehydrogenase was expressed (UGDH) together with a UGT isoform in the fission yeast Schizosaccharomyces pombe and strains that catalyze the whole-cell glucuronidation of standard substrates at rates up to 150 µmol L"1 d"1 were generated. Moreover, two methods to obtain stable isotope-labelled glucuronide metabolites were established: The first makes use of a labelled aglycon, while the second employs 13C6-glucose as a metabolic precursor of isotope-labelled UDP-glucuronic acid (UDP-GA) and yields a sixfold labelled glucuronide. This system should lead to a significant facilitation in the production of both labelled and unlabeled drug glucuronides.