@misc{PorajKobielskaAtzrodtHollaetal., author = {Poraj-Kobielska, Marzena and Atzrodt, Jens and Holla, Wolfgang and Sandvoss, Martin and Gr{\"o}be, Glenn and Scheibner, Katrin and Hofrichter, Martin}, title = {Preparation of labeled human drug metabolites and drug-drug interaction-probes with fungal peroxygenases}, series = {Journal of Labelled Compounds and Radiopharmaceuticals}, volume = {56}, journal = {Journal of Labelled Compounds and Radiopharmaceuticals}, number = {9-10}, issn = {1099-1344}, doi = {10.1002/jlcr.3103}, pages = {513 -- 519}, abstract = {Enzymatic conversion of a drug can be an efficient alternative for the preparation of a complex metabolite compared with a multi-step chemical synthesis approach. Limitations exist for chemical methods for direct oxygen incorporation into organic molecules often suffering from low yields and unspecific oxidation and also for alternative whole-cell biotransformation processes, which require specific fermentation know-how. Stable oxygen-transferring biocatalysts such as unspecific peroxygenases (UPOs) could be an alternative for the synthesis of human drug metabolites and related stable isotope-labeled analogues. This work shows that UPOs can be used in combination with hydrogen/deuterium exchange for an efficient one-step process for the preparation of 4'-OH-diclofenac-d6. The scope of the reaction was investigated by screening of different peroxygenase subtypes for the transformation of selected deuterium-labeled substrates such as phenacetin-d3 or lidocaine-d3. Experiments with diclofenac-d7 revealed that the deuterium-labeling does not affect the kinetic parameters. By using the latter substrate and H2 (18) O2 as cosubstrate, it was possible to prepare a doubly isotope-labeled metabolite (4'-(18) OH-diclofenac-d6). UPOs offer certain practical advantages compared with P450 enzyme systems in terms of stability and ease of handling. Given these advantages, future work will expand the existing 'monooxygenation toolbox' of different fungal peroxygenases that mimic P450 in vitro reactions.}, language = {en} } @misc{KiebistHollaHeidrichetal., author = {Kiebist, Jan and Holla, Wolfgang and Heidrich, Johannes and Poraj-Kobielska, Marzena and Sandvoss, Martin and Simonis, Reiner and Gr{\"o}be, Glenn and Atzrodt, Jens and Hofrichter, Martin and Scheibner, Katrin}, title = {One-pot synthesis of human metabolites of SAR548304 by fungal peroxygenases}, series = {Bioorganic \& Medicinal Chemistry}, volume = {23}, journal = {Bioorganic \& Medicinal Chemistry}, number = {15}, issn = {0968-0896}, doi = {10.1016/j.bmc.2015.06.035}, pages = {4324 -- 4332}, abstract = {Unspecific peroxygenases (UPOs, EC 1.11.2.1) have proved to be stable oxygen-transferring biocatalysts for H2O2-dependent transformation of pharmaceuticals. We have applied UPOs in a drug development program and consider the enzymatic approach in parallel to a conventional chemical synthesis of the human metabolites of the bile acid reabsorption inhibitor SAR548304. Chemical preparation of N,N-di-desmethyl metabolite was realized by a seven-step synthesis starting from a late precursor of SAR548304 and included among others palladium catalysis and laborious chromatographic purification with an overall yield of 27\%. The enzymatic approach revealed that the UPO of Marasmius rotula is particularly suitable for selective N-dealkylation of the drug and enabled us to prepare both human metabolites via one-pot conversion with an overall yield of 66\% N,N-di-desmethyl metabolite and 49\% of N-mono-desmethylated compound in two separated kinetic-controlled reactions.}, language = {en} }