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dc.contributor.authorRosenberger, Lara
dc.contributor.authorVon Essen, Carolina
dc.contributor.authorKhutia, Anupam
dc.contributor.authorKühn, Clemens
dc.contributor.authorUrbahns, Klaus
dc.contributor.authorGeorgi, Katrin
dc.contributor.authorHartmann, Rolf W.
dc.contributor.authorBadolo, Lassina
dc.date.accessioned2020-12-02T15:28:47Z
dc.date.available2020-12-02T15:28:47Z
dc.date.issued2020-07-01
dc.identifier.citationDrug Metab Dispos. 2020 Jul;48(7):587-593. doi: 10.1124/dmd.120.091140. Epub 2020 May 20.en_US
dc.identifier.issn00909556
dc.identifier.pmid32434832
dc.identifier.doi10.1124/DMD.120.091140
dc.identifier.urihttp://hdl.handle.net/10033/622629
dc.description.abstractUnderstanding the metabolism of new drug candidates is important during drug discovery and development, as circulating metabolites may contribute to efficacy or cause safety issues. In the early phase of drug discovery, human in vitro systems are used to investigate human relevant metabolism. Though conventional techniques are limited in their ability to provide complete molecular structures of metabolites (liquid chromatography mass spectrometry) or require a larger amount of material not available from in vitro incubation (nuclear magnetic resonance), we here report for the first time the use of the crystalline sponge method to identify phase I and phase II metabolites generated from in vitro liver microsomes or S9 fractions. Gemfibrozil was used as a test compound. Metabolites generated from incubation with microsomes or S9 fractions, were fractionated using online fraction collection. After chromatographic purification and fractionation of the generated metabolites, single crystal X-ray diffraction of crystalline sponges was used to identify the structure of gemfibrozil metabolites. This technique allowed for complete structure elucidation of 5'-CH2OH gemfibrozil (M1), 4'-OH gemfibrozil (M2), 5'-COOH gemfibrozil (M3), and the acyl glucuronide of gemfibrozil, 1-O-β-glucuronide (M4), the first acyl glucuronide available in the Cambridge Crystallographic Data Centre. Our study shows that when optimal soaking is possible, crystalline sponges technology is a sensitive (nanogram amount) and fast (few days) method that can be applied early in drug discovery to identify the structure of pure metabolites from in vitro incubations. SIGNIFICANCE STATEMENT: Complete structure elucidation of human metabolites plays a critical role in early drug discovery. Low amounts of material (nanogram) are only available at this stage and insufficient for nuclear magnetic resonance analysis. The crystalline sponge method has the potential to close this gap, as demonstrated in this study.en_US
dc.language.isoenen_US
dc.publisherAmerican Society for Pharmacology and Experimental Therapeuticsen_US
dc.rightsAttribution 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.titleCrystalline sponges as a sensitive and fast method for metabolite identification: Application to gemfibrozil and its phase I and II metabolitesen_US
dc.typeArticleen_US
dc.identifier.eissn1521009X
dc.contributor.departmentHIPS, Helmholtz-Institut für Pharmazeutische Forschung Saarland, Universitätscampus E8.1 66123 Saarbrücken, Germany.en_US
dc.identifier.journalDrug Metabolism and Dispositionen_US
dc.identifier.eid2-s2.0-85086747286
dc.identifier.scopusidSCOPUS_ID:85086747286
dc.source.volume48
dc.source.issue7
dc.source.beginpage587
dc.source.endpage593
refterms.dateFOA2020-12-02T15:28:48Z
dc.source.journaltitleDrug Metabolism and Disposition


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