Direct Automated MALDI Mass Spectrometry Analysis of Cellular Transporter Function: Inhibition of OATP2B1 Uptake by 294 DrugsClick to copy article linkArticle link copied!
- Melissa S. UngerMelissa S. UngerCenter for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack Str. 10, 68163 Mannheim, GermanyInstitute of Medical Technology, Heidelberg University and Mannheim University of Applied Sciences, Theodor-Kutzer-Ufer 1-3, 68167 Mannheim, GermanyCellzome - a GlaxoSmithKline company, Meyerhofstr. 1, 69177 Heidelberg, GermanyMore by Melissa S. Unger
- Lena SchumacherLena SchumacherCenter for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack Str. 10, 68163 Mannheim, GermanyMore by Lena Schumacher
- Thomas EnzleinThomas EnzleinCenter for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack Str. 10, 68163 Mannheim, GermanyMore by Thomas Enzlein
- David WeigtDavid WeigtCenter for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack Str. 10, 68163 Mannheim, GermanyMore by David Weigt
- Maciej J. Zamek-GliszczynskiMaciej J. Zamek-GliszczynskiDrug Metabolism and Pharmacokinetics, GlaxoSmithKline, 1250 S Collegeville Road, Collegeville, Pennsylvania 19426, United StatesMore by Maciej J. Zamek-Gliszczynski
- Matthias SchwabMatthias SchwabDr. Margarete Fischer-Bosch-Institute for Clinical Pharmacology, Auerbachstr. 112, 70376 Stuttgart, GermanyCluster of Excellence iFIT (EXC2180) “Image-Guided and Functionally Instructed Tumor Therapies”, University of Tuebingen, 72076 Tuebingen, GermanyDepartments of Clinical Pharmacology, Pharmacy and Biochemistry, University of Tuebingen, Auf der Morgenstelle 8, 72076 Tübingen, GermanyMore by Matthias Schwab
- Anne T. NiesAnne T. NiesDr. Margarete Fischer-Bosch-Institute for Clinical Pharmacology, Auerbachstr. 112, 70376 Stuttgart, GermanyCluster of Excellence iFIT (EXC2180) “Image-Guided and Functionally Instructed Tumor Therapies”, University of Tuebingen, 72076 Tuebingen, GermanyMore by Anne T. Nies
- Gerard DrewesGerard DrewesCellzome - a GlaxoSmithKline company, Meyerhofstr. 1, 69177 Heidelberg, GermanyMore by Gerard Drewes
- Sandra SchulzSandra SchulzCenter for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack Str. 10, 68163 Mannheim, GermanyMore by Sandra Schulz
- Friedrich B. M. ReinhardFriedrich B. M. ReinhardCellzome - a GlaxoSmithKline company, Meyerhofstr. 1, 69177 Heidelberg, GermanyMore by Friedrich B. M. Reinhard
- Carsten Hopf*Carsten Hopf*Email: [email protected]; Phone: +49 (0)621/292-6802; Mailing Address: Prof. Dr. Carsten Hopf, Mannheim University of Applied Sciences, Paul-Wittsack Str. 10, 68163 Mannheim, Germany.Center for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack Str. 10, 68163 Mannheim, GermanyInstitute of Medical Technology, Heidelberg University and Mannheim University of Applied Sciences, Theodor-Kutzer-Ufer 1-3, 68167 Mannheim, GermanyMore by Carsten Hopf
Abstract
OATP2B1, a member of the solute carrier (SLC) transporter family, is an important mechanism of substrate drug uptake in the intestine and liver and therefore a determinant of clinical pharmacokinetics and site of drug–drug interactions. Other SLC transporters have emerged as pharmacology targets. Studies of SLC transporter uptake to-date relied on radioisotope- or fluorescence-labeled reagents or low-throughput quantification of unlabeled compounds in cell lysate. In this study, we developed a cell-based MALDI MS workflow for investigation of OATP2B1 cellular uptake by optimizing the substrate, matrix, matrix–analyte ratio, and matrix application and normalization method. This workflow was automated and applied to characterize substrate transport kinetics and to test 294 top-marketed drugs for OATP2B1 inhibition and quantify inhibitory potencies necessary for extrapolation of clinical drug–drug interaction potential. Intra-assay reproducibility of this MALDI MS method was high (CV < 10%), and results agreed well (83% overlap) with previously published radioisotope assay data. Our results indicate that fast and robust MALDI MS cellular assays could emerge as a high-throughput label-free alternative for direct assessment of drug transporter function in DDIs and toxicities as well as enable drug discovery for transporters as pharmacology targets.
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This article is cited by 9 publications.
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- Melissa S. Unger, Martina Blank, Thomas Enzlein, Carsten Hopf. Label-free cell assays to determine compound uptake or drug action using MALDI-TOF mass spectrometry. Nature Protocols 2021, 16
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, 5533-5558. https://doi.org/10.1038/s41596-021-00624-z
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