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Direct Automated MALDI Mass Spectrometry Analysis of Cellular Transporter Function: Inhibition of OATP2B1 Uptake by 294 Drugs

  • Melissa S. Unger
    Melissa S. Unger
    Center for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack Str. 10, 68163 Mannheim, Germany
    Institute of Medical Technology, Heidelberg University and Mannheim University of Applied Sciences, Theodor-Kutzer-Ufer 1-3, 68167 Mannheim, Germany
    Cellzome - a GlaxoSmithKline company, Meyerhofstr. 1, 69177 Heidelberg, Germany
  • Lena Schumacher
    Lena Schumacher
    Center for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack Str. 10, 68163 Mannheim, Germany
  • Thomas Enzlein
    Thomas Enzlein
    Center for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack Str. 10, 68163 Mannheim, Germany
  • David Weigt
    David Weigt
    Center for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack Str. 10, 68163 Mannheim, Germany
    More by David Weigt
  • Maciej J. Zamek-Gliszczynski
    Maciej J. Zamek-Gliszczynski
    Drug Metabolism and Pharmacokinetics, GlaxoSmithKline, 1250 S Collegeville Road, Collegeville, Pennsylvania 19426, United States
  • Matthias Schwab
    Matthias Schwab
    Dr. Margarete Fischer-Bosch-Institute for Clinical Pharmacology, Auerbachstr. 112, 70376 Stuttgart, Germany
    Cluster of Excellence iFIT (EXC2180) “Image-Guided and Functionally Instructed Tumor Therapies”, University of Tuebingen, 72076 Tuebingen, Germany
    Departments of Clinical Pharmacology, Pharmacy and Biochemistry, University of Tuebingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany
  • Anne T. Nies
    Anne T. Nies
    Dr. Margarete Fischer-Bosch-Institute for Clinical Pharmacology, Auerbachstr. 112, 70376 Stuttgart, Germany
    Cluster of Excellence iFIT (EXC2180) “Image-Guided and Functionally Instructed Tumor Therapies”, University of Tuebingen, 72076 Tuebingen, Germany
    More by Anne T. Nies
  • Gerard Drewes
    Gerard Drewes
    Cellzome - a GlaxoSmithKline company, Meyerhofstr. 1, 69177 Heidelberg, Germany
  • Sandra Schulz
    Sandra Schulz
    Center for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack Str. 10, 68163 Mannheim, Germany
  • Friedrich B. M. Reinhard
    Friedrich B. M. Reinhard
    Cellzome - a GlaxoSmithKline company, Meyerhofstr. 1, 69177 Heidelberg, Germany
  • , and 
  • Carsten Hopf*
    Carsten Hopf
    Center for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack Str. 10, 68163 Mannheim, Germany
    Institute of Medical Technology, Heidelberg University and Mannheim University of Applied Sciences, Theodor-Kutzer-Ufer 1-3, 68167 Mannheim, Germany
    *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.
    More by Carsten Hopf
Cite this: Anal. Chem. 2020, 92, 17, 11851–11859
Publication Date (Web):July 28, 2020
https://doi.org/10.1021/acs.analchem.0c02186
Copyright © 2020 American Chemical Society

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    Abstract

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    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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    The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acs.analchem.0c02186.

    • Experimental setup, optimization of matrix solvents for DHAP and Ph-CCA-NH2, evaluation of the best cell number per spot, evaluation of the matrix suppression effect, comparison of automated and nonautomated assays, relationship of E3S intensity and concentration, overlap of MALDI and Karlgren data, autofluorescence and quenching results (PDF)

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    Cited By

    This article is cited by 8 publications.

    1. Melanie A. Felmlee, Michael Ng, Annie Lee. Expression and Pharmaceutical Relevance of Intestinal Transporters. 2023, 339-360. https://doi.org/10.1002/9781119660699.ch18
    2. Sergei Dikler. Accelerating Drug Discovery with Ultrahigh‐Throughput MALDI‐TOF MS. 2023, 393-421. https://doi.org/10.1002/9781119678496.ch12
    3. Martin Winter, Roman P. Simon, Tim T. Häbe, Robert Ries, Yuting Wang, David Kvaskoff, Amaury Fernández-Montalván, Andreas H. Luippold, Frank H. Büttner, Wolfgang Reindl. Label-free high-throughput screening via acoustic ejection mass spectrometry put into practice. SLAS Discovery 2023, 28 (5) , 240-246. https://doi.org/10.1016/j.slasd.2023.04.001
    4. Maria Emilia Dueñas, Rachel E Peltier‐Heap, Melanie Leveridge, Roland S Annan, Frank H Büttner, Matthias Trost. Advances in high‐throughput mass spectrometry in drug discovery. EMBO Molecular Medicine 2023, 15 (1) https://doi.org/10.15252/emmm.202114850
    5. Martin Winter, Roman P. Simon, Yuting Wang, Tom Bretschneider, Margit Bauer, Aniket Magarkar, Wolfgang Reindl, Amaury Fernández-Montalván, Florian Montel, Frank H. Büttner. Differential analyte derivatization enables unbiased MALDI-TOF-based high-throughput screening: A proof-of-concept study for the discovery of catechol-o-methyltransferase inhibitors. SLAS Discovery 2022, 27 (5) , 287-297. https://doi.org/10.1016/j.slasd.2022.05.002
    6. 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 (12) , 5533-5558. https://doi.org/10.1038/s41596-021-00624-z
    7. Roman P. Simon, Tim T. Häbe, Robert Ries, Martin Winter, Yuting Wang, Amaury Fernández-Montalván, Daniel Bischoff, Frank Runge, Wolfgang Reindl, Andreas H. Luippold, Frank H. Büttner. Acoustic Ejection Mass Spectrometry: A Fully Automatable Technology for High-Throughput Screening in Drug Discovery. SLAS Discovery 2021, 26 (8) , 961-973. https://doi.org/10.1177/24725552211028135
    8. Carina RamalloGuevara, Dorothea Paulssen, Anna A. Popova, Carsten Hopf, Pavel A. Levkin. Fast Nanoliter‐Scale Cell Assays Using Droplet Microarray–Mass Spectrometry Imaging. Advanced Biology 2021, 5 (3) https://doi.org/10.1002/adbi.202000279

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