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In Vitro Model of the Gram-Negative Bacterial Cell Envelope for Investigation of Anti-Infective Permeation Kinetics

  • Florian Graef
    Florian Graef
    Department of Drug Delivery, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Center for Infection Research (HZI), Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
    Department of Pharmacy, Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
  • Robert Richter
    Robert Richter
    Department of Drug Delivery, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Center for Infection Research (HZI), Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
    Department of Pharmacy, Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
  • Verena Fetz
    Verena Fetz
    Department of Chemical Biology, HZI, German Center for Infection Research, Inhoffenstraße 7, 38124 Braunschweig, Germany
    School of Engineering and Science, Jacobs University Bremen, Campus Ring 1, 28759 Bremen, Germany
    More by Verena Fetz
  • Xabier Murgia
    Xabier Murgia
    Department of Drug Delivery, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Center for Infection Research (HZI), Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
    Department of Pharmacy, Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
  • Chiara De Rossi
    Chiara De Rossi
    Department of Drug Delivery, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Center for Infection Research (HZI), Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
  • Nicole Schneider-Daum
    Nicole Schneider-Daum
    Department of Drug Delivery, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Center for Infection Research (HZI), Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
  • Giuseppe Allegretta
    Giuseppe Allegretta
    Department of Drug Design and Optimization, HIPS, HZI, Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
  • Walid Elgaher
    Walid Elgaher
    Department of Drug Design and Optimization, HIPS, HZI, Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
  • Jörg Haupenthal
    Jörg Haupenthal
    Department of Drug Design and Optimization, HIPS, HZI, Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
  • Martin Empting
    Martin Empting
    Department of Pharmacy, Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
    Department of Drug Design and Optimization, HIPS, HZI, Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
  • Felix Beckmann
    Felix Beckmann
    Institute of Materials Research, Helmholtz-Zentrum Geesthacht, Max-Planck-Straße 1, 21502 Geesthacht, Germany
  • Mark Brönstrup
    Mark Brönstrup
    Department of Chemical Biology, HZI, German Center for Infection Research, Inhoffenstraße 7, 38124 Braunschweig, Germany
  • Rolf Hartmann
    Rolf Hartmann
    Department of Pharmacy, Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
    Department of Drug Design and Optimization, HIPS, HZI, Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
  • Sarah Gordon*
    Sarah Gordon
    Department of Drug Delivery, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Center for Infection Research (HZI), Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
    School of Pharmacy and Biomolecular Sciences, Liverpool John Moores University, James Parsons Building, Byrom Street, L3 3AF Liverpool, United Kingdom
    *E-mail: [email protected]
    More by Sarah Gordon
  • , and 
  • Claus-Michael Lehr*
    Claus-Michael Lehr
    Department of Drug Delivery, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Center for Infection Research (HZI), Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
    Department of Pharmacy, Saarland University, Campus Building E8 1, 66123 Saarbrücken, Germany
    *E-mail: [email protected]
Cite this: ACS Infect. Dis. 2018, 4, 8, 1188–1196
Publication Date (Web):May 11, 2018
https://doi.org/10.1021/acsinfecdis.7b00165
Copyright © 2018 American Chemical Society

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    Abstract

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    The cell envelope of Gram-negative bacteria is a formidable biological barrier, inhibiting the action of antibiotics by impeding their permeation into the intracellular environment. In-depth understanding of permeation through this barrier remains a challenge, despite its critical role in antibiotic activity. We therefore designed a divisible in vitro permeation model of the Gram-negative bacterial cell envelope, mimicking its three essential structural elements, the inner membrane and the periplasmic space as well as the outer membrane, on a Transwell setup. The model was characterized by contemporary imaging techniques and employed to generate reproducible quantitative and time-resolved permeation data for various fluorescent probes and anti-infective molecules of different structure and physicochemical properties. For a set of three fluorescent probes, the permeation through the overall membrane model was found to correlate with in bacterio permeation. Even more interestingly, for a set of six Pseudomonas quorum sensing inhibitors, such permeability data were found to be predictive for their corresponding in bacterio activities. Further exploration of the capabilities of the overall model yielded a correlation between the permeability of porin-independent antibiotics and published in bacterio accumulation data; a promising ability to provide structure-permeability information was also demonstrated. Such a model may therefore constitute a valuable tool for the development of novel anti-infective drugs.

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    The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acsinfecdis.7b00165.

    • Nebulization chamber characterization, OM model characterization, OM model transport experiments, robustness assessment of the PS model, structural assessment of the overall envelope model, and overall envelope model transport experiments (RNAP inhibitors) (PDF)

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    Cited By

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