Direct Optofluidic Measurement of the Lipid Permeability of Fluoroquinolones

Jehangir Cama,Stefano Pagliara,Silvia Hernández-Ainsa,Michael Schaich,Ulrich F Keyser,Kareem Al Nahas

doi:10.1038/srep32824

Abstract

Quantifying drug permeability across lipid membranes is crucial for drug development. In addition, reduced membrane permeability is a leading cause of antibiotic resistance in bacteria, and hence there is a need for new technologies that can quantify antibiotic transport across biological membranes. We recently developed an optofluidic assay that directly determines the permeability coefficient of autofluorescent drug molecules across lipid membranes. Using ultraviolet fluorescence microscopy, we directly track drug accumulation in giant lipid vesicles as they traverse a microfluidic device while exposed to the drug. Importantly, our measurement does not require the knowledge of the octanol partition coefficient of the drug – we directly determine the permeability coefficient for the specific drug-lipid system. In this work, we report measurements on a range of fluoroquinolone antibiotics and find that their pH dependent lipid permeability can span over two orders of magnitude. We describe various technical improvements for our assay, and provide a new graphical user interface for data analysis to make the technology easier to use for the wider community.

Highlights

A reduction in cell permeability to antibiotics is known to be an important biochemical mechanism of drug resistance in bacteria[1,2]
In this paper we extended our optofluidic measurement technique to investigate differences in the permeability of four fluoroquinolones over a range of pH values
The results obtained are validated by considering the chemical structures of the molecules; the permeability coefficients increase with an increase in the lipophilicity of the molecules, as expected

Summary

Introduction

A reduction in cell permeability to antibiotics is known to be an important biochemical mechanism of drug resistance in bacteria[1,2]. The pH of the surrounding environment plays a major role on a drug’s ability to permeate a lipid membrane – at different pH values, the drug molecule can exist in different charge states, which changes its lipophilicity[3,4]. This is of particular importance while studying the passage of a drug capsule through the human digestive tract – the pH changes from highly acidic in the stomach (1.0–2.5) to more neutral values (6.5–7.5) in the intestine[5]. These methods typically involve immobilizing the vesicles on chip and flowing in the drug; drug uptake into the vesicles is visualised using fluorescent reporters

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Conclusion