Modern drug development is an astronomically expensive and time consuming undertaking. Because of this, studying the pharmacokinetic properties of drugs in vitro has become an integral step early in the process of drug development, with the goal of preventing costly failures late in the process, and dangerous side effects. Artificial phospholipid bilayers known as droplet interface bilayers (DIBs) have the potential to be used for these pharmacokinetics assays, combining the low cost of cell-free assays with the ability to more closely mimic structures found in life than current cell-free in vitro techniques. Combined with the reproducibility, ease of use, and low reagent consumption found with microfluidic methods, disruptive new low cost techniques for assessing pharmacokinetics in drug development may be possible using DIBs as an artificial cell membrane model.
In this work, I establish the potential of DIBs to be used as a pharmacokinetics modelling platform, and advance the use of microfluidic methods for carrying out pharmacokinetics assays in drug discovery. I first developed a new microfluidic platform for the formation of DIBs, which sought to solve some of the shortcomings of current microfluidic methods for DIB formation (Chapter 2). This device is the first that can be used to form DIB networks from dissimilar droplets in parallel, without use of active controls, and with droplet contact gentle enough to enable use of biomimetic lipid mixtures. I examine for the first time the behaviour of phospholipids on microfluidic devices, and characterise the interaction that they have with a common material used to construct microfluidic devices (Chapter 3). Not only has this interaction never been studied before, but my unexpected findings indicate a new area requiring further study in order to advance the adoption of DIBs on microfluidic devices. In collaboration with my colleague Jaime Korner, I use my newly developed microfluidic platform to carry out an on-chip permeation assay for the first time using biomimetic lipid formulations and bespoke compartments modelled after the human intestine. We demonstrate that this on-chip assay has predictive accuracy greater than that of a current widely used cell-free technique (Chapter 4). Finally, I demonstrate that a DIB based microfluidic platform enables, and is critical for, characterising the effect of structural features such as membrane asymmetry on drug permeation. With this, I find measurable, previously unknown effects of membrane asymmetry on the absorption of the chemotherapy drug doxorubicin, highlighting a possible contributing factor to chemoresistance in some cancers (Chapter 5). I find, and demonstrate throughout the body of this work that microfluidic methods and DIBs can not only provide alternatives to current cell-free in vitro pharmacokinetics assays, but that they can exceed the performance of existing assays, and be used for entirely new ways of examining pharmacokinetics. Through building bespoke artificial cell membranes from the ground up, I hope to demonstrate herein the great potential of these powerful new cell-free methods. / Graduate / 2022-09-12
| Identifer | oai:union.ndltd.org:uvic.ca/oai:dspace.library.uvic.ca:1828/13401 |
| Date | 20 September 2021 |
| Creators | Stephenson, Elanna |
| Contributors | Elvira, Katherine S. |
| Source Sets | University of Victoria |
| Language | English, English |
| Detected Language | English |
| Type | Thesis |
| Format | application/pdf |
| Rights | Available to the World Wide Web |
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