About the Project
Small molecule therapeutics represent a major contributor to human health and wellbeing both in developed and developing economies. The demand for new drugs is higher than ever and molecular interventions will continue to play a central role in all healthcare systems. However, productivity in pharmaceutical companies continues to drop with failures in drug discovery processes contributing to the spiralling cost. There is therefore a need to develop simple, efficient and reproducible low-cost high-throughput in-vitro assays that are capable of predicting in-vivo efficacy, early on in the drug development process. Of particular interest is the need to screen drug-membrane interactions that are now widely recognised to contribute to drug efficacy. However, it should be noted that there is likely to be a wealth of other (related) applications for such a technology, including screening protein-lipid interactions, as cheap sensors, cell sorting, tuning lipid ratios for transfection efficiencies etc..
Although there is a swathe of approaches for making individual lipid bilayers (such as supported lipid bilayers and droplet interface bilayers) that can mimic cell membranes, and with which small molecules can interact, these need development and de-skilling in order to be applicable as useful drug-membrane interaction assays. This PhD project proposes to evaluate the suitability of each model lipid bilayer type for development into a low-cost high-throughput assay for drug-membrane screening, then followed by down-selection, design and manufacture. Prototype design and manufacture is expected to revolve around additive manufacturing (e.g. 3D printing) and laser cutting hence expertise in 3D CAD software would be beneficial for this project.
Once manufactured, drug-membrane interaction studies will be carried out. These will involve fluorescence (for labelled or auto-fluorescent drugs) and FRAP (fluorescence recovery after photobleaching) measurements, which can indirectly probe drug-lipid membrane interactions for non-labelled drugs. As an example, to validate the performance of the device(s) the degree of membrane partitioning of tetracaine, a local anaesthetic known to membrane partition in a composition-dependent manner, will be assessed for an SOPC:cholesterol lipid compositional array and compared to known values.
This project will be highly interdisciplinary, with elements of mesoscale manufacturing, physical chemistry and pharmaceutical science.
References
1. Baciu, M., Sebai, S.C, Ces, O., Mulet, X., Clarke, J.A., Shearman, G.C., Law, R.V., Templer, R.H., Plisson, C., Parker, C.A., Gee, A., Degradative transport of cationic amphiphilic drugs across phospholipid bilayers. Phil. Trans. R. Soc. A, 2006; 35(3): 498-501;
2. Casey, D.R., Sebai, S.C., Shearman, G.C., Ces, O., Law, R.V., Templer, R.H., Gee, A.D. Formulation affects the rate of membrane degradation catalyzed by cationic amphiphilic drugs. Ind. Eng. Chem. Res., 2008; 47(3): 650-655.
3. Masters, T., Eng, W., Weng, Z.L., Arasi, B., Gauthier, N., Viasnoff, V. Easy fabrication of thin membranes with through holes. Application to protein patterning. PLOS ONE, 2012; 7(8): e44261.
4. Nguyen, T., Conboy, J.C. High-throughput screening of drug-lipid membrane interactions via counter-propagating second-generation imaging. Anal. Chem., 2011; 83(15): 5979-5988.
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