Expression, purification and characterization of membrane proteins which are potential antifungal drug targets

"An estimated 1.5 million people die from invasive fungal infections (IFI) each year. As well as the increasing patient population (currently exacerbated by the influence of COVID-associated IFIs), the problem of increasing resistance to current therapies is worrying. The spread of antifungal resistance has been identified by the WHO and CDC as a serious threat to health. With limited treatment options there is an urgent need for new antifungal drugs acting via novel mechanisms of action. F2G Ltd have several antifungal programmes at different stages that are each believed to target membrane proteins. The phase 3 antifungal compound olorofim acts by inhibiting the mitochondrial membrane protein dihydroorotate dehydrogenase (DHODH). Additionally, a multi-spanning membrane protein has been identified as the target of a preclinical series of antifungals, and a third potential membrane protein target has been found, that requires active protein to progress.

Traditionally, membrane proteins have been difficult to work with so that preparation of recombinant versions of potential drug targets for inhibitor screening has not always been possible. However, this collaboration will combine Aston's expertise in membrane protein expression, particularly in yeast, and the use of innovative solubilisation techniques such as SMA lipid particles (SMALPs) to enable these membrane proteins to be purified in their native form, allowing their activity to be studied. The proposed target proteins will be expressed and purified, followed by the development of methods to study the interaction between the transmembrane protein and potential drugs at the molecular level. Assay development of novel membrane protein targets will allow screening for inhibitors that will drive the identification of starting points for the discovery of new drugs."