Integrative Experimental And Computational Study on the Binding of Potential Modulators with Glucokinase and Trypanosoma cruzi Cysteine Synthase for Drug Discovery

Type 2 diabetes mellitus (T2DM) affects a large portion of the population today and presents a significant global health concern due to its prevalence. It is estimated to impact more than 450 million people worldwide, with an expected continual increase. T2DM is characterised by defective insulin secretion, causing adverse effects including polydipsia, dysuria, fatigue and blurred vision amongst others. Due to its crucial role in the catalysis of glucose into glucose-6-phosphate (G6P), glucokinase (GK) is being actively researched in relation to T2DM and diseases linked to it. GK was also the main focus of this project, and a series of experimental and computational experiments were performed to study its activity. The main aim was to crystallise GK with an array of potential inhibitors, confirming their binding and comparing the results with molecular docking experiments performed on GOLD. GOLD is a widely recognised molecular docking software specialising in the prediction of protein-ligand binding. Crystals of GK were not obtained, however, comparison of binding assays and molecular docking experiments yielded interesting results, highlighting the conformational complexity of the protein. In addition to the study of GK, a second potential drug target Trypanosoma cruzi cysteine synthase (TcCS) was studied in relation to Chagas disease, a neglected tropical disease (NTD). The current treatments for Chagas disease are causative of many adverse effects and are not always effective. Gaining more knowledge and identifying potential drugs for the protein may be very beneficial. Here, TcCS was successfully expressed, purified and extensive co-crystallisation experiments with selected compounds resulted in several protein crystals being obtained. Tested compounds were part of an ensemble of compounds created during a drug-discovery campaign, that included fragment-based experiments and investigated a selection of inhibitors from bacterial homologues. Binding assays with these compounds were performed in parallel to help support X-ray crystallography experiments. Overall, the results contributed to the drug discovery campaign and will be utilised in biological studies in the future.