Glycosidases (GHs) are enzymes responsible for the degradation of carbohydrates and play many roles in human health and pathophysiology. Often, abnormal levels of glycosidase activity are markedly... Show moreGlycosidases (GHs) are enzymes responsible for the degradation of carbohydrates and play many roles in human health and pathophysiology. Often, abnormal levels of glycosidase activity are markedly linked to human pathologies. Example of this is the overexpression of heparanase (HPSE) in several cancer tissues. To date, the biomedical relevance of HPSE mostly pertains cancer treatment. This dissertation reports on the design, synthesis and biochemical evaluation of covalent mechanism-based inhibitors for heparanase, as well as the development of broad-spectrum activity-based probes (ABPs) for retaining exo- and endo-β-D-glucuronidases alike, including HPSE. The design of the novel inhibitors is supported by computational simulations, and the inhibition and selectivity of the newly synthesised compounds towards HPSE is demonstrated in vitro with the use of the newly developed ABPs. The thesis further builds on the concept of ABP-based profiling of GHs and discusses the development of inhibitors and probes targeting retaining β-L-arabinofuranosidases, a group of non-canonical glycosidases predominantly expressed by microorganisms of the human gut microbiome (HGM). As retaining β-L-arabinofuranosidases possess a contentious enzymatic mechanism, the novel chemical tools developed and presented in this thesis were decisive for unraveling β-L-arabinofuranosidases’ mechanism, and might serve in the future as probing tools for studies of the HGM. Show less
