Frédérick, RaphaëlHubeau, SarahSarahHubeau2025-07-012025-07-012025-06-0920252025-06-09https://hdl.handle.net/2078.2/43271Pancreatic cancer, and most specifically pancreatic ductal adenocarcinoma (PDAC), is a particularly aggressive disease, characterized by low survival rates and a high metastatic capacity. Recent results have shown that super-invasive cells, derived from PDAC, express high level of pyruvate kinase M1, suggesting that this enzyme plays a key role in the invasive and migratory behavior of the PDAC cells. However, no specific PKM1 inhibitor has been discovered and developed to date, unlike PKM2 which has been extensively studied in oncology. The identification of new PKM1 inhibitors is an interesting strategy that was investigated for our project. A Fragment-Based Drug Design approach was adopted for this work. After the production and purification of the human recombinant PKM1 enzyme, biophysical screening by nanoDSF and enzymatic screening via coupled PKM1-LH, LDH and PKM1-Luciferase assays were performed on three libraires of chemical fragments. The selected molecules were resynthesized by reductive amination and characterized by NMR spectroscope and HPLC-MS. Ten fragments were initially identified as potential inhibitors. However, after their resynthesis, an almost total loss of enzymatic activity was observed, suggesting artefacts or interferences related to the libraries used. Several hypotheses were considered to explain this discrepancy including the presence of metal impurities, degradation of the compounds and the effects of storage in solution. This study highlights the importance of rigorous validation of fragment screening processes. Although, most of the identified fragments could not be confirmed as true inhibitors, the case of compound F0249 stands out. This fragment exhibits selective inhibitory activity on PKM1 and is currently under crystallographic analysis, opening promising avenues for future investigations in the development of targeted anti-metastatic therapies.pancreatic cancerpyruvate kinase M1metastasesinhibitorstext::thesis::master thesis