Development of Factor XIIa Inhibitors Assisted by Affinity Capillary Electrophoresis, A Fragment-based Lead Discovery Project

Coagulation factor XIIa (FXIIa) is a S1 serine protease implicated in coagulation, inflammation, and immunity. The potential indications of FXIIa inhibitors include artificial surface-induced thrombosis, hereditary angioedema, Alzheimer’s disease, and multiple sclerosis [1]. Our team previously developed 3-carboxamido-benzopyrans [2]. The compounds were anticoagulants and showed some selectivity for the contact phase pathway [2c]. To support the modulations of the 3-carboxamido coumarins and to develop new chemical scaffolds, we implemented a fragment-based lead discovery platform. Fragment-based lead discovery is a usual strategy to identify innovative lead compounds. This approach relies on the screening of small-size molecules (< 300 g/mol). These fragments have a weak affinity for the target but explore more efficiently the protein chemical space. This strategy requires the detection of weak binders (μM–mM range) and characterization of their binding site to succeed. NMR and X-ray crystallography are the conventional techniques to address this challenge. However, their implementation is difficult due to their large protein consumption and the cost of equipment. Therefore, we investigated affinity capillary electrophoresis as a promising microfluidic technology for fragment-based lead discovery projects.
This communication is devoted to the presentation of the affinity capillary electrophoresis methodologies adapted for fragment-based lead discovery [3,4] and the results gathered on FXIIa. Their impact on our coumarin series will also be discussed.
[1] C. Davoine, C. Bouckaert, M. Fillet , L. Pochet, Eur. J. Med. Chem, 208 (2020), 10.1016/j.ejmech.2020.112753
[2] a) S. Robert, C. Bertolla, B. Masereel, J.M. Dogné, L. Pochet, Journal of Medicinal Chemistry, 51 (2008) 3077-3080; b) C. Bouckaert, S. Serra, G. Rondelet, E. Dolušić, J. Wouters, J.M. Dogné, R. Frédérick, L. Pochet, Eur J of Med Chem, 110 (2016) 181-194; c) C. Bouckaert , S. Zhu, J. W. P. Govers-Riemslag, M. Depoorter, S. L. Diamond, L. Pochet, Thromb Res, 157 (2017) 126-133.
[3] C. Davoine, M. Fillet, L. Pochet, Talanta, 226 (2021), 10.1016/j.talanta.2021.122163
[4] C. Davoine, A. Pardo, L. Pochet, M. Fillet, Analytical Chemistry, 93 (2021), 10.1021/acs.analchem.1c03611