[en] Fragment-based drug discovery is a usual approach to identify innovative lead compounds. This strategy relies on the screening of small-size compounds (< 300 g/mol). These fragments have a weak affinity for the target but explore more efficiently the protein chemical space. For successful applications, this strategy requires the detection of weak binders (µM–mM range) and characterization of their binding site. NMR and X-ray crystallography can address this challenge and, in consequence, are commonly used in fragment-based drug discovery. However, their accessibility is limited due to their large protein consumption and the cost of equipment. Here, we present an affinity capillary electrophoresis methodology that detects mM binders, determines dissociation constants, and characterizes the fragment binding site. We use the multiple equilibrium theory to model the binding events occurring in the capillary. On the basis of the resulting mathematical models, we determine dissociation constants in the μM–mM range, and we propose an original methodology to establish graphically if two fragments bind the same protein pocket [1].
[1] C. Davoine, A. Pardo, L. Pochet, M. Fillet, Analytical Chemistry, 2021, 10.1021/acs.analchem.1c03611
Research center :
CIRM - Centre Interdisciplinaire de Recherche sur le Médicament - ULiège NAmur Research Institute for LIfe Sciences (NARILIS)
Davoine, Clara ; Université de Liège - ULiège > Unités de recherche interfacultaires > Centre Interdisciplinaire de Recherche sur le Médicament (CIRM) ; UNamur - Université de Namur [BE] > Département de Pharmacie > NAmur MEdicine & Drug Innovation Center
Pochet, Lionel; UNamur - Université de Namur [BE] > Département de Pharmacie > NAmur MEdicine & Drug Innovation Center
Fillet, Marianne ; Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments
Language :
English
Title :
Affinity Capillary Electrophoresis for Fragment-based Drug Discovery Projects: Hit Discovery and Binding Site Characterization
Publication date :
11 September 2022
Event name :
12th International Symposium on Drug Analysis and the 32nd International Symposium on Pharmaceutical and Biomedical Analysis (DA-PBA 2022)
Event place :
Mons, Belgium
Event date :
September 11-14th 2022
Audience :
International
Peer reviewed :
Peer reviewed
Name of the research project :
Development of new compounds targeting coagulation factor XIIa using innovative microfluidic assays in the context of fragment-based drug discovery
Funders :
F.R.S.-FNRS - Fonds de la Recherche Scientifique [BE] Fonds Léon Fredericq [BE]