[en] Fragment-based lead discovery is a usual strategy in drug discovery to identify innovative lead compounds. The success of this approach strongly relies on the capacity to detect weak binders and characterize their binding site. NMR and X-ray crystallography are the conventional technologies used to tackle this challenge. However, their large protein consumption and the cost of equipment reduce their accessibility. Here, an affinity capillary electrophoresis methodology was developed that enables the detection of mM binders, the determination of dissociation constants, and the characterization of the fragment binding site. On the basis of multiple equilibrium theory, dissociation constants in the μM-mM range were determined, and a new methodology is proposed to establish graphically if two fragments bind the same protein pocket. The applicability of this methodology was demonstrated experimentally on coagulation factor XIIa by evaluating pairs of fragments with expected behavior. This study reinforces the significance of using affinity capillary electrophoresis to gather valuable information for medicinal chemistry projects.
Research center :
CIRM - Centre Interdisciplinaire de Recherche sur le Médicament - ULiège Namur Research Institute for Life Sciences - NARILIS
Davoine, Clara ; Université de Liège / Université de Namur - ULiège / UNamur > Département de pharmacie > CIRM - Analyse des Médicaments / NARILIS - NAMEDIC
Pardo, Alissia ; Université de Liège - ULiège > Département de pharmacie > CIRM > ét. visiteur médecine
Pochet, Lionel; Université de Namur - UNamur > Département de pharmacie > NARILIS - NAMEDIC
Fillet, Marianne ; Université de Liège - ULiège > Département de pharmacie > CIRM - Analyse des médicaments
Language :
English
Title :
Fragment Hit Discovery and Binding Site Characterization by Indirect Affinity Capillary Electrophoresis: Application to Factor XIIa
Publication date :
09 November 2021
Journal title :
Analytical Chemistry
ISSN :
0003-2700
eISSN :
1520-6882
Publisher :
American Chemical Society, United States - District of Columbia
Volume :
93
Pages :
14802–14809
Peer reviewed :
Peer Reviewed verified by ORBi
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] ULiège - Université de Liège [BE] UNamur - Université de Namur [BE]
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