title: Chiral separation of new sulfonamide derivatives and evaluation of their enantioselective affinity for human carbonic anhydrase II by microscale thermophoresis and surface plasmon resonance.
abstract: The aim of this study was to develop a method combining chiral separation and biophysical techniques to evaluate the enantioselective affinity of original sulfonamide derivatives towards their therapeutic target, the human carbonic anhydrase II (hACII). The first step consisted in the preparation of the enantiomers by chromatographic separation. The performances of HPLC and Supercritical Fluid Chromatography (SFC) were studied at the analytical scale by optimization of various experimental conditions using adsorbed polysaccharide chiral stationary phases (amylose AD-H and cellulose OD-H). Since SFC allowed obtaining higher enantioresolutions per time unit, it was selected for the semi-preparative scale and successfully used to isolate each enantiomer with a satisfactory enantiomeric purity (>98%). Secondly, microscale thermophoresis (MST) method and surface plasmon resonance (SPR) used as reference method were developed to measure potential enantioselective affinities of these enantiomers towards the hACII. The optimizations of both methods were performed using a reference compound, i.e. acetazolamide, which affinity for hCAII has previously been demonstrated. For all compounds, KDvalues obtained using MST and SPR were in good agreement, leading to similar affinity scales despite both approaches totally differ (labeling for MST versus immobilization of the protein for SPR). The equilibrium dissociation constants of our original compounds for the hCAII were in the range 100-1000nM and an enantioselectivity was observed using the MST and SPR methods for the diarylpyrazole 2. Finally, by comparing the MST and SPR techniques, MST appears especially adapted for further screening of a series of sulfonamide derivatives due to the lower time required to estimate a binding constant while consuming as little hCAII as SPR.
authors.affiliation: Univ. Lille, EA 7365, GRITA - Groupe de Recherche sur les formes Injectables et les Technologies Associées, F-59000, Lille, France. Electronic address: tiphaine.rogez@univ-rouen.fr.
authors.forename: Tiphaine
authors.initials: T
authors.lastname: Rogez-Florent
authors.affiliation: Univ. Lille, EA 7365, GRITA - Groupe de Recherche sur les formes Injectables et les Technologies Associées, F-59000, Lille, France; Univ. Lille, Plate-forme d'interactions moléculaires, F-59000, Lille, France. Electronic address: catherine.foulon@univ-lille2.fr.
authors.forename: Catherine
authors.initials: C
authors.lastname: Foulon
authors.affiliation: Univ. Lille, Plate-forme d'interactions moléculaires, F-59000, Lille, France; CHU Lille, Banque de tissus, F-59000, Lille, France. Electronic address: anne-sophie.drucbert@univ-lille2.fr.
authors.forename: Anne-Sophie
authors.initials: AS
authors.lastname: Drucbert
authors.affiliation: Univ. Lille, EA 7365, GRITA - Groupe de Recherche sur les formes Injectables et les Technologies Associées, F-59000, Lille, France. Electronic address: nadege.schifano@univ-lille2.fr.
authors.forename: Nadège
authors.initials: N
authors.lastname: Schifano
authors.affiliation: Univ. Lille, Institut de Chimie Pharmaceutique Albert Lespagnol (ICPAL), F-59000, Lille, France. Electronic address: perrine.six@univ-lille2.fr.
authors.forename: Perrine
authors.initials: P
authors.lastname: Six
authors.affiliation: Univ. Lille, Institut de Chimie Pharmaceutique Albert Lespagnol (ICPAL), F-59000, Lille, France. Electronic address: stephanie.devassine@univ-lille1.fr.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Devassine
authors.affiliation: Univ. Lille, Institut de Chimie Pharmaceutique Albert Lespagnol (ICPAL), F-59000, Lille, France. Electronic address: patrick.depreux@univ-lille2.fr.
authors.forename: Patrick
authors.initials: P
authors.lastname: Depreux
authors.affiliation: Univ. Lille, Plate-forme d'interactions moléculaires, F-59000, Lille, France; CHU Lille, Banque de tissus, F-59000, Lille, France. Electronic address: pierre-marie.danze@univ-lille2.fr.
authors.forename: Pierre-Marie
authors.initials: PM
authors.lastname: Danzé
authors.affiliation: Univ. Lille, EA 7365, GRITA - Groupe de Recherche sur les formes Injectables et les Technologies Associées, F-59000, Lille, France; Univ. Lille, Institut de Chimie Pharmaceutique Albert Lespagnol (ICPAL), F-59000, Lille, France. Electronic address: laurence.goossens@univ-lille2.fr.
authors.forename: Laurence
authors.initials: L
authors.lastname: Goossens
authors.affiliation: Univ. Lille, EA 7365, GRITA - Groupe de Recherche sur les formes Injectables et les Technologies Associées, F-59000, Lille, France. Electronic address: cecile.danel-2@univ-lille2.fr.
authors.forename: Cécile
authors.initials: C
authors.lastname: Danel
authors.affiliation: Univ. Lille, EA 7365, GRITA - Groupe de Recherche sur les formes Injectables et les Technologies Associées, F-59000, Lille, France. Electronic address: jean-francois.goossens@univ-lille2.fr.
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Goossens
mesh_terms: D000086:Acetazolamide / Q000737:chemistry; D000688:Amylose / Q000737:chemistry; D024402:Carbonic Anhydrase II / Q000737:chemistry*; D002482:Cellulose / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D025924:Chromatography, Supercritical Fluid / Q000379:methods; D006801:Humans; D011134:Polysaccharides / Q000737:chemistry; D013237:Stereoisomerism; D013449:Sulfonamides / Q000737:chemistry*; D020349:Surface Plasmon Resonance / Q000379:methods
chemical_list: D011134:Polysaccharides; D013449:Sulfonamides; D002482:Cellulose; D000688:Amylose; D024402:Carbonic Anhydrase II; D000086:Acetazolamide
keywords: Chiral supercritical fluid chromatography; Diarylpyrazole sulfonamides; Human carbonic anhydrase II; Microscale thermophoresis; Surface plasmon resonance