Chiral separation of new sulfonamide derivatives & evaluation of enantioselective affinity for human carbonic anhydrase II by MST & SPR

 

Tiphaine Rogez-Florent, Catherine Foulon, Anne-Sophie Drucbert, Nadège Schifano, Perrine Six, Stéphanie Devassine, Patrick Depreux, Pierre-Marie Danzé, Laurence Goossens,c, Cécile Danel & Jean-Francois Goossens

Journal of Pharmaceutical and Biomedical Analysis
2017 vol: 137 pp: 113-122 doi:10.1016/j.jpba.2017.01.023

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, KD values 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 100–1000 nM 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.

View Publication

Topics: SPR, Aromatic compounds, Small molecules, Monolith – MicroScale Thermophoresis, MST, Proteins, Publications

 

 

Previous Article
Quantifying CBM carbohydrate interactions using Microscale Thermophoresis
Quantifying CBM carbohydrate interactions using Microscale Thermophoresis

Next Article
Colorimetric detection of cholic acid based on an aptamer adsorbed gold nanoprobe
Colorimetric detection of cholic acid based on an aptamer adsorbed gold nanoprobe

×

Sign up to receive
the latest NanoTemper news, product updates, event announcements and more

First Name
Last Name
Company Name
State
Province
State
Region
State
Canton
Agree to Privacy Policy*
*I have fully read, understood and agree to the Privacy Policy. I accept the storing and processing of my personal data by NanoTemper as described in the Privacy Policy.

By completing this form, you provide us consent to contact you with educational content, news and information about our products, services and events. You may unsubscribe at any time.
Thank you!
Error - something went wrong!