Projects per year
Identification of Differential Protein Binding Affinities in an Atropisomeric Pharmaceutical Compound by Noncovalent Mass Spectrometry, Equilibrium Dialysis, and Nuclear Magnetic Resonance
Research output: Contribution to journal › Article (Academic Journal)
4 Citations (Scopus)
Atropisomerism of pharmaceutical compounds is a challenging area for drug discovery programs.1 Strategies for dealing with these compounds include raising the energy barrier to atropisomerisation in order to develop the drug as a single isomer,2 or reducing the barrier to rotation and developing a mixture of rapidly interconverting isomers.3 Commonly, however, the atropisomers will be differentiated in terms of their affinity for a given protein target, and it is therefore important to rapidly identify the most active component prior to further compound development. We present equilibrium dialysis and saturation transfer difference-NMR (STD-NMR) as techniques for assessing relative affinities of an atropisomeric mixture against anti-apoptotic protein targets, Bcl-2 and Bcl-xL. These techniques require no prior separation of the mixture of compounds and are therefore rapid and simple approaches. We also explore the use of non-covalent mass spectrometry for determining KD values of individual atropisomers separated from the equilibrium mixture, and compare the results to solution phase measurements. Results from equilibrium dialysis, STD-NMR and non-covalent mass spectrometry are all in excellent agreement, and provide complementary information on differential binding, amplification of the strongest binders and KD values.
Maple, H. J., Garlish, R. A., Whitcombe, I., Hold, A., Prosser, C. E., Ford, D., Mackenzie, H., Crosby, J., Porter, J., Taylor, R. J., Crump, M. P., & Crump, M. P. (2013). Identification of Differential Protein Binding Affinities in an Atropisomeric Pharmaceutical Compound by Noncovalent Mass Spectrometry, Equilibrium Dialysis, and Nuclear Magnetic Resonance. Analytical Chemistry, 85(12), 5958–5964. https://doi.org/10.1021/ac400760p