Determination of Ligand-Binding Affinity (Kd) Using Transverse Relaxation Rate (R2) in the Ligand-Observed 1H NMR Experiment and Applications to Fragment-Based Drug Discovery.
Date
2023-08-10ICR Author
Author
Liu, M
Mirza, A
McAndrew, PC
Thapaliya, A
Pierrat, OA
Stubbs, M
Hahner, T
Chessum, NEA
Innocenti, P
Caldwell, J
Cheeseman, MD
Bellenie, BR
van Montfort, RLM
Newton, GK
Burke, R
Collins, I
Hoelder, S
Type
Journal Article
Metadata
Show full item recordAbstract
High hit rates from initial ligand-observed NMR screening can make it challenging to prioritize which hits to follow up, especially in cases where there are no available crystal structures of these hits bound to the target proteins or other strategies to provide affinity ranking. Here, we report a reproducible, accurate, and versatile quantitative ligand-observed NMR assay, which can determine Kd values of fragments in the affinity range of low μM to low mM using transverse relaxation rate R2 as the observable parameter. In this study, we examined the theory and proposed a mathematical formulation to obtain Kd values using non-linear regression analysis. We designed an assay format with automated sample preparation and simplified data analysis. Using tool compounds, we explored the assay reproducibility, accuracy, and detection limits. Finally, we used this assay to triage fragment hits, yielded from fragment screening against the CRBN/DDB1 complex.
Collections
Subject
Ligands
Reproducibility of Results
Proton Magnetic Resonance Spectroscopy
Small Molecule Libraries
Drug Discovery
Protein Binding
Research team
Hit Discov Struct Design
Medicinal Chemistry 4
Medicinal Chemistry 3
Language
eng
Date accepted
2023-07-19
License start date
2023-08-10
Citation
Journal of Medicinal Chemistry, 2023, 66 (15), pp. 10617 - 10627
Publisher
AMER CHEMICAL SOC