Determination of Ligand-Binding Affinity (K d) Using Transverse Relaxation Rate (R 2) in the Ligand-Observed 1H NMR Experiment and Applications to Fragment-Based Drug Discovery

被引：8

作者：

Liu, Manjuan ^{[1
]}

Mirza, Amin ^{[1
]}

McAndrew, P. Craig ^{[1
]}

Thapaliya, Arjun ^{[1
]}

Pierrat, Olivier A. ^{[1
]}

Stubbs, Mark ^{[1
]}

Hahner, Tamas ^{[1
]}

Chessum, Nicola E. A. ^{[1
]}

Innocenti, Paolo ^{[1
]}

Caldwell, John ^{[1
]}

Cheeseman, Matthew D. ^{[1
]}

Bellenie, Benjamin R. ^{[1
]}

van Montfort, Rob L. M. ^{[1
,2
]}

Newton, Gary K. ^{[1
]}

Burke, Rosemary ^{[1
]}

Collins, Ian ^{[1
]}

Hoelder, Swen ^{[1
]}

机构：

[1] Inst Canc Res, Ctr Canc Drug Discovery, London SM2 5NG, England

[2] Inst Canc Res, Div Struct Biol, London SM2 5NG, England

来源：

JOURNAL OF MEDICINAL CHEMISTRY | 2023年 / 66卷 / 15期

关键词：

PROTEIN-DEGRADATION; NMR; GUIDE;

D O I：

10.1021/acs.jmedchem.3c00758

中图分类号：

R914 [药物化学];

学科分类号：

100701 ;

摘要：

High hit rates from initial ligand-observed NMR screeningcan makeit challenging to prioritize which hits to follow up, especially incases where there are no available crystal structures of these hitsbound to the target proteins or other strategies to provide affinityranking. Here, we report a reproducible, accurate, and versatile quantitativeligand-observed NMR assay, which can determine K (d) values of fragments in the affinity range of low & mu;Mto low mM using transverse relaxation rate R (2) as the observable parameter. In this study, we examined thetheory and proposed a mathematical formulation to obtain K (d) values using non-linear regression analysis. We designedan assay format with automated sample preparation and simplified dataanalysis. Using tool compounds, we explored the assay reproducibility,accuracy, and detection limits. Finally, we used this assay to triagefragment hits, yielded from fragment screening against the CRBN/DDB1complex.

引用

页码：10617 / 10627

页数：11

共 2 条

[1] 19F NMR viewed through two different lenses: ligand-observed and protein-observed 19F NMR applications for fragment-based drug discovery
Buchholz, Caroline R.
Pomerantz, William C. K.
RSC CHEMICAL BIOLOGY, 2021, 2 (05): : 1312 - 1330
[2] Ligand binding mode determination in Fragment Based Drug Discovery using X-ray crystallographic data coupled with QM/MM based refinement: Further applications of XModeScore
Borbulevych, Oleg Y.
Westerhoff, Lance M.
ACTA CRYSTALLOGRAPHICA A-FOUNDATION AND ADVANCES, 2018, 74 : A148 - A148

← 1 →