High-Throughput Kinetic Analysis for Target-Directed Covalent Ligand Discovery

Angewandte Chemie
Gregory B CravenDavid J Mann

Abstract

Cysteine-reactive small molecules are used as chemical probes of biological systems and as medicines. Identifying high-quality covalent ligands requires comprehensive kinetic analysis to distinguish selective binders from pan-reactive compounds. Quantitative irreversible tethering (qIT), a general method for screening cysteine-reactive small molecules based upon the maximization of kinetic selectivity, is described. This method was applied prospectively to discover covalent fragments that target the clinically important cell cycle regulator Cdk2. Crystal structures of the inhibitor complexes validate the approach and guide further optimization. The power of this technique is highlighted by the identification of a Cdk2-selective allosteric (type IV) kinase inhibitor whose novel mode-of-action could be exploited therapeutically.

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Citations

May 18, 2020·Angewandte Chemie·Mingxing TengNathanael S Gray
Jul 14, 2020·Chembiochem : a European Journal of Chemical Biology·Gregory B CravenDavid J Mann
Apr 2, 2020·Journal of Hematology & Oncology·Sha-Sha ChengDik-Lung Ma
Dec 5, 2020·Future Medicinal Chemistry·Elena De Vita
Jan 15, 2019·European Journal of Medicinal Chemistry·Weiyan ChengQuancheng Kan
Sep 4, 2019·Bioorganic & Medicinal Chemistry Letters·Concepción Sánchez-MartínezAlfonso de Dios
Oct 8, 2019·European Journal of Medicinal Chemistry·Yunyuan HuangJian Wan
Feb 5, 2021·Science Advances·Fandi SutantoAlexander Dömling
May 16, 2021·Nature Communications·Mitsugu ArakiYasushi Okuno
May 19, 2021·The Journal of Organic Chemistry·Gregory B CravenAlan Armstrong
Jun 1, 2021·ACS Central Science·Vincent M CrowleyBenjamin F Cravatt
Jan 9, 2020·Journal of Medicinal Chemistry·Daniel A ErlansonPaul N Mortenson
May 8, 2019·Journal of the American Chemical Society·Efrat ResnickNir London
Aug 31, 2021·RSC Chemical Biology·Wenchao LuNathanael S Gray

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Methods Mentioned

BETA
NMR
Assay
pulldown

Software Mentioned

qIT

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