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Multiparameter Kinetic Analysis for Covalent Fragment Optimization by Using Quantitative Irreversible Tethering (qIT)
Author(s) -
Craven Gregory B.,
Affron Dominic P.,
Kösel Teresa,
Wong Tsz Lam M.,
Jukes Zoë H.,
Liu ChunTing,
Morgan Rhodri M. L.,
Armstrong Alan,
Mann David J.
Publication year - 2020
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.202000457
Subject(s) - covalent bond , fragment (logic) , chemistry , cysteine , tethering , combinatorial chemistry , electrophile , stereochemistry , computer science , biochemistry , enzyme , organic chemistry , catalysis , programming language , operating system
Abstract Chemical probes that covalently modify cysteine residues in a protein‐specific manner are valuable tools for biological investigations. Covalent fragments are increasingly implemented as probe starting points, but the complex relationship between fragment structure and binding kinetics makes covalent fragment optimization uniquely challenging. We describe a new technique in covalent probe discovery that enables data‐driven optimization of covalent fragment potency and selectivity. This platform extends beyond the existing repertoire of methods for identifying covalent fragment hits by facilitating rapid multiparameter kinetic analysis of covalent structure–activity relationships through the simultaneous determination of K i , k inact and intrinsic reactivity. By applying this approach to develop novel probes against electrophile‐sensitive kinases, we showcase the utility of the platform in hit identification and highlight how multiparameter kinetic analysis enabled a successful fragment‐merging strategy.

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