Mechanism-Based Design of Quinoline Potassium Acyltrifluoroborates (KATs) for Rapid Amide-Forming Ligations at Physiological pH

26 April 2021, Version 1
This content is a preprint and has not undergone peer review at the time of posting.

Abstract

Potassium acyltrifluoroborates (KATs) undergo chemoselective amide-forming ligations with hydroxylamines. Under aqueous, acidic conditions these ligations can proceed rapidly, with rate constants of ~20 M-1 s-1. The requirement for lower pH to obtain the fastest rates, however, limits their use with certain biomolecules and precludes in vivo applications. By mechanistic investigations into the KAT ligation, including kinetic studies, X-ray crystallography, and DFT calculations, we have identified a key role for a proton in accelerating the ligation. We applied this knowledge to the design and synthesis of 8-quinolyl acyltrifluoroborates, a new class of KATs that ligates with hydroxylamines at pH 7.4 with rate constants >4 M-1 s-1. We trace the enhanced rate at physiological pH to unexpectedly high basicity of the 8-quinoline-KATs, which leads to their protonation even under neutral conditions. This proton assists the formation of the key tetrahedral intermediate and activates the leaving groups on the hydroxylamine towards a concerted 1,2-BF3shift that leads to the amide product. We demonstrate that the fast ligations at pH 7.4 can be carried out with a protein substrate at micromolar concentrations.

Keywords

bioconjugation techniques
kinetics data
amide formation reaction
Acylboronates
ligation

Supplementary materials

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Title
Matthias Yi-Chung Quinoline KAT SUPPORT ChemRxiv SUBMIT
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