Léa N. C. Rochet, Calise Bahou, Jonathan P. Wojciechowski, Ilias Koutsopetras, Phyllida Britton, Richard J. Spears, ORCID logo a Ioanna A. Thanasi, Baihao Shao, Lisha Zhong, Dejan-Krešimir Bučar, Abil E. Aliev, Michael J. Porter, Molly M. Stevens, James R. Baker, and Vijay Chudasama
Chem. Sci. 2023
https://doi.org/10.1039/D3SC04976K
Reversible cysteine modification has been found to be a useful tool for a plethora of applications such as selective enzymatic inhibition, activity-based protein profiling and/or cargo release from a protein or a material. However, only a limited number of reagents display reliable dynamic/reversible thiol modification and, in most cases, many of these reagents suffer from issues of stability, a lack of modularity and/or poor rate tunability. In this work, we demonstrate the potential of pyridazinediones as novel reversible and tuneable covalent cysteine modifiers. We show that the electrophilicity of pyridazinediones correlates to the rates of the Michael addition and retro-Michael deconjugation reactions, demonstrating that pyridazinediones provide an enticing platform for readily tuneable and reversible thiol addition/release. We explore the regioselectivity of the novel reaction and unveil the reason for the fundamental increased reactivity of aryl bearing pyridazinediones by using DFT calculations and corroborating findings with SCXRD. We also applied this fundamental discovery to making more rapid disulfide rebridging agents in related work. We finally provide the groundwork for potential applications in various areas with exemplification using readily functionalised “clickable” pyridazinediones on clinically relevant cysteine and disulfide conjugated proteins, as well as on a hydrogel material.
