Use of pyridazinediones for tuneable and reversible covalent cysteine modification applied to peptides, proteins and hydrogels

Journal article

Over the past decades, the development of protein modification through cysteines, due their low abundance and high nucleophilicity, has enabled many advances in medicinal, imaging or diagnostics fields. In particular, their modification is of high interest for proteomic purposes, to inhibit the function of some proteins, or for the incorporation of any payload into a protein. On one hand, the development of probes/drugs for the targeting of cysteines in vivo has been clouded by their side, cytotoxic, effects. To overcome this, tools for the reversible modification of cysteines have been investigated. However, only a limited number of reagents have been reported so far and most of them suffer from stability issues, a lack of modularity and/or poor tunability. In this work, a library of tuneable nonbromo pyridazinediones (nonBr PD), as Michael acceptors, has been developed for their ability to react reversibly with cysteines. A correlation between the electrophilicity of the PDs and the Michael-addition/retro-Michael reaction rates was found. Those rates were reproducible onto a range of different cysteines, and the use of PD was exemplified on proteins/materials. Further investigations to understand the reactivity of those reagents led to the development of novel, improved, disulfide rebridging reagents. On the other hand, the focus has been on the development of chemical strategies for the stable modification of cysteine residues, applied to the generation of novel therapeutics, such as Antibody-Drug Conjugates (ADC) or bispecific formats for the treatment of cancer. Dibromo pyridazinediones (diBr PDs) scaffold have been previously reported for the homogeneous modification of disulfide bonds. Building on the works developed with diBr PDs and in collaboration with Wageningen University of Research and Queen’s University of Belfast, two novel bispecific formats based on site-selective modification strategies were developed and one of them was successfully tested in cell-based bioassays as a T-cell engager bispecific

Authors: Rochet, LNC; Bahou, C; Wojciechowski, JP; Koutsopetras, I; Britton, P

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: Royal Society of Chemistry
• Journal: Chemical Science
• Volume: 14
• Issue: 47
• Pages: 13743-13754
• Publication date: 2023-12-06
• DOI: 10.1039/d3sc04976k
• EISSN: 2041-6539
• ISSN: 2041-6520
• Language: English
• Keywords: Self-healing hydrogels; Biophysics; Chemical modification; Supramolecular chemistry; Biology; Organic chemistry; Molecule; Covalent bond; Exemplification; Biochemistry; Cysteine; Materials science; Combinatorial chemistry; Enzyme; Polymer chemistry; Chemistry; Dynamic covalent chemistry