The use of bromopyridazinedione derivatives in chemical biology

Abstract

Tools that facilitate the chemical modification of peptides and proteins are gaining an increasing amount of interest across many avenues of chemical biology as they enable a plethora of therapeutic,...

Modular chemical construction of IgG-like mono- and bispecific synthetic antibodies (SynAbs)

“…As demonstrated before in our group, 32,34 a crucial advantage of the Br2PD-based method is the selective re-bridging of solvent-accessible disulfides in proteins. This leads to the controlled introduction of only one PD per Fab fragment, which presents only one accessible interchain disulfide; and two PDs on the Fc moiety, which has two accessible disulfides located in its hinge region (in case of an IgG1 parent isoform such as rituximab).…”

“…17,24,27,28 A subset of cysteinereactive modalities, disulfide re-bridging reagents, rely on the reduction of accessible disulfide bridges followed by their covalent reconnection via a small molecule -indeed these strategies have been used for the generation of protein-protein conjugates. 29,30 For this purpose, the Chudasama and Baker groups developed the pyridazinedione (PD) scaffold, [31][32][33][34] a chemical platform bearing: 1) two leaving groups across the double bond (generally bromine atoms) capable of reacting with the two liberated sulfhydryl groups generated via disulfide reduction, allowing for the covalent re-bridging of the disulfide; 2) up to two chemical handles for orthogonal click reactions, allowing selective dual-modification of the protein.…”

Modular Chemical Construction of IgG-like Mono- and Bispecific Synthetic Antibodies (SynAbs)

“…As demonstrated before in our group, , a crucial advantage of the Br 2 PD-based method is the selective rebridging of solvent-accessible disulfides in proteins. This leads to the controlled introduction of only one PD per Fab fragment, which presents only one accessible interchain disulfide; and two PDs on the Fc moiety, which has two accessible disulfides located in its hinge region (in case of an IgG1 parent isoform such as rituximab).…”

Enabling the formation of native mAb, Fab′ and Fc-conjugates using a bis-disulfide bridging reagent to achieve tunable payload-to-antibody ratios (PARs)

“…More recently, disulde bridging reagents have been developed, relying on the reduction of accessible disulde bridges (four in a IgG1 isoform) and their subsequent covalent reconnection via a small molecule. [11][12][13][14] Homogenous ADCs with controlled DAR could be generated through these chemical procedures. These methods, as well as enzymatic modication of canonical amino acids or N-glycan region, 15 can be directly applied to native antibodies.…”

“…To address the above challenges, and building on previous work in our lab, 13,14,27 we report a disulde re-bridging method that exploits the use of a novel trifunctional dual disulde rebridging linker (see Fig. 1).…”