Rapid and robust cysteine bioconjugation with vinylheteroarenes

Seki, Hikaru; Walsh, Stephen J.; Bargh, Jonathan D; Parker, Jeremy S.; Carroll, Jason S.; Spring, David R.

doi:10.1039/d1sc02722k

Cited by 26 publications

(26 citation statements)

References 61 publications

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“…To verify that the obtained maleimide handles on the DNA are suitable for bioconjugation reactions with proteins and to compare with the bioconjugation of the standard SMCC maleimide-modified DNA (see Figure A), we performed a series of bioconjugation experiments with albumin. Albumin was chosen as the model protein, as it contains one free thiol available for selective modification using thiol-selective reagents . Furthermore, it is highly relevant, as conjugation of albumin is a tool for half-life extension of drugs and biomacromolecules. , For the experiments we prepared maleimide, MBM and SMCC modified DNA on a 20 nmol scale and purified the conjugates by reverse-phase HPLC.…”

Section: Resultsmentioning

confidence: 99%

“…Albumin was chosen as the model protein, as it contains one free thiol available for selective modification using thiol-selective reagents. 26 Furthermore, it is highly relevant, as conjugation of albumin is a tool for half-life extension of drugs and biomacromolecules. 27,28 For the experiments we prepared maleimide, MBM and SMCC modified DNA on a 20 nmol scale and purified the conjugates by reverse-phase HPLC.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

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Preparation of Maleimide-Modified Oligonucleotides from the Corresponding Amines Using N-Methoxycarbonylmaleimide

2022

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Oligonucleotide conjugates constitute a versatile tool for research and bioanalytical purposes. Often, such conjugates are prepared by reaction between a thiol on the protein with a maleimide-modified oligonucleotide. Unlike most other chemical handles the maleimide functionality cannot be introduced directly during the solid-phase oligonucleotide synthesis, and therefore the standard method to introduce the maleimide functionality is to react an amino-modified DNA with a heterobifunctional linker containing an activated ester and a maleimide. Here, we present an alternative method for preparation of maleimide and monobromomaleimide-modified oligonucleotides from the corresponding amine using N-methoxycarbonylmaleimide and N-methoxycarbonylbromomaleimide, respectively. In this method, no additional linker is attached to the oligonucleotide, as the maleimide functionality is formed directly on the existing amine. The maleimide can thereby be positioned close to the oligonucleotide, providing a high degree of control over the final construct. The reaction occurs in 30–60 min under alkaline conditions. Maleimide-modified oligonucleotides prepared in this manner were conjugated to bovine serum albumin, and the reaction shows comparable reactivity to the corresponding oligonucleotide modified using the 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (SMCC) linker.

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Section: Resultsmentioning

confidence: 99%

Section: ■ Results and Discussionmentioning

confidence: 99%

Preparation of Maleimide-Modified Oligonucleotides from the Corresponding Amines Using N-Methoxycarbonylmaleimide

2022

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“…A plethora of exciting reagents and technologies exist that can facilitate the selective modification of cysteine residues and disulfide bonds, each displaying a unique set of properties and associated advantages and disadvantages. [13][14][15][16][17][18][19][20][21] Amongst some of the most popular highly reactive cysteine-targeting technologies (e.g. maleimide and maleimide derivatives) issues can arise with regard to bioconjugate stability (i.e.…”

Section: Introductionmentioning

confidence: 99%

The use of bromopyridazinedione derivatives in chemical biology

Bahou¹,

Chudasama²

2022

Org. Biomol. Chem.

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“…The warhead moiety in these drugs is essential to control the toxicity of targeting or off-targeting induced by a covalent interaction. , Notwithstanding, most covalent therapeutics are limited to an α,β-unsaturated amide serving as an electrophilic warhead, and the modification site mainly involves cysteine or lysine residues in proteins. To overcome this limitation, many novel electrophilic warheads such as 2-sulfonylpyridine, bicyclo[1.1.0]butane (BCB) carboxylic amide, cyanamide, sulfonyl fluoride, oxaziridine, sulfur-triazole, sulfotetrafluorophenyl (STP) ester, α-chlorofluoroacetamide, alkynyl benzoxazine, vinylheteroarene, vinyl/alkynyl pyridine, and cyclopropene/cyclopropenone , that target tyrosine, methionine, lysine, and cysteine residues have been developed in recent years. These electrophiles have already been employed in activity-based protein profiling for the discovery of new druggable targets and development of novel covalent inhibitors with excellent pharmaceutical properties …”

Section: Introductionmentioning

confidence: 99%

Ynamide Electrophile for the Profiling of Ligandable Carboxyl Residues in Live Cells and the Development of New Covalent Inhibitors

Zhang

et al. 2022

J. Med. Chem.

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Covalent inhibitors with an electrophilic warhead have received considerable attention due to their remarkable pharmacological properties. However, the electrophilic warhead in covalent drugs is often an α, β-unsaturated amide, and the targets are mainly cysteine or lysine residues. Thus, the development of novel electrophiles that can target other amino acids is highly desirable. Ynamide, a useful and versatile building block, is commonly employed in the construction of various compounds in organic synthesis. The performance of this functional group in a proteome-wide environment has been studied here for the first time, and it has been shown that it can efficiently modify carboxyl residues in situ and in vitro. Upon incorporation of this ynamide warhead into the pharmacophores of kinase inhibitors, the resulting compound showed moderate inhibition against the EGFR L858R mutant but not against EGFR WT. This novel electrophilic group can be used in the development of new types of covalent inhibitors.

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Rapid and robust cysteine bioconjugation with vinylheteroarenes

Abstract: Vinylheteroarene linkers can chemoselectively modify cysteine residues in proteins and antibodies. These linkers give stable bioconjugates, and were used to synthesise efficacious antibody-drug conjugates.

Cited by 26 publications

References 61 publications

Preparation of Maleimide-Modified Oligonucleotides from the Corresponding Amines Using N-Methoxycarbonylmaleimide

Preparation of Maleimide-Modified Oligonucleotides from the Corresponding Amines Using N-Methoxycarbonylmaleimide

The use of bromopyridazinedione derivatives in chemical biology

Ynamide Electrophile for the Profiling of Ligandable Carboxyl Residues in Live Cells and the Development of New Covalent Inhibitors

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