Reduction with tris(2-carboxyethyl)phosphine (TCEP) enables the use of an S-sulphonate protecting group for thiol-mediated bioconjugation

Maret, Barbara; Regnier, Jérémi; Rossi, Jean‐Christophe; Garrelly, Laurent; Vial, Laurent; Pascal, Robert

doi:10.1039/c3ra47407k

Cited by 11 publications

(8 citation statements)

References 25 publications

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“…The 1-thio sugars have attracted increasing attention and interest as a building block in the synthesis of glyco-conjugates especially for the modification of proteins by an oligosaccharide . Reduction of the unprotected β-glucosyl Bunte salt with tris(2-carboxyethyl) phosphine hydrochloride (TCEP-HCl) afforded the corresponding 1-thioglucose in 70% yield (Figure a). It should be noted that we were able to prepare 1-thio sugars through a one-pot process via glycosyl Bunte salt intermediates, starting from unprotected sugars.…”

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confidence: 99%

Glycosyl Bunte Salts: A Class of Intermediates for Sugar Chemistry

Meguro

Noguchi

et al. 2017

Org. Lett.

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S-Glycosyl thiosulfates have been discovered as a new class of synthetic intermediates in sugar chemistry, named "glycosyl Bunte salts" after 19th-century German chemist, Hans Bunte. The synthesis was achieved by direct condensation of unprotected sugars and sodium thiosulfate using a formamidine-type dehydrating agent in water-acetonitrile mixed solvent. The application of glycosyl Bunte salts is demonstrated with transformation reactions into other glycosyl compounds such as a 1-thio sugar, a glycosyl disulfide, a 1,6-anhydro sugar, and an O-glycoside.

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confidence: 99%

Glycosyl Bunte Salts: A Class of Intermediates for Sugar Chemistry

Meguro

Noguchi

et al. 2017

Org. Lett.

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“…The results observed here are consistent with previous observations that TCEP indeed reacts with maleimide to generate a phosphorus-containing adduct; however, it is proposed here that this adduct is closer in structure to the neutral ylene rather than the phosphonium ylide proposed previously. [27][28][29] Scheme 2. Reaction of lysine derivatized maleimide (5) with TCEP (1) and THPP (2).…”

Section: Resultsmentioning

confidence: 99%

“…[18][19][20][21][22][23][24] Contrary to this however, a number of groups have reported decreased yields of conjugation when using maleimide in the presence of TCEP, suggesting possible reaction between the two reagents. [25][26][27][28] Recently, Sánchez et al have reported the observation of a phosphonium-ion adduct between TCEP and maleimide on the basis of mass spectrometry, though this adduct was not fully characterised. 28 In this study we demonstrate that the water soluble phosphine reducing agents TCEP and THPP both react rapidly with maleimides and phenyl vinyl sulfone under conditions typically employed for bioconjugation reactions.…”

Section: Introductionmentioning

confidence: 99%

“…In addition to the favorable physicochemical properties of TCEP and THPP, these reagents are also considered generally to provide methodological advantages over thiol based reducing agents, with reports in the literature that TCEP is unreactive toward maleimide and numerous examples of maleimide based conjugations being performed in the presence of TCEP. − Contrary to this, however, a number of groups have reported decreased yields of conjugation when using maleimide in the presence of TCEP, suggesting a possible reaction between the two reagents. − Recently, Sánchez et al have reported the observation of a phosphonium ion adduct between TCEP and maleimide on the basis of mass spectrometry, although this adduct was not fully characterized …”

Section: Introductionmentioning

confidence: 99%

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Characterization of Reactions between Water-Soluble Trialkylphosphines and Thiol Alkylating Reagents: Implications for Protein-Conjugation Reactions

Kantner

Watts

2016

Bioconjugate Chem.

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Water-soluble trialkylphosphines such as tris(carboxyethyl)phosphine (TCEP) and trishydroxypropyl phosphine (THPP) are effective agents for reducing disulfide bonds in proteins and are increasingly becoming the reagents of choice for bioconjugation strategies that modify cysteine (thiol containing) amino acids. These reducing agents are often considered as being chemically compatible with Michael acceptors such as maleimides and, as such, are often not removed prior to performing protein conjugation reactions. Here, we demonstrate the rapid and irreversible reaction of both TCEP and THPP with derivatives of the commonly employed thiol alkylating groups, maleimide and vinyl sulfone. Mechanistic investigations revealed distinct differences between the reactions of TCEP and THPP with maleimide, leading to the production of either nonproductive ylenes or succidimidyl derivatives, respectively. Importantly, we also demonstrate the incorporation of nonproductive ylenes formed between maleimide and TCEP into the Pneumococcal capsular polysaccharide Pn6b following strategies employed toward the production of conjugate vaccines.

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“…34 Many others have since then confirmed the early observations, showing that an excess of the reducing agent is detrimental to one-pot thiol chemistry. 31,32,41,42 Conversely, TCEP has been used successfully in one-pot thiol chemistry with other alkenes, including vinyl sulfone, 43 vinyl phenyl, 44 and acrylate 45 moieties, and not only as reducing agent but as a catalyst for the thiol−ene reaction. In our study, it however appeared that an excess amount of TCEP is also detrimental for thiol/vinyl sulfone conjugation.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Adequate Reducing Conditions Enable Conjugation of Oxidized Peptides to Polymers by One-Pot Thiol Click Chemistry

et al. 2018

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Thiol(-click) chemistry has been extensively investigated to conjugate (bio)molecules to polymers. Handling of cysteine-containing molecules may however be cumbersome, especially in the case of fast-oxidizing coiled-coil-forming peptides. In the present study, we investigated the practicality of a one-pot process to concomitantly reduce and conjugate an oxidized peptide to a polymer. Three thiol-based conjugation chemistries (vinyl sulfone (VS), maleimide, and pyridyldithiol) were assayed along with three reducing agents (tris(2-carboxyethyl)phosphine (TCEP), dithiothreitol, and β-mercaptoethanol). Seven out of the nine possible combinations significantly enhanced the conjugation yield, provided that an adequate concentration of reductant was used. Among them, the coincubation of an oxidized peptide with TCEP and a VS-modified polymer displayed the highest level of conjugation. Our results also provide insights into two topics that currently lack consensus: TCEP is stable in 10 mM phosphate buffered saline and it reacts with thiol-alkylating agents at submillimolar concentrations, and thus should be carefully used in order to avoid interference with thiol-based conjugation reactions.

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Reduction with tris(2-carboxyethyl)phosphine (TCEP) enables the use of an S-sulphonate protecting group for thiol-mediated bioconjugation

Abstract: Herein, we demonstrate the effectiveness of the water-friendly Ssulphonate group as an alternative to traditional thiol protecting groups for subsequent deprotection-bioconjugation reactions, under conditions that are compatible with the use of biochemical samples.

Cited by 11 publications

References 25 publications

Glycosyl Bunte Salts: A Class of Intermediates for Sugar Chemistry

Glycosyl Bunte Salts: A Class of Intermediates for Sugar Chemistry

Characterization of Reactions between Water-Soluble Trialkylphosphines and Thiol Alkylating Reagents: Implications for Protein-Conjugation Reactions

Adequate Reducing Conditions Enable Conjugation of Oxidized Peptides to Polymers by One-Pot Thiol Click Chemistry

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