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Angus
P. R. Johnston (1629103)

Athina Anastasaki (1305825)

David M. Haddleton (1273797)

John F. Quinn (1372680)

Kristian Kempe (1308939)

Matthew R. Owen (1629106)

Michael R. Whittaker (1356420)

Paul Wilson (146891)

Philip J. Hogg (1629097)

Sarah K. Mann (1629100)

Thomas P. Davis (521458)

Paul Wilson

Athina Anastasaki

Matthew R. Owen

Kristian Kempe

David M. Haddleton

Sarah K. Mann

Angus P. R. Johnston

John F. Quinn

Michael R. Whittaker

Philip J. Hogg

Thomas P. Davis

Crossref

Journal of the American Chemical Society

Organic Arsenicals As Efficient and Highly Specific Linkers for Protein/Peptide–Polymer Conjugation

The entropy-driven affinity of trivalent (in)organic arsenicals for closely spaced dithiols has been exploited to develop a novel route to peptide/protein-polymer conjugation. A trivalent arsenous acid (As(III)) derivative (1) obtained from p-arsanilic acid (As(V)) was shown to readily undergo conjugation to the therapeutic peptide salmon calcitonin (sCT) via bridging of the Cys1-Cys7 disulfide, which was verified by RP-HPLC and MALDI-ToF-MS. Conjugation was shown to proceed rapidly (t < 2 min) in situ and stoichiometrically through sequential reduction-conjugation protocols, therefore exhibiting conjugation efficiencies equivalent to those reported for the current leading disulfide-bond targeting strategies. Furthermore, using bovine serum albumin as a model protein, the trivalent organic arsenical 1 was found to demonstrate enhanced specificity for disulfide-bond bridging in the presence of free cysteine residues relative to established maleimide functional reagents. This specificity represents a shift toward potential orthogonality, by clearly distinguishing between the reactivity of mono- and disulfide-derived (vicinal or neighbors-through-space) dithiols. Finally, p-arsanilic acid was transformed into an initiator for aqueous single electron-transfer living radical polymerization, allowing the synthesis of hydrophilic arsenic-functional polymers which were shown to exhibit negligible cytotoxicity relative to a small molecule organic arsenical, and an unfunctionalized polymer control. Poly(poly[ethylene glycol] methyl ether acrylate) (PPEGA(480), DPn = 10, M-n,M-NMR = 4900 g.mol(-1), D = 1.07) possessing a pentavalent arsenic acid (As(V)) alpha-chain end was transformed into trivalent As(III) post-polymerization via initial reduction by biological reducing agent glutathione (GSH), followed by binding of GSH. Conjugation of the resulting As(III)-functional polymer to sCT was realized within 35 min as indicated by RP-HPLC and verified later by thermodynamically driven release of sCT, from the conjugate, in the presence of strong chelating reagent ethanedithiol

Wilson, Paul

Anastasaki, Athina

Owen, Matthew R.

Kempe, Kristian

Haddleton, David M.

Mann, Sarah K.

Johnston, Angus P. R.

Quinn, John F.

Whittaker, Michael R.

Hogg, Philip J.

Davis, Thomas P.

University of Queensland eSpace

Organic arsenicals as efficient and highly specific linkers for protein/peptide-polymer conjugation

The entropy-driven affinity of trivalent (in)organic arsenicals for closely spaced dithiols has been exploited to develop a novel route to peptide/protein–polymer conjugation. A trivalent arsenous acid (As(III)) derivative (1) obtained from p-arsanilic acid (As(V)) was shown to readily undergo conjugation to the therapeutic peptide salmon calcitonin (sCT) via bridging of the Cys1-Cys7 disulfide, which was verified by RP-HPLC and MALDI-ToF-MS. Conjugation was shown to proceed rapidly (t < 2 min) in situ and stoichiometrically through sequential reduction–conjugation protocols, therefore exhibiting conjugation efficiencies equivalent to those reported for the current leading disulfide-bond targeting strategies. Furthermore, using bovine serum albumin as a model protein, the trivalent organic arsenical 1 was found to demonstrate enhanced specificity for disulfide-bond bridging in the presence of free cysteine residues relative to established maleimide functional reagents. This specificity represents a shift toward potential orthogonality, by clearly distinguishing between the reactivity of mono- and disulfide-derived (vicinal or neighbors-through-space) dithiols. Finally, p-arsanilic acid was transformed into an initiator for aqueous single electron-transfer living radical polymerization, allowing the synthesis of hydrophilic arsenic-functional polymers which were shown to exhibit negligible cytotoxicity relative to a small molecule organic arsenical, and an unfunctionalized polymer control. Poly(poly[ethylene glycol] methyl ether acrylate) (PPEGA480, DPn = 10, Mn,NMR = 4900 g·mol–1, Đ = 1.07) possessing a pentavalent arsenic acid (As(V)) α-chain end was transformed into trivalent As(III) post-polymerization via initial reduction by biological reducing agent glutathione (GSH), followed by binding of GSH. Conjugation of the resulting As(III)-functional polymer to sCT was realized within 35 min as indicated by RP-HPLC and verified later by thermodynamically driven release of sCT, from the conjugate, in the presence of strong chelating reagent ethanedithiol

Warwick Research Archives Portal Repository

Organic arsenicals As efficient and highly specific linkers for protein/peptide–polymer conjugation

The entropy-driven affinity of trivalent
(in)­organic arsenicals
for closely spaced dithiols has been exploited to develop a novel
route to peptide/protein–polymer conjugation. A trivalent arsenous
acid (As­(III)) derivative (1) obtained from p-arsanilic acid (As­(V)) was shown to readily undergo conjugation
to the therapeutic peptide salmon calcitonin (sCT) via bridging of
the Cys1-Cys7 disulfide, which was verified
by RP-HPLC and MALDI-ToF-MS. Conjugation was shown to proceed rapidly
(t < 2 min) in situ and stoichiometrically
through sequential reduction–conjugation protocols, therefore
exhibiting conjugation efficiencies equivalent to those reported for
the current leading disulfide-bond targeting strategies. Furthermore,
using bovine serum albumin as a model protein, the trivalent organic
arsenical 1 was found to demonstrate enhanced specificity
for disulfide-bond bridging in the presence of free cysteine residues
relative to established maleimide functional reagents. This specificity
represents a shift toward potential orthogonality, by clearly distinguishing
between the reactivity of mono- and disulfide-derived (vicinal or
neighbors-through-space) dithiols. Finally, p-arsanilic
acid was transformed into an initiator for aqueous single electron-transfer
living radical polymerization, allowing the synthesis of hydrophilic
arsenic-functional polymers which were shown to exhibit negligible
cytotoxicity relative to a small molecule organic arsenical, and an
unfunctionalized polymer control. Poly­(poly­[ethylene glycol] methyl
ether acrylate) (PPEGA480, DPn = 10, Mn,NMR = 4900 g·mol–1, Đ = 1.07) possessing a
pentavalent arsenic acid (As­(V)) α-chain end was transformed
into trivalent As­(III) post-polymerization via initial reduction by
biological reducing agent glutathione (GSH), followed by binding of
GSH. Conjugation of the resulting As­(III)-functional polymer to sCT
was realized within 35 min as indicated by RP-HPLC and verified
later by thermodynamically driven release of sCT, from the conjugate,
in the presence of strong chelating reagent ethanedithiol

FigShare

Organic
Arsenicals As Efficient and Highly Specific
Linkers for Protein/Peptide–Polymer Conjugation

https://figshare.com/articles/Organic_Arsenicals_As_Efficient_and_Highly_Specific_Linkers_for_Protein_Peptide_Polymer_Conjugation/2182051