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Twin disulfides for orthogonal disulfide pairing and the directed folding of multicyclic peptides


Multicyclic peptides are emerging as an exciting platform for drug and targeted ligand discovery owing to their expected greater target affinity/selectivity/stability versus linear or monocyclic peptides. However, although the precise pairing of cysteine residues in proteins is routinely achieved in nature, the rational pairing of cysteine residues within polypeptides is a long-standing challenge for the preparation of multicyclic species containing several disulfide bridges. Here, we present an efficient and straightforward approach for directing the intermolecular and intramolecular pairing of cysteine residues within peptides using a minimal CXC motif. Orthogonal disulfide pairing can be exploited in complex redox media to rationally produce dimeric peptides and bi/tricyclic peptides from fully reduced peptides containing 1–6 cysteine residues. This strategy, which does not rely on extensive manipulation of the primary sequence, post-translational modification or protecting groups, should greatly benefit the development of multicyclic peptide therapeutics and targeting ligands.

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Figure 1: Equilibrium of peptides containing a CXC motif in a redox buffer (RSH/RSSR).
Figure 2: Selective formation and cleavage of intermolecular disulfide bonds.
Figure 3: Directed folding of peptides containing four cysteine residues to bicyclic structures.
Figure 4: Directed folding of peptides containing six cysteine residues to tricyclic structures.


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C.W. acknowledges a postdoctoral fellowship from the ETHZ (FEL-09 10-1).

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C.W. designed, performed and analysed experiments. J-C.L. and M.A.G. designed and analysed experiments. All authors contributed to writing the manuscript.

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Correspondence to Jean-Christophe Leroux or Marc A. Gauthier.

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The authors declare no competing financial interests.

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Wu, C., Leroux, JC. & Gauthier, M. Twin disulfides for orthogonal disulfide pairing and the directed folding of multicyclic peptides. Nature Chem 4, 1044–1049 (2012).

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