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Synthesis of all-hydrocarbon stapled α-helical peptides by ring-closing olefin metathesis

Nature Protocols volume 6, pages 761–771 (2011)Cite this article

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Abstract

This protocol provides a detailed procedure for the preparation of stapled α-helical peptides, which have proven their potential as useful molecular probes and as next-generation therapeutics. Two crucial features of this protocol are (i) the construction of peptide substrates containing hindered α-methyl, α-alkenyl amino acids and (ii) the ring-closing olefin metathesis (RCM) of the resulting resin-bound peptide substrates. The stapling systems described in this protocol, namely bridging one or two turns of an α-helix, are highly adaptable to most peptide sequences, resulting in favorable RCM kinetics, helix stabilization and promotion of cellular uptake.

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Figure 1: Schematic representation of three stapled peptides.
Figure 2: Design of stapled peptides.
Figure 3: Overall synthetic scheme for the preparation of stapled peptides using Fmoc-based solid-phase peptide synthesis.
Figure 4
Figure 5: HPLC profiles of RCM reactions of a resin-bound model peptide (Fmoc-EWAS5TAAS5KFLAAHA8,18) monitored at 280 nm after (a) 0 min, (b) 10 min, (c) 30 min, (d) 60 min, (e) 120 min and (f) 240 min.
Figure 6

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Acknowledgements

This research was supported by the Harvard and Dana-Farber Program in Cancer Chemical Biology. T.N.G. is grateful for a fellowship from Deutsche Akademie der Naturforscher Leopoldina (LPDS 2009-2).

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Author notes

  1. Young-Woo Kim

    Present address: Present address: College of Pharmacy, Dongguk University, Goyang-si, Seoul, Korea (Y.-W.K.).,

Authors and Affiliations

  1. Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA

    Young-Woo Kim, Tom N Grossmann & Gregory L Verdine

  2. Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA

    Young-Woo Kim, Tom N Grossmann & Gregory L Verdine

  3. Program in Cancer Chemical Biology, Dana-Farber Cancer Institute, Boston, Massachusetts, USA

    Young-Woo Kim & Gregory L Verdine

  4. Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts, USA

    Gregory L Verdine

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G.L.V. directed the research and, with Y.-W.K., conceptualized the experiments. Y.-W.K. performed the experiments. All authors discussed the results and wrote the manuscript.

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Correspondence to Gregory L Verdine.

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Competing interests

G.L.V. is a shareholder in and a paid consultant of Aileron Therapeutics, which has been granted a license from Harvard University and the Dana-Farber Cancer Institute to develop the stapled peptide technology.

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Kim, YW., Grossmann, T. & Verdine, G. Synthesis of all-hydrocarbon stapled α-helical peptides by ring-closing olefin metathesis. Nat Protoc 6, 761–771 (2011). https://doi.org/10.1038/nprot.2011.324

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