Toolbox of Diverse Linkers for Navigating the Cellular Efficacy Landscape of Stapled Peptides

Yuteng Wu, Amandeep Kaur, Elaine Fowler, Mareike M. Wiedmann, Reginald Young, Warren R.J.D. Galloway, Lasse Olsen, Hannah F. Sore, Anasuya Chattopadhyay, Terence T.L. Kwan, Wenshu Xu, Stephen J. Walsh, Peterson De Andrade, Matej Janecek, Senthil Arumugam, Laura S. Itzhaki, Yu Heng Lau, David R. Spring

Research output: Contribution to journalArticleResearchpeer-review

15 Citations (Scopus)


Stapled peptides have great potential as modulators of protein-protein interactions (PPIs). However, there is a vast landscape of chemical features that can be varied for any given peptide, and identifying a set of features that maximizes cellular uptake and subsequent target engagement remains a key challenge. Herein, we present a systematic analysis of staple functionality on the peptide bioactivity landscape in cellular assays. Through application of a "toolbox" of diversified dialkynyl linkers to the stapling of MDM2-binding peptides via a double-click approach, we conducted a study of cellular uptake and p53 activation as a function of the linker. Minor changes in the linker motif and the specific pairing of linker with peptide sequence can lead to substantial differences in bioactivity, a finding which may have important design implications for peptide-based inhibitors of other PPIs. Given the complexity of the structure-activity relationships involved, the toolbox approach represents a generalizable strategy for optimization when progressing from in vitro binding assays to cellular efficacy studies.

Original languageEnglish
Pages (from-to)526-533
Number of pages8
JournalACS Chemical Biology
Issue number3
Publication statusPublished - 15 Mar 2019
Externally publishedYes

Cite this