open access publication

Article, 2024

An efficient mRNA display protocol yields potent bicyclic peptide inhibitors for FGFR3c: outperforming linear and monocyclic formats in affinity and stability

Chemical Science, ISSN 2041-6539, 2041-6520, Volume 15, 16, Pages 6122-6129, 10.1039/d3sc04763f

Contributors

Villequey, Camille (Corresponding author) [1] Zurmühl, Silvana S [1] Cramer, Christian N [1] Bhusan, Bhaskar [2] Andersen, Birgitte S 0000-0003-1423-6797 [1] Ren, Qianshen [3] Liu, Haimo [3] Qu, Xinping [3] Yang, Yang [3] Pan, Jia [3] Chen, Qiujia [3] Münzel, Martin [1]

Affiliations

  1. [1] Novo Nordisk (Denmark)
  2. [NORA names: Novo Nordisk; Private Research; Denmark; Europe, EU; Nordic; OECD];
  3. [2] University of Oxford
  4. [NORA names: United Kingdom; Europe, Non-EU; OECD];
  5. [3] Novo Nordisk (China)
  6. [NORA names: China; Asia, East]

Abstract

Macrocyclization has positioned itself as a powerful method for engineering potent peptide drug candidates. Introducing one or multiple cyclizations is a common strategy to improve properties such as affinity, bioavailability and proteolytic stability. Consequently, methodologies to create large libraries of polycyclic peptides by phage or mRNA display have emerged, allowing the rapid identification of binders to virtually any target. Yet, within those libraries, the performance of linear vs. mono- or bicyclic peptides has rarely been studied. Indeed, a key parameter to perform such a comparison is to use a display protocol and cyclization chemistry that enables the formation of all 3 formats in equal quality and diversity. Here, we developed a simple, efficient and fast mRNA display protocol which meets these criteria and can be used to generate highly diverse libraries of thioether cyclized polycyclic peptides. As a proof of concept, we selected peptides against fibroblast growth factor receptor 3c (FGFR3c) and compared the different formats regarding affinity, specificity, and human plasma stability. The peptides with the best KD's and stability were identified among bicyclic peptide hits, further strengthening the body of evidence pointing at the superiority of this class of molecules and providing functional and selective inhibitors of FGFR3c.

Keywords

affinity, bicyclic peptide inhibitors, bicyclic peptides, binder, bioavailability, body, body of evidence points, candidates, chemistry, comparison, concept, criteria, cyclization, cyclization chemistry, display, display protocol, diverse library, diversity, drug candidates, evidence point, fibroblasts, formation, hits, human plasma stability, identification, identification of binders, improved properties, inhibitors, library, mRNA, mRNA display, macrocycle, method, methodology, molecules, multiple cyclizations, parameters, peptide, peptide drug candidates, peptide hits, peptide inhibitor, performance, phage, plasma stability, point, polycyclic peptides, properties, proteolytic stability, protocol, quality, specificity, stability, superiority, target

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