Oral Presentation The 16th Australian Peptide Conference 2025

Discovery of Selective Cyclic D-Sulfopeptide Ligands of the Chemokine CCL22 via Mirror-Image mRNA Display with Genetic Reprogramming (128344)

Belinda B. Zhang 1 2 , Katriona Harrison 1 2 , Yichen Zhong 1 2 , Joshua W. C. Maxwell 1 2 , Daniel J. Ford 1 2 , Liam P. Calvey 1 2 , Sean S. So 3 , Francis C. Peterson 4 5 , Brian F. Volkman 4 5 , Martin J. Stone 3 , Ram Prasad Bhusal 3 , Sameer S. Kulkarni 1 2 , Richard J. Payne 1 2
  1. School of Chemistry, The University of Sydney, Sydney, NSW, Australia
  2. Australian Research Council Centre of Excellence for Innovations in Peptide and Protein Science, The University of Sydney, NSW
  3. Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia
  4. Department of Biochemistry, Medical College of Wisconsin, Milwaukee, Wisconsin, USA
  5. Protein Foundry, Milwaukee, Wisconsin, USA

Inflammation is driven by chemokines – small proteins involved in recruiting leukocytes through interactions with specific cell-surface receptors.1 CCL22 is a chemokine that plays a crucial role in inflammatory diseases such as asthma and atopic dermatitis, thus inhibition of CCL22 presents an emerging therapeutic strategy.2 In this work, we discovered cyclic D‑sulfopeptide inhibitors of CCL22 using mirror-image mRNA display with genetic reprogramming. We first synthesised mirror-image D-CCL22, which was subsequently screened against a cyclic peptide library composed of all L‑amino acids, including reprogrammed L‑sulfotyrosine to mimic this post-translational modification present on native chemokine receptors. The most enriched peptides were prepared as mirror-image D-ligands and assessed for binding against native L‑CCL22. We found that the most potent ligand, a plasma-stable D-cyclic peptide with four D-sulfotyrosine residues, showed nanomolar affinity for CCL22, nanomolar inhibition of CCL22 signaling via CCR4, and remarkable selectivity over other chemokines. Thus, this work highlights the potential of mirror-image mRNA display as a technology for discovering proteolytically stable D-peptide inhibitors of protein-protein interactions relevant across a range of therapeutic indications.3

  1. Hughes, C. E.; Nibbs, R. J. B. A Guide to Chemokines and Their Receptors. FEBS J. 2018, 285 (16), 2944-2971.
  2. Kufareva, I.; Salanga, C. L.; Handel, T. M. Chemokine and Chemokine Receptor Structure and Interactions: Implications for Therapeutic Strategies. Immunol. Cell Biol. 2015, 93 (4), 372-383.
  3. Zhang, B. B.; Harrison, K.; Zhong, Y.; Maxwell, J. W. C.; Ford, D. J.; Calvey, L. P.; So, S. S.; Peterson, F. C.; Volkman, B. F.; Stone, M. J.; Bhusal, R. P.; Kulkarni, S. S.; Payne, R. J. Discovery of Selective Cyclic D-Sulfopeptide Ligands of the Chemokine CCL22 via Mirror-Image mRNA Display with Genetic Reprogramming. J. Am. Chem. Soc. 2024, 146 (50), 34253-34259.