Structure-Based Design and Synthesis of Stapled 10Panx1 Analogues for Use in Cardiovascular Inflammatory Diseases

• DOI: 10.1021/acs.jmedchem.3c01116
• Publication Date: 2023-09-13T15:42:31Z
• Authors: Arthur Lamouroux; Malaury Tournier; Debora Iaculli; Anne Caufriez; Olga M. Rusiecka; Charlotte Martin; Viviane Bes; Laureano E. Carpio; Yana Girardin; Remy Loris; Andrés Tabernilla; Filippo Molica; Rafael Gozalbes; María D. Mayán; Mathieu Vinken; Brenda R. Kwak; Steven Ballet
• Keywords: Adenosine Triphosphate; Cardiovascular Diseases; info:eu-repo/classification/ddc/616.07; Cell Line, Tumor; Animals; Humans; Endothelial Cells; Peptides; Connexins

Following a rational design, a series of macrocyclic ("stapled") peptidomimetics of 10Panx1, the most established peptide inhibitor of Pannexin1 (Panx1) channels, were developed and synthesized. Two macrocyclic analogues SBL-PX1-42 and SBL-PX1-44 outperformed the linear native peptide. During in vitro adenosine triphosphate (ATP) release and Yo-Pro-1 uptake assays in a Panx1-expressing tumor cell line, both compounds were revealed to be promising bidirectional inhibitors of Panx1 channel function, able to induce a two-fold inhibition, as compared to the native 10Panx1 sequence. The introduction of triazole-based cross-links within the peptide backbones increased helical content and enhanced in vitro proteolytic stability in human plasma (>30-fold longer half-lives, compared to 10Panx1). In adhesion assays, a "double-stapled" peptide, SBL-PX1-206 inhibited ATP release from endothelial cells, thereby efficiently reducing THP-1 monocyte adhesion to a TNF-α-activated endothelial monolayer and making it a promising candidate for future in vivo investigations in animal models of cardiovascular inflammatory disease.