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

Abstract

Following a rational design, a series of macrocyclic ("stapled") peptidomimetics of 10 Panx1, 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 Panx1expressing 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 10 Panx1 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 10 Panx1). 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.

Intercellular Communication in Airway Epithelial Cell Regeneration: Potential Roles of Connexins and Pannexins

“…Gene silencing approaches may also be challenging as airway connexins/pannexins may compensate for each other, as observed, for example, in keratinocytes [125,148]. The ongoing development of connexinand pannexin-targeting peptides may help to address this question [149][150][151]. As the well-differentiated airway epithelium accommodates with low connexin expression, any changes in their expression and/or regulation may alter the repair process.…”

Therapeutic peptides for coronary artery diseases: in silico methods and current perspectives

“…To overcome the problems of the low proteolytic stability of Panx1 inhibitors developed in previous studies, the helical stapling method was applied. As a result, it was reported that the two analogs generated achieved a twofold increase in the inhibitory effect as well as a 30-fold increase in the stability profile under physiological conditions (Lamouroux et al 2023 ).…”