根据理性设计的方法，我们开发并合成了一系列大环（“stapled”）类肽模拟物，以作为Pannexin1（Panx1）通道的最佳已知肽抑制剂10Panx1的替代品。其中两个大环类似物SBL-PX1-42和SBL-PX1-44的性能优于线性天然肽。通过在Panx1表达的肿瘤细胞系中进行体外腺苷三磷酸（ATP）释放和Yo-Pro-1摄取实验，两种化合物被证明是有希望的Panx1通道双向抑制剂，能够诱导两倍于天然10Panx1序列的抑制效果。在肽背骨中引入了三唑交联，增加了肽的螺旋结构含量，并提高了体外人血浆中的蛋白酶稳定性（半衰期增加了30倍，相比于10Panx1）。在粘附实验中，一种“双stapled”肽SBL-PX1-206能够抑制内皮细胞中的ATP释放，从而有效减少THP-1单核细胞对TNF-α激活的内皮单层的粘附，为未来在心血管炎症疾病的动物模型中进行体内研究提供了有希望的候选物。

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.