Necroptosis is reported to play a critical role in contributing to a variety of human pathologies. The benzoxazepinone GSK′772 is a potent necroptosis inhibitor optimized using a hit from a DNA-encoded library, which is currently in phase II clinical trials for psoriasis, rheumatoid arthritis, and ulcerative colitis. In the present study, the bioisosterism strategy was applied to replace the amide and benzene ring of GSK′772 based on the co-crystal structure of GSK′772 with its binding target RIPK1. As a result, the novel thio-benzoxazepinones exhibited higher anti-necroptosis activity in a human HT-29 cell necroptosis model. The effect on anti-necroptosis activity by the chirality was significantly reduced in the thio-benzoxazepinones, which was explained by the ligand conformation calculation. Among these analogues, compound 11 (S) and 12 (R) specifically inhibited necroptosis rather than apoptosis with EC₅₀ values of 2.8 and 22.6 nM. They blocked necrosome formation by inhibiting the phosphorylation of RIPK1, RIPK3 and MLKL in necroptotic cells. Collectively, the highly potent thio-benzoxazepinones represent promising lead structures for further development of necroptosis-related diseases.

据报道，坏死性凋亡在导致各种人类病理方面发挥着关键作用。苯并恶杂酮 GSK'772 是一种有效的坏死性凋亡抑制剂，使用来自 DNA 编码文库的命中进行了优化，该文库目前正处于银屑病、类风湿性关节炎和溃疡性结肠炎的 II 期临床试验中。在本研究中，基于 GSK'772 与其结合靶标 RIPK1 的共晶结构，应用生物等排策略取代 GSK'772 的酰胺和苯环。因此，新型硫代苯并氧杂氮杂酮在人 HT-29 细胞坏死性凋亡模型中表现出更高的抗坏死性凋亡活性。手性对抗坏死性凋亡活性的影响在 thio-benzoxazep​​inones 中显着降低，这可以通过配体构象计算来解释。在这些类似物中，化合物11 (S) 和12 (R)以2.8 和22.6 nM 的EC ₅₀值特异性抑制坏死性凋亡而不是细胞凋亡。他们通过抑制坏死细胞中 RIPK1、RIPK3 和 MLKL 的磷酸化来阻断坏死体的形成。总的来说，高效的硫代苯并氧杂氮杂酮代表了用于进一步发展坏死性凋亡相关疾病的有希望的先导结构。