State Key Laboratory of Bioactive Substance and Function of Natual Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.
Beijing Key Laboratory of Active Substances Discovery and Druggability Evaluation, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.
FEBS J. 2023 Nov;290(22):5353-5372. doi: 10.1111/febs.16920. Epub 2023 Aug 17.
We previously identified glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as one of the cyclic adenosine diphosphoribose (cADPR)'s binding proteins and found that GAPDH participates in cADPR-mediated Ca release from endoplasmic reticulum via ryanodine receptors (RyRs). Here, we aimed to chemically synthesise and pharmacologically characterise novel cADPR analogues. Based on the simulated cADPR-GAPDH complex structure, we performed the structure-based drug screening, identified several small chemicals with high docking scores to cADPR's binding pocket in GAPDH and showed that two of these compounds, C244 and C346, are potential cADPR antagonists. We further synthesised several analogues of C346 and found that its analogue, G42, also mobilised Ca release from lysosomes. G42 alkalised lysosomal pH and inhibited autophagosome-lysosome fusion. Moreover, G42 markedly inhibited Zika virus (ZIKV, a flavivirus) or murine hepatitis virus (MHV, a β-coronavirus) infections of host cells. These results suggest that G42 inhibits virus infection, likely by triggering lysosomal Ca mobilisation and inhibiting autophagy.
我们之前鉴定出甘油醛 3-磷酸脱氢酶（GAPDH）是环腺苷二磷酸核糖（cADPR）的结合蛋白之一，并发现 GAPDH 通过肌质网 Ryanodine 受体（RyRs）参与 cADPR 介导的 Ca 释放。在此，我们旨在通过化学合成和药理学方法来鉴定新型 cADPR 类似物。基于模拟的 cADPR-GAPDH 复合物结构，我们进行了基于结构的药物筛选，鉴定出几种与 GAPDH 中 cADPR 结合口袋具有高对接评分的小分子化合物，并表明其中两种化合物 C244 和 C346 是潜在的 cADPR 拮抗剂。我们进一步合成了 C346 的几种类似物，发现其类似物 G42 也能从溶酶体中动员 Ca 释放。G42 使溶酶体 pH 碱化并抑制自噬体-溶酶体融合。此外，G42 明显抑制寨卡病毒（ZIKV，黄病毒）或鼠肝炎病毒（MHV，β-冠状病毒）感染宿主细胞。这些结果表明，G42 通过触发溶酶体 Ca 动员和抑制自噬来抑制病毒感染。
