日本蛇菰中咖啡酰基葡萄糖类化合物抑制HIV-1进入的机制

目的：研究从日本蛇菰中分离的咖啡酰基葡萄糖类化合物抑制HIV-1进入的作用机制。方法：利用HIV-1假病毒体系，并结合针对gｐ41包膜蛋白的ELISA以及分子对接方法，对日本蛇菰分离的咖啡酰基葡萄糖类化合物抑制HIV-1 进入机制进行研究。结果：我们首先用HIV-1 Env假病毒来检测六个咖啡酰基葡萄糖类化合物（终浓度：25 µg・ml^-1）的抑制活性，发现1,2,6-三-Ｏ-咖啡酰基-β-D-吡喃葡萄糖（TCGP）和1,3-双-Ｏ-咖啡酰基-4-Ｏ-没食子酰-β-D-吡喃葡萄糖（DCGGP）具有较好的抑制病毒进入靶细胞的作用。进一步量效关系研究表明，两个化合物的病毒抑制活性呈剂量依赖性，它们的IＣ₅₀值分别是（5.5±0.2）和（5.3±0.1）µg・ml^-1。这两个化合物的抑制活性与其抑制gp41六螺旋束结构形成的机制有关。分子对接表明，它们均能靶向gp41上N-螺旋三聚体的靶穴位置。结论：1,2,6-三-Ｏ-咖啡酰基-β-D-吡喃葡萄糖（TCGP）和1,3-双-Ｏ-咖啡酰基-4-Ｏ-没食子酰-β-D-吡喃葡萄糖（DCGGP）可以较好的抑制HIV-1 Env假病毒感染靶细胞，可作为先导化合物来研发抗HIV-1新药。

Objective: To investigate the inhibitory activities of caffeoyl glucopyranoses purified from Balanophora japonica Makino on HIV entry and their mechanism. Methods: HIV-1 Env pseudovirus was used to evaluate the anti-HIV-1 activity of those compounds. ELISA and molecular docking were used to study the mechanism of the actions of the active compounds. Results: We used the HIV-1 Env pseudovirus to test the anti-HIV-1 activity of the six phenolic compounds (final concentration 25 µg/ml), and found that only1,2,6-Tri-O-caffeoyl-β-D-glucopyranose (TCGP) andl,3-Di-O-caffeoyl-4-O-galloyl-β-D-glucopyranose (DCGGP) could effectively inhibit the entry of HIV-1 Env pseudovirus into the target cells in a dose-dependent manner, with IC₅₀ values of 5.5±0.2 and 5.3±0.1 µg/ml, respectively. These two compounds could also blocked the gp41 six-helix bundle formation. Molecular docking analysis suggested that they might bind to the hydrophobic cavity of the gp41 N-trimeric coiled-coil. Conclusion: TCGP and DCGGP are potent HIV-1entry inhibitors targeting gp41 and can serve as lead compounds for developing novel anti-HIV-1 microbicides for prevention of sexual HIV-1 transmission.