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

Study of the mechanism of caffeoyl glucopyranoses in inhibiting HIV-1 entry using pseudotyped virus system

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:25μg·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 only 1,2,6-Tri-O-caffeoyl-β-D-glucopyranose （TCGP） and 1,3-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-1 entry inhibitors targeting gp41 and can serve as lead compounds for developing novel anti-HIV-1 microbicides for prevention of sexual HIV-1 transmission.