背景:耐药流感病毒的不断进化凸显了将具有抗流感活性的天然来源和安全的植物化学物质重新用于新型广谱抗流感药物的必要性。
方法:在本研究中,测试了含氮生物碱对A/H1N1和A/H5N1流感病毒的病毒抑制活性。测试生物碱对MDCK的细胞毒性显示出高安全范围(CC50>200µg/ml),允许筛查他们的抗流感潜力。
结果:这里,硫酸阿托品,盐酸毛果芸香碱和秋水仙碱表现出抗H5N1活性,IC50值分别为2.300、0.210和0.111µg/ml,分别。通过测试三种高效生物碱进一步描述了IC50值的验证,基于它们对季节性流感A/H1N1病毒的有效IC50值,显示相当的IC50值为0.204、0.637和0.326微克/毫升,分别。进一步的研究表明,秋水仙碱可以通过主要干扰IAV复制和抑制病毒吸附来抑制病毒感染,而硫酸阿托品和盐酸毛果芸香碱可以直接影响病毒的无细胞杀病毒作用。有趣的是,计算机分子对接研究表明阿托品的能力,毛果芸香碱,和秋水仙碱在A/H1N1和A/H5N1流感病毒的神经氨酸酶的活性位点内正确结合。三种生物碱表现出良好的结合能以及与共结晶配体相似的优异结合模式。另一方面,与体外结果一致,只有秋水仙碱才能正确结合甲型流感病毒(IAV)的M2质子通道。这可能说明秋水仙碱在病毒复制周期的复制阶段的体外抗病毒活性。
结论:本研究强调了包括秋水仙碱在内的生物活性生物碱的抗流感功效。因此,这些生物碱应在体内进一步表征(临床前和临床研究)以开发作为抗IAV药物.
BACKGROUND: The continuous evolution of drug-resistant influenza viruses highlights the necessity for repurposing naturally-derived and safe phytochemicals with anti-influenza activity as novel broad-spectrum anti-influenza medications.
METHODS: In this study, nitrogenous alkaloids were tested for their viral inhibitory activity against influenza A/H1N1 and A/H5N1 viruses. The cytotoxicity of tested alkaloids on MDCK showed a high safety range (CC50 > 200 µg/ml), permitting the screening for their anti-influenza potential.
RESULTS: Herein, atropine sulphate, pilocarpine hydrochloride and colchicine displayed anti-H5N1 activities with IC50 values of 2.300, 0.210 and 0.111 µg/ml, respectively. Validation of the IC50 values was further depicted by testing the three highly effective alkaloids, based on their potent IC50 values against seasonal influenza A/H1N1 virus, showing comparable IC50 values of 0.204, 0.637 and 0.326 µg/ml, respectively. Further investigation suggests that colchicine could suppress viral infection by primarily interfering with IAV replication and inhibiting viral adsorption, while atropine sulphate and pilocarpine hydrochloride could directly affect the virus in a cell-free virucidal effect. Interestingly, the in silico molecular docking studies suggest the abilities of atropine, pilocarpine, and colchicine to bind correctly inside the active sites of the neuraminidases of both influenza A/H1N1 and A/H5N1 viruses. The three alkaloids exhibited good binding energies as well as excellent binding modes that were similar to the co-crystallized ligands. On the other hand, consistent with in vitro results, only colchicine could bind correctly against the M2-proton channel of influenza A viruses (IAVs). This might explicate the in vitro antiviral activity of colchicine at the replication stage of the virus replication cycle.
CONCLUSIONS: This study highlighted the anti-influenza efficacy of biologically active alkaloids including colchicine. Therefore, these alkaloids should be further characterized in vivo (preclinical and clinical studies) to be developed as anti-IAV agents.