Rosmarinic acid inhibits Rift Valley fevervirus: in vitro, computational and analyticalstudies: supplementary materials

Aim: The antiviral potentials of rosmarinic acid (RA) against Rift Valley fever (RVF) virus were investigated.Methods: Antiviral activity was investigated by evaluating the reduction in the viral infectivity titer.Computational and LC–MS studies were performed for investigating the mechanism of action. This isvia testing the interaction between RA and its major metabolite with the key infectivity proteins anddetermination of RA cellular permeability. Results: A superior reduction in RVF infectivity titer (45.5%)was observed when RA was applied post-infection compared to 17.7% reduction following its applicationbefore infection in addition to time-dependent inactivation kinetics. Recorded data showed in-silicoinhibitory potential of RA and its metabolite against RVFV cap-binding protein and glycoprotein C, whichare integral for viral transcription. LC–MS revealed cellular inclusion of RA, supporting its intracellularviral interaction. Conclusion: These antiviral potentials might suggest a promising foundation for futureanti-RVF drug development.研究目的：探究迷迭香酸（rosmarinic acid, RA）对抗裂谷热（Rift Valley fever, RVF）病毒的抗病毒潜能。 实验方法：通过评估病毒感染性滴度的降幅探究其抗病毒活性；同时开展计算机模拟及液相色谱-质谱联用（LC–MS）实验以解析其作用机制，具体包括检测迷迭香酸与其主要代谢产物与病毒关键感染性蛋白的相互作用，以及测定迷迭香酸的细胞通透性。 实验结果：相较于感染前给药仅实现17.7%的病毒滴度降幅，感染后给药时迷迭香酸可使裂谷热病毒感染性滴度降低45.5%，降幅更为显著，同时呈现出时间依赖性的灭活动力学特征。记录的数据显示，在计算机模拟（in silico）层面，迷迭香酸及其代谢产物对裂谷热病毒帽结合蛋白（cap-binding protein）与糖蛋白C（glycoprotein C）具有潜在抑制活性，而这两类蛋白是病毒转录过程不可或缺的组分。液相色谱-质谱联用实验证实了迷迭香酸可被细胞摄取，为其在细胞内与病毒发生相互作用提供了佐证。 研究结论：本研究揭示的抗病毒活性可为未来抗裂谷热病毒药物的研发提供极具前景的理论基础。