Zika virus, an arthropod-borne flavivirus, is an emerging healthcare threat worldwide. Zika virus is responsible for severe neurological effects, such as paralytic Guillain-Barrè syndrome, in adults, and also congenital malformations, especially microcephaly. No specific antiviral drugs and vaccines are currently available, and treatments are palliative, but medicinal plants show great potential as natural sources of anti-Zika phytochemicals. This study deals with the investigation of the composition, cytotoxicity, and anti-Zika activity of Punica granatum leaf ethanolic extract, fractions, and phytoconstituents. P. granatum leaves were collected from different areas in Italy and Greece in different seasons. Crude extracts were analyzed and fractionated, and the pure compounds were isolated. The phytochemical and biomolecular fingerprint of the pomegranate leaves was determined. The antiviral activities of the leaf extract, fractions, and compounds were investigated against the MR766 and HPF2013 Zika virus strains in vitro. Both the extract and its fractions were found to be active against Zika virus infection. Of the compounds isolated, ellagic acid showed particular anti-Zika activities, with EC50 values of 30.86 µM for MR766 and 46.23 µM for HPF2013. The mechanism of action was investigated using specific antiviral assays, and it was demonstrated that ellagic acid was primarily active as it prevented Zika virus infection and was able to significantly reduce Zika virus progeny production. Our data demonstrate the anti-Zika activity of pomegranate leaf extract and ellagic acid for the first time. These findings identify ellagic acid as a possible anti-Zika candidate compound that can be used for preventive and therapeutic interventions.

中文翻译：

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石榴叶乙醇提取物和鞣花酸作为寨卡病毒感染的抑制剂

寨卡病毒是一种节肢动物传播的黄病毒，是全球新兴的医疗保健威胁。寨卡病毒会导致严重的神经系统影响，例如成人麻痹性格林-巴雷综合征，以及先天性畸形，尤其是小头畸形。目前没有特定的抗病毒药物和疫苗可用，治疗方法是姑息性的，但药用植物作为抗寨卡病毒植物化学物质的天然来源显示出巨大的潜力。本研究涉及石榴叶乙醇提取物、馏分和植物成分的组成、细胞毒性和抗寨卡病毒活性的调查。P. granatum 叶子是在不同季节从意大利和希腊的不同地区收集的。对粗提取物进行分析和分级，分离出纯化合物。测定了石榴叶的植物化学和生物分子指纹。在体外研究了叶提取物、级分和化合物对 MR766 和 HPF2013 寨卡病毒株的抗病毒活性。发现提取物及其组分均具有抗寨卡病毒感染的活性。在分离的化合物中，鞣花酸显示出特殊的抗寨卡病毒活性，MR766 的 EC50 值为 30.86 µM，HPF2013 的 EC50 值为 46.23 µM。使用特定的抗病毒试验研究了作用机制，证明鞣花酸主要具有活性，因为它可以预防寨卡病毒感染并能够显着减少寨卡病毒后代的产生。我们的数据首次证明了石榴叶提取物和鞣花酸的抗寨卡病毒活性。