猪流行性腹泻病毒(PEDV),属于α-冠状病毒,是猪流行性腹泻(PED)的病原体。目前,从现有的PEDV疫苗的保护是无效的。因此,抗PEDV化合物应进行研究。小檗碱(BBM),Fangchinoline(FAN),和(+)-Fangchinoline(+FAN),是从天然药用植物中提取的双苄基异喹啉生物碱的类型。这些双苄基异喹啉生物碱具有多种生物活性,包括抗病毒,抗癌,和抗炎特性。在这项研究中,我们发现BBM,风扇,和+FAN抑制PEDV活性,50%抑制浓度为9.00µM,3.54µM,和4.68µM,分别。此外,这些生物碱可以在体外降低PEDV-N蛋白水平和病毒滴度。添加时间测定结果表明,这些生物碱主要抑制PEDV进入。我们还发现BBM的抑制作用,风扇,PEDV和+FAN依赖于通过抑制溶酶体酸化来降低组织蛋白酶L(CTSL)和组织蛋白酶B(CTSB)的活性。一起来看,这些结果表明,BBM,风扇,和+FAN是有效的抗PEDV天然产物,可防止PEDV进入,可能被认为是新型抗病毒药物。
The porcine epidemic diarrhea virus (PEDV), belonging to the α-coronavirus, is the causative agent of porcine epidemic diarrhea (PED). Presently, protection from the existing PEDV vaccine is not effective. Therefore, anti-PEDV compounds should be studied. Berbamine (BBM), Fangchinoline (FAN), and (+)-Fangchinoline (+FAN), are types of bis-benzylisoquinoline alkaloids that are extracted from natural medicinal plants. These bis-benzylisoquinoline alkaloids have various biological activities, including antiviral, anticancer, and anti-inflammatory properties. In this study, we found that BBM, FAN, and +FAN suppressed PEDV activity with a 50% inhibitory concentration of 9.00 µM, 3.54 µM, and 4.68 µM, respectively. Furthermore, these alkaloids can decrease the PEDV-N protein levels and virus titers in vitro. The time-of-addition assay results showed that these alkaloids mainly inhibit PEDV entry. We also found that the inhibitory effects of BBM, FAN, and +FAN on PEDV rely on decreasing the activity of Cathepsin L (CTSL) and Cathepsin B (CTSB) by suppressing lysosome acidification. Taken together, these results indicated that BBM, FAN, and +FAN were effective anti-PEDV natural products that prevented PEDV entry and may be considered novel antiviral drugs.