In the present study, a novel chimeric peptide was derived from camel lactoferrin designed with a considerable anti-HCV activity and its neutralization mechanism was predicted by molecular modelling tools. A novel anti-HCV peptide derived from camel lactoferrin (cLF36) was designed and expressed it recombinantly in HEK-293-T cells. Anti-viral activity of this peptide was evaluated against hepatitis C virus by Real-time PCR assay in vitro. Finally, to have a better insight into the mode of action of peptide on HCV entry inhibition, we examined the interaction of cLF36 with envelope glycoprotein E2 by molecular dynamic simulation. This chimeric peptide had significant inhibitory effects on both HCV entry (44 µg/mL) and viral replication (88 µg/mL) under in vitro (p > 0.01). Moreover, cLF36 peptide was not toxic to HEK cells as a normal cell at twofold of its anti-viral concentrations for HCV entry and even at concentrations as high as 250 µg/mL exhibited minimal hemolysis (2.5%) against human RBCs (red blood cells). The results of in silico analysis showed that cLF36 interacted with β-sandwich and front layer of E2 protein as two potential CD81 binding sites. We generated and characterized a new camel lactoferrin derived HCV inhibitors. This peptide blocked HCV entry and also intracellular HCV replication in cell culture experiment.

中文翻译：

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骆驼乳铁蛋白衍生的新型嵌合抗HCV肽及其与E2相互作用的分子水平研究

在本研究中，从骆驼乳铁蛋白衍生出一种新颖的嵌合肽，该肽设计具有相当大的抗HCV活性，并且其中和机理可通过分子建模工具预测。设计了一种新的源自骆驼乳铁蛋白的抗HCV肽（cLF36），并在HEK-293-T细胞中重组表达。该肽的抗病毒活性通过体外实时PCR分析评估了对丙型肝炎病毒的抗病毒活性。最后，为了更好地了解肽对HCV进入抑制的作用方式，我们通过分子动力学模拟研究了cLF36与包膜糖蛋白E2的相互作用。这种嵌合肽在体外对HCV进入（44 µg / mL）和病毒复制（88 µg / mL）均具有显着的抑制作用（p> 0.01）。此外，cLF36肽对HEK细胞无害，因为它对于HCV进入来说是其抗病毒浓度的两倍，甚至是正常细胞，甚至在高达250 µg / mL的浓度下，对人RBC（红细胞）的溶血作用也很小（2.5％）。电脑分析的结果表明，cLF36与β三明治和E2蛋白的前层相互作用，是两个潜在的CD81结合位点。我们生成并表征了一种新的骆驼乳铁蛋白衍生的HCV抑制剂。在细胞培养实验中，该肽不仅阻断了HCV的进入，而且阻断了细胞内HCV的复制。电脑分析的结果表明，cLF36与β三明治和E2蛋白的前层相互作用，是两个潜在的CD81结合位点。我们生成并表征了一种新的骆驼乳铁蛋白衍生的HCV抑制剂。在细胞培养实验中，该肽不仅阻断了HCV的进入，而且阻断了细胞内HCV的复制。电脑分析的结果表明，cLF36与β三明治和E2蛋白的前层相互作用，是两个潜在的CD81结合位点。我们生成并表征了一种新的骆驼乳铁蛋白衍生的HCV抑制剂。在细胞培养实验中，该肽不仅阻断了HCV的进入，而且阻断了细胞内HCV的复制。