PDF(Pubmed)
Abstract：
Coronavirus (SARS-CoV-2) as a global pandemic has attracted the attention of many scientific centers to find the right treatment. We expressed and purified the recombinant receptor-binding domain (RBD) of the SARS-CoV-2 spike (S) protein, and specific RBD aptamers were designed using SELEX method. RNAi targeting nucleocapsid phosphoprotein was synthesized and human lung cells were inoculated with aptamer-functionalized lipid nanoparticles (LNPs) containing RNAi. The results demonstrated that RBD aptamer having KD values of 0.290 nm possessed good affinity. Based on molecular docking and efficacy prediction analysis, siRNA molecule was showed the best action. LNPs were appropriately functionalized by aptamer and contained RNAi molecules. Antiviral assay using q-PCR and ELISA demonstrated that LNP functionalized with 35 µm Apt and containing 30 nm RNAi/ml of cell culture had the best antiviral activity compared to other concentrations. Applied aptamer in the nanocarrier has two important functions. First, it can deliver the drug (RNAi) to the surface of epithelial cells. Second, by binding to the SARS-CoV-2 spike protein, it inhibits the virus entrance into cells. Our data reveal an interaction between the aptamer and the virus, and RNAi targeted the virus RNA. CT scan and the clinical laboratory tests in a clinical case study, a 36-year old man who presented with severe SARS-CoV-2, demonstrated that inhalation of 10 mg Apt-LNPs-RNAi nebulized/day for six days resulted in an improvement in consolidation and ground-glass opacity in lungs on the sixth day of treatment. Our findings suggest the treatment of SARS-CoV-2 infection through inhalation of Aptamer-LNPs-RNAi.
摘要：
冠状病毒（SARS-CoV-2）作为一种全球大流行，吸引了许多科学中心的关注，以找到正确的治疗方法。我们表达并纯化了SARS-CoV-2刺突（S）蛋白的重组受体结合域（RBD），使用SELEX方法设计特异性RBD适体。合成靶向核衣壳磷蛋白的RNAi,并用含有RNAi的适体官能化的脂质纳米颗粒(LNP)接种人肺细胞。结果表明，Kd值为0.290nm的RBD适体具有良好的亲和力。基于分子对接和疗效预测分析,siRNA分子表现出最好的作用。LNP通过适体适当地官能化并且含有RNAi分子。使用q-PCR和ELISA的抗病毒测定表明，与其他浓度相比，用35μmApt官能化并含有30nmRNAi/ml细胞培养物的LNP具有最佳的抗病毒活性。纳米载体中应用的适体具有两个重要功能。首先,它可以将药物（RNAi）传递到上皮细胞表面。第二,通过与SARS-CoV-2尖峰蛋白结合，抑制病毒进入细胞.我们的数据揭示了适体和病毒之间的相互作用，RNAi靶向病毒RNA。临床病例研究中的CT扫描和临床实验室检查，一名36岁的男性出现了严重的SARS-CoV-2，证明吸入10mgApt-LNPs-RNAi/天，持续6天，可在第6天改善肺部的巩固和磨玻璃混浊治疗。我们的发现表明通过吸入适体-LNPs-RNAi治疗SARS-CoV-2感染。
