Sakai Alexandra, Singh Gagandeep, Khoshbakht Mahsa, Bittner Scott, Löhr Christiane V, Diaz-Tapia Randy, Warang Prajakta, White Kris, Luo Luke Le, Tolbert Blanton, Blanco Mario, Chow Amy, Guttman Mitchell, Li Cuiping, Bao Yiming, Ho Joses, Maurer-Stroh Sebastian, Chatterjee Arnab, Chanda Sumit, García-Sastre Adolfo, Schotsaert Michael, Teijaro John R, Moulton Hong M, Stein David A
Scripps Research Institute, La Jolla, CA 92037, USA.
Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
Mol Ther Nucleic Acids. 2024 Sep 10;35(4):102331. doi: 10.1016/j.omtn.2024.102331. eCollection 2024 Dec 10.
Further development of direct-acting antiviral agents against human SARS-CoV-2 infections remains a public health priority. Here, we report that an antisense peptide-conjugated morpholino oligomer (PPMO) named 5'END-2, targeting a highly conserved sequence in the 5' UTR of SARS-CoV-2 genomic RNA, potently suppressed SARS-CoV-2 growth and . In HeLa-ACE 2 cells, 5'END-2 produced IC values of between 40 nM and 1.15 μM in challenges using six genetically disparate strains of SARS-CoV-2, including JN.1. , using K18-hACE2 mice and the WA-1/2020 virus isolate, two doses of 5'END-2 at 10 mg/kg, administered intranasally on the day before and the day after infection, produced approximately 1.4 log10 virus titer reduction in lung tissue at 3 days post-infection. Under a similar dosing schedule, intratracheal administration of 1.0-2.0 mg/kg 5'END-2 produced over 3.5 log10 virus growth suppression in mouse lungs. Electrophoretic mobility shift assays characterized specific binding of 5'END-2 to its complementary target RNA. Furthermore, using reporter constructs containing SARS-CoV-2 5' UTR leader sequence, in an in-cell system, we observed that 5'END-2 could interfere with translation in a sequence-specific manner. The results demonstrate that direct pulmonary delivery of 5'END-2 PPMO is a promising antiviral strategy against SARS-CoV-2 infections and warrants further development.
进一步研发针对人类严重急性呼吸综合征冠状病毒2（SARS-CoV-2）感染的直接作用抗病毒药物仍然是公共卫生领域的一项优先任务。在此，我们报告一种名为5'END-2的反义肽缀合吗啉代寡聚物（PPMO），它靶向SARS-CoV-2基因组RNA 5'非翻译区（UTR）中的一个高度保守序列，能有效抑制SARS-CoV-2的生长。在HeLa-ACE 2细胞中，在使用六种基因不同的SARS-CoV-2毒株（包括JN.1）进行的挑战实验中，5'END-2产生的半数抑制浓度（IC）值在40 nM至1.15 μM之间。使用K18-hACE2小鼠和WA-1/2020病毒分离株，在感染前一天和感染后一天经鼻给予两剂10 mg/kg的5'END-2，在感染后3天，肺组织中的病毒滴度降低了约1.4个对数10。在类似的给药方案下，气管内给予1.0 - 2.0 mg/kg的5'END-2可使小鼠肺部的病毒生长抑制超过3.5个对数10。电泳迁移率变动分析表明5'END-2与其互补靶RNA有特异性结合。此外，在细胞内系统中，使用含有SARS-CoV-2 5'UTR前导序列的报告基因构建体，我们观察到5'END-2可以序列特异性方式干扰翻译。结果表明，直接肺部递送5'END-2 PPMO是一种有前景的抗SARS-CoV-2感染的抗病毒策略，值得进一步研发。
