Anti-SARS-CoV-2 gapmer antisense oligonucleotides targeting the main protease region of viral RNA

IF 4.5 2区 医学 Q1 PHARMACOLOGY & PHARMACY Antiviral research Pub Date : 2024-08-23 DOI:10.1016/j.antiviral.2024.105992
Masako Yamasaki , Wakana Saso , Takuya Yamamoto , Masayoshi Sato , Hiroko Takagi , Tetsuya Hasegawa , Yuji Kozakura , Hiroyuki Yokoi , Hirofumi Ohashi , Kana Tsuchimoto , Rina Hashimoto , Shuetsu Fukushi , Akihiko Uda , Masamichi Muramatsu , Kazuo Takayama , Ken Maeda , Yoshimasa Takahashi , Tsuyoshi Nagase , Koichi Watashi
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Abstract

Given the worldwide risk for the outbreak of emerging/re-emerging respiratory viruses, establishment of new antiviral strategies is greatly demanded. In this study, we present a scheme to identify gapmer antisense oligonucleotides (ASOs) targeting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA that efficiently inhibit viral replication. We synthesized approximately 300 gapmer ASOs designed to target various SARS-CoV-2 RNA regions and evaluated their activity in cell-based assays. Through a multistep screening in cell culture systems, we identified that ASO#41, targeting the coding region for viral main protease, reduced SARS-CoV-2 RNA levels in infected cells and inhibited virus-induced cytopathic effects. Antiviral effect of ASO#41 was also observed in iPS cell-derived human lung organoids. ASO#41 depleted intracellular viral RNAs during genome replication in an endogenous RNaseH-dependent manner. ASO#41 showed a wide range of antiviral activity against SARS-CoV-2 variants of concern including Alpha, Delta, and Omicron. Intranasal administration to mice exhibited intracellular accumulation of ASO#41 in the lung and significantly reduced the viral infectious titer, with milder body weight loss due to SARS-CoV-2 infection. Further chemical modification with phosphoryl guanidine-containing backbone linkages provided an elevation of anti-SARS-CoV-2 activity, with 23.4 nM of 50% antiviral inhibitory concentration, one of the strongest anti-SARS-CoV-2 ASOs reported so far. Our study presents an approach to identify active ASOs against SARS-CoV-2, which is potentially useful for establishing an antiviral strategy by targeting genome RNA of respiratory viruses.

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针对病毒 RNA 主要蛋白酶区的抗 SARS-CoV-2 gapmer 反义寡核苷酸。
鉴于新发/再发呼吸道病毒在全球范围内爆发的风险,建立新的抗病毒策略是非常必要的。在本研究中,我们提出了一种方案,以识别能有效抑制病毒复制的、靶向严重急性呼吸系统综合征冠状病毒 2(SARS-CoV-2)RNA 的间隙反义寡核苷酸(ASO)。我们合成了约 300 种针对不同 SARS-CoV-2 RNA 区域设计的间隙聚合物 ASO,并在基于细胞的试验中评估了它们的活性。通过在细胞培养系统中进行多步筛选,我们发现针对病毒主蛋白酶编码区的 ASO#41 能降低感染细胞中的 SARS-CoV-2 RNA 水平,并抑制病毒诱导的细胞病理效应。在 iPS 细胞衍生的人肺器官组织中也观察到了 ASO#41 的抗病毒作用。在基因组复制过程中,ASO#41 以内源性 RNaseH 依赖性方式消耗细胞内病毒 RNA。ASO#41 对包括 Alpha、Delta 和 Omicron 在内的 SARS-CoV-2 变体具有广泛的抗病毒活性。小鼠经鼻给药后,ASO#41 会在肺部细胞内蓄积,并显著降低病毒感染滴度,同时减轻 SARS-CoV-2 感染导致的体重减轻。通过含磷酸胍骨架连接的进一步化学修饰,提高了抗 SARS-CoV-2 的活性,50% 的抗病毒抑制浓度为 23.4 nM,是迄今报道的最强抗 SARS-CoV-2 ASO 之一。我们的研究提出了一种鉴别抗 SARS-CoV-2 活性 ASO 的方法,它可能有助于通过靶向呼吸道病毒的基因组 RNA 建立抗病毒策略。
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来源期刊
Antiviral research
Antiviral research 医学-病毒学
CiteScore
17.10
自引率
3.90%
发文量
157
审稿时长
34 days
期刊介绍: Antiviral Research is a journal that focuses on various aspects of controlling viral infections in both humans and animals. It is a platform for publishing research reports, short communications, review articles, and commentaries. The journal covers a wide range of topics including antiviral drugs, antibodies, and host-response modifiers. These topics encompass their synthesis, in vitro and in vivo testing, as well as mechanisms of action. Additionally, the journal also publishes studies on the development of new or improved vaccines against viral infections in humans. It delves into assessing the safety of drugs and vaccines, tracking the evolution of drug or vaccine-resistant viruses, and developing effective countermeasures. Another area of interest includes the identification and validation of new drug targets. The journal further explores laboratory animal models of viral diseases, investigates the pathogenesis of viral diseases, and examines the mechanisms by which viruses avoid host immune responses.
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