Abstract：
The coronavirus disease 2019 (COVID-19) is caused by the etiological agent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). COVID-19, with the recurrent epidemics of new variants of SARS-CoV-2, remains a global public health problem, and new antivirals are still required. Some cholesterol derivatives, such as 25-hydroxycholesterol, are known to have antiviral activity against a wide range of enveloped and non-enveloped viruses, including SARS-CoV-2. At the entry step of SARS-CoV-2 infection, the viral envelope fuses with the host membrane dependent of viral spike (S) glycoproteins. From the screening of cholesterol derivatives, we found a new compound 26,27-dinorcholest-5-en-24-yne-3β,20-diol (Nat-20(S)-yne) that inhibited the SARS-CoV-2 S protein-dependent membrane fusion in a syncytium formation assay. Nat-20(S)-yne exhibited the inhibitory activities of SARS-CoV-2 pseudovirus entry and intact SARS-CoV-2 infection in a dose-dependent manner. Among the variants of SARS-CoV-2, inhibition of infection by Nat-20(S)-yne was stronger in delta and Wuhan strains, which predominantly invade into cells via fusion at the plasma membrane, than in omicron strains. The interaction between receptor-binding domain of S proteins and host receptor ACE2 was not affected by Nat-20(S)-yne. Unlike 25-hydroxycholesterol, which regulates various steps of cholesterol metabolism, Nat-20(S)-yne inhibited only de novo cholesterol biosynthesis. As a result, plasma membrane cholesterol content was substantially decreased in Nat-20(S)-yne-treated cells, leading to inhibition of SARS-CoV-2 infection. Nat-20(S)-yne having a new mechanism of action may be a potential therapeutic candidate for COVID-19.
摘要：
2019年冠状病毒病（COVID-19）是由病原体严重急性呼吸综合征冠状病毒2（SARS-CoV-2）引起的。随着SARS-CoV-2新变种的反复流行，COVID-19仍然是一个全球公共卫生问题，并且仍然需要新的抗病毒药物。一些胆固醇衍生物，如25-羟基胆固醇，已知对多种包膜和无包膜病毒具有抗病毒活性，包括SARS-CoV-2.在SARS-CoV-2感染的进入阶段，病毒包膜与依赖病毒刺突(S)糖蛋白的宿主膜融合。从胆固醇衍生物的筛选，我们发现了一种新的化合物26,27-dinorcholest-5-en-24-yne-3β，在合胞体形成测定中抑制SARS-CoV-2S蛋白依赖性膜融合的20-二醇(Nat-20(S)-yne)。Nat-20（S）-yne以剂量依赖的方式表现出SARS-CoV-2假病毒进入和完整的SARS-CoV-2感染的抑制活性。在SARS-CoV-2的变种中，三角洲和武汉菌株对Nat-20（S）-yne感染的抑制作用更强，主要通过质膜融合侵入细胞，而不是omicron菌株。S蛋白的受体结合域与宿主受体ACE2之间的相互作用不受Nat-20（S）-yne的影响。不像25-羟基胆固醇,调节胆固醇代谢的各个步骤，Nat-20(S)-炔仅抑制从头胆固醇生物合成。因此,Nat-20(S)-yne处理细胞的质膜胆固醇含量显著降低,导致抑制SARS-CoV-2感染。具有新作用机制的Nat-20(S)-yne可能是COVID-19的潜在治疗候选药物。
