热毒宁注射液有效成分对SARS-CoV-2突变株保持高效抑制力

XIAO Zhen; XU Huan; QU Ze-yang; MA Xin-yuan; HUANG Bo-xuan; SUN Meng-si; WANG Bu-qing; WANG Guan-yu

doi:10.1007/s11655-022-3686-5

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热毒宁注射液有效成分对SARS-CoV-2突变株保持高效抑制力

Original Article | Updated：2023-02-22

- 热毒宁注射液有效成分对SARS-CoV-2突变株保持高效抑制力
- Active Ingredients of Reduning Injection Maintain High Potency against SARS-CoV-2 Variants
- 中国结合医学杂志（英文版） 2023年29卷第3期页码：205-212
- Affiliations：
  
  1.Department of Biology, School of Life Sciences, Southern University of Science and Technology, Shenzhen, Guangdong(518055), China
  2.Guangdong Provincial Key Laboratory of Computational Science and Material Design, Shenzhen, Guangdong Province (518055), China
  3.Institute of Chemical Biology, Shenzhen Bay Laboratories, Shenzhen, Guangdong Province (518132), China
  4.State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing(100191), China
  5.Institute for Advanced Study, Shenzhen University, Shenzhen, Guangdong Province(518052), China
  6.Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu Province (214122), China
  7.School of Medicine Life and Health Sciences, Chinese University of Hong Kong, Shenzhen, Guangdong Province (518172), China
  8.Center for Endocrinology and Metabolic Diseases, Second Affiliated Hospital, The Chinese University of Hong Kong, Shenzhen, Guangdong Province (518172), China
- Author bio：
  
  Prof. WANG Guan-yu, E-mail: wangguanyu@cuhk.edu.cn
- Funds：
  
  National Key R&D Plan Project(2019YFA0906002);National Natural Science Foundation of China(32070681)
- DOI：10.1007/s11655-022-3686-5
  中图分类号：
- 纸质出版日期：2023-03，
  
  网络出版日期：2022-11-14，
  
  录用日期：2022-08-05
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热毒宁注射液有效成分对SARS-CoV-2突变株保持高效抑制力[J]. 中国结合医学杂志（英文版）, 2023,29(3):205-212.

XIAO Zhen, XU Huan, QU Ze-yang, et al. Active Ingredients of Reduning Injection Maintain High Potency against SARS-CoV-2 Variants[J]. Chinese Journal of Integrative Medicine, 2023,29(3):205-212.
热毒宁注射液有效成分对SARS-CoV-2突变株保持高效抑制力[J]. 中国结合医学杂志（英文版）, 2023,29(3):205-212. DOI： 10.1007/s11655-022-3686-5.

XIAO Zhen, XU Huan, QU Ze-yang, et al. Active Ingredients of Reduning Injection Maintain High Potency against SARS-CoV-2 Variants[J]. Chinese Journal of Integrative Medicine, 2023,29(3):205-212. DOI： 10.1007/s11655-022-3686-5.

摘要

目的:

探讨热毒宁注射液中槲皮素和木犀草素两种成分的抗冠状病毒作用及相应机制.

方法:

设计假病毒系统

测试槲皮素和木犀草素对SARS-CoV-2感染的抑制作用及其相应的细胞毒性. 测试了木犀草素对SARS-CoV-2及其突变株假病毒的活性. 通过Autodock Vina 1.1.230和PyMol进行虚拟筛选以预测结合位点. 为了验证对接结果

使用表面等离子体共振 (SPR) 检测化合物与冠状病毒不同蛋白的结合亲和力. 通过对感染了HCoV-OC43的横纹肌肉瘤细胞进行间接免疫荧光测定

进一步测试了槲皮素和木犀草素对其他冠状病毒的抑制作用.

结果:

木犀草素和槲皮素对SARS-CoV-2假病毒的抑制呈强烈的剂量依赖性

分别为8.817和52.98μmol/L. 它们对BHK21-hACE2的细胞毒性分别为177.6和405.1μmol/L. 此外

木犀草素显著阻断了4种SARS-CoV-2变体假病毒的进入

均低于7μmol/L. 虚拟筛选和SPR证实木犀草素与S蛋白结合

槲皮素与3CLpro、PLpro和解旋酶蛋白的活性中心结合. 槲皮素和木犀草素对HCoV-OC43的抑制率超过99%.

结论:

本研究揭示了槲皮素和木犀草素抑制 SARS-CoV-2及其突变株的感染机制. 热毒宁注射液是很有前景的COVID-19治疗药物.

Abstract

Objective:

To investigate the anti-coronavirus potential and the corresponding mechanisms of the two ingredients of Reduning Injection: quercetin and luteolin.

Methods:

A pseudovirus system was designed to test the efficacy of quercetin and luteolin to inhibit SARS-CoV-2 infection and the corresponding cellular toxicity. Luteolin was tested for its activities against the pseudoviruses of SARS-CoV-2 and its variants. Virtual screening was performed to predict the binding sites by Autodock Vina 1.1.230 and PyMol. To validate docking results

surface plasmon resonance (SPR) was used to measure the binding affinity of the compounds with various proteins of the coronaviruses. Quercetin and luteolin were further tested for their inhibitory effects on other coronaviruses by indirect immunofluorescence assay on rhabdomyosarcoma cells infected with HCoV-OC43.

Results:

The inhibition of SARS-CoV-2 pseudovirus by luteolin and quercetin were strongly dose-dependent

with concentration for 50% of maximal effect (EC

) of 8.817 and 52.98 μmol/L

respectively. Their cytotoxicity to BHK21-hACE2 were 177.6 and 405.1 μmol/L

respectively. In addition

luetolin significantly blocked the entry of 4 pseudoviruses of SARS-CoV-2 variants

with EC

lower than 7 μmol/L. Virtual screening and SPR confirmed that luteolin binds to the S-proteins and quercetin binds to the active center of the 3CLpro

PLpro

and helicase proteins. Quercetin and luteolin showed over 99% inhibition against HCoV-OC43.

Conclusions:

The mechanisms were revealed of quercetin and luteolin inhibiting the infection of SARS-CoV-2 and its variants. Reduning Injection is a promising drug for COVID-19.

关键词

热毒宁注射液中药木犀草素槲皮素SARS-CoV-2

Keywords

Reduning injectionChinese medicineluteolinquercetinSARS-CoV-2

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