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Then, we screened these chemical drugs by in vitro fluorescence resonance energy transfer (FRET) enzymatic assays at a single concentration (100 μM Fig. So, we selected a cluster of 18 chemical drugs that were designed to target the different viral proteases and proteasome (Table 1).
GRAPHPAD PRISM 8 DRUG TEST PRO
At the same time, we found that the M pro of SARS-CoV-2 has high homology with other CoV M pro (Supplementary Fig. In the beginning, we obtained soluble and pure M pro protein of the SARS-CoV-2 with an extra glycine residue at the N-terminus (GM pro) after TEV cleavage by expression in E. Taken together, these data provide critical information for the optimization and design of more potent inhibitors against the emerging pathogen SARS-CoV-2. Additionally, the high-resolution crystal structures of SARS-CoV-2 M pro complex with these two inhibitors are solved to figure out their mechanism of inhibition. Moreover, combination of GC376 with Remdesivir treatment can enhance antiviral activity. Here we show both Boceprevir and GC376 can inhibit M pro activity and SARS-CoV-2 in Vero cells. As there is neither specific antiviral agents nor available vaccines, repurposing of clinically approved drugs to combat the COVID-19 is urgently needed. Actually, viral proteases are also promising targets for many different viruses including hepatitis C virus (HCV)and human immunodeficiency virus (HIV) 23. The coronavirus main protease (M pro, 3CLpro) is essential for viral polyproteins processing and maturation, therefore, it is recognized as an attractive drug target. Therefore, it is urgent to develop specific drugs against the virus.
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However, to date, no clinically approved specific drugs or vaccines are available to treat the disease. Some old drugs such as Remdesivir, Favipiravir, and Chloroquine/Hydroxychloroquine, and also traditional Chinese medicines showed potential for the treatment of COVID-19 19– 22. Recent studies have shown that the SARS-CoV-2 uses the same Angiotensin-converting enzyme 2 (ACE2) receptor as SARS-CoV 4, and the structural basis of receptor recognition was quickly elucidated to provide important basis for the molecular understanding of virus entry process and the development of potential antiviral inhibitors 15– 18. Sequence comparison has showed that the SARS-CoV-2 has the closest relationship (96.2%) with the bat SARS-like coronavirus 4, 12, but the origin of this virus was yet to be identified. The patients with older age and comorbidities such as hypertension, diabetes, and coronary heart disease will have high risk of death 14. Most of the patients have a good prognosis, and the symptoms of children are relatively mild 14. Some mildly ill patients do not have fever and obvious respiratory symptoms 14. The longest observed duration of viral shedding in survivors was 37 days 14. The patient at incubation period is contagious, and the median duration of viral shedding was 20 days in survivors, but the SARS-CoV-2 was detectable until death in non-survivors 14. The median incubation period of the disease is 4 days, and the longest is no more than 41 days 13. The clinical manifestations of COVID-19 include fever, fatigue, dry cough, headache, and diarrhea 13. The other six coronaviruses are the low-pathogenicity members including HCoV-OC43, HCoV-HKU1, HCoV-NL63, and HCoV-229E, and highly pathogenic SARS-CoV and MERS-CoV 12. Human coronavirus 2019 (HCoV-19) 10 is the seventh coronavirus capable of infecting humans, which was later named SARS-CoV-2 by the International Committee on Taxonomy of Viruses (ICTV) 11 though with different name being proposed. As of 28 March, the disease has caused more than 500,000 human infections with over 20,000 deaths globally 9. The World Health Organization (WHO) named the infectious disease as COVID-19, and declared a global pandemic on 11 March 2020 8. The virus has rapidly spread to more than 200 countries in the world 6, 7. In December 2019, a novel coronavirus was discovered due to emerging viral pneumonia cases 1– 5.