Please wait a minute...

中国生物工程杂志

China Biotechnology
China Biotechnology  2013, Vol. 33 Issue (12): 51-56    DOI:
    
Establishment and Application of a System for Drug Screening Targeting MERS-CoV Main Proteinase
ZHANG Ning1,3, PAN Li1,3, NIU Guo-jun3, WANG Wen-ming4, ZHOU Hong-gang2, YANG Cheng2,3
1. College of Biological Engineering, Tianjin University of Science & Technology, Tianjin 300457, China;
2. College of Pharmacy, Nankai University, Tianjin 300457, China;
3. Tianjin International Joint Academy of Biomedicine Co.Ltd., Tianjin 300457, China;
4. College of Life Sciences, Nankai University, Tianjin 300457, China
Download: HTML   PDF(766KB) HTML
Export: BibTeX | EndNote (RIS)      

Abstract  Objective: Targeting MERS-CoV main proteinase NSP5, a new system for drug screening was established for inhibitors screening by using fluorescence resonance energy transfer (FRET). Methods: Target protein was obtained using prokaryotic expression system and technologies of gene recombination, protein-expression and purification. Proteinase activity of target protein was examined by FRET assays. Finally, a system for drug screening was established and optimized before abundant compounds screening. Results: Using the drug screening system, eight compounds with high inhibition ratio were selected. Notably, the IC50 of MDCCCL002330 is minimal, namely 0.43μmol/L. Conclusion: The established system targeting MERS-CoV main proteinase NSP5 is suitable for inhibitors screening, which can be used to promote research and development of lead compounds.

Key wordsNSP5      Inhibitors      High Throughput Screening      MERS-CoV     
Received: 17 September 2013      Published: 25 December 2013
ZTFLH:  Q819  
Cite this article:

ZHANG Ning, PAN Li, NIU Guo-jun, WANG Wen-ming, ZHOU Hong-gang, YANG Cheng. Establishment and Application of a System for Drug Screening Targeting MERS-CoV Main Proteinase. China Biotechnology, 2013, 33(12): 51-56.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2013/V33/I12/51

[1] Zaki A M, van Boheemen S, Bestebroer T M, et al. Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia. N Engl J Med, 2012, 367(19):1814-1820.
[2] Bermingham A, Chand M A, Brown C S, et al. Severe respiratory illness caused by a novel coronavirus, in a patient transferred to the United Kingdom from the Middle East, September 2012.Euro Surveill, 2012, 17(40):20290.
[3] World Health Organization. Novel coronavirus infection-update. http://www.who.int/csr/disease/coronavirus_infections/update_20130813/en/index.html.
[4] Raoul J de Groot, Susan C Baker, Ralph S Baric, et al. Middle East Respiratory Syndrome Coronavirus (MERS-CoV): Announcement of the coronavirus study group. Journal of Virology, 2013, 87(14):7790-7792.
[5] van Boheemen S, de Graaf M, Lauber C, et al. Genomic characterization of a newly discovered coronavirus associated with acute respiratory distress syndrome in humans. MBio, 2012, 3(6): e00473-12.
[6] Lu G, Liu D. SARS-like virus in the Middle East: a truly bat-related coronavirus causing human diseases. Protein Cell, 2012, 3(11):803-805.
[7] Memish A Ziad, Nischay Mishra, Kevin J, et al. Middle East Respiratory Syndrome Coronavius in Bats, Saudi Arabia. Emerging Infectious Diseases, 2013, (in press).
[8] Reusken C B, Haagmans B L, MÜller M A, et al. Middle East respiratory syndrome coronavirus neutralizing serum antibodies in dromedary camels: a comparative serological study. The Lancet Infect Diseases, 2013, (in press).
[9] Raj V S, Mou H, Smits S L, et al. Dipeptidyl peptidase 4 is a functional receptor for the emerging human coronavirus-EMC. Nature, 2013, 495(7440):251-254.
[10] Lu G, Hu Y, Wang Q, et al. Molecular basis of binding between novel human coronavirus MERS-CoV and its receptor CD26. Nature, 2013, 500(7461):227-231.
[11] Yang H, Bartlam M, Rao Z. Drug design targeting the main protease, the Achilles heel of coronaviruses. Curr Pharm Des. 2006, 12(35):4573-4590.
[12] Ren Z, Yan L, Zhang N, et al. The newly emerged SARS-like coronavirus HCoV-EMC also has an Achilles' heel": current effective inhibitor targeting a 3C-like protease. Protein Cell. 2013, 4(4):248-250.
[13] Ziebuhr J, Heusipp G, Siddell S G. Biosynthesis, purification and characterization of the human coronavirus 229E 3C-like proteinase. J Virol, 1997, 71(5):3992-3997.
[14] Barnard D L, Kumaki Y. Recent developments in anti-severe acute respiratory syndrome coronavirus chemotherapy. Future Virol, 2011, 6(5):615-631.
[15] Thanigaimalai P, Konno S, Yamamoto T, et al. Development of potent dipeptide-type SARS-CoV 3CL protease inhibitors with novel P3 scaffolds: Design, synthesis, biological evaluation, and docking studies. Eur J Med Chem, 2013, (in press).
[16] Zhao Q, Weber E, Yang H. Recent developments on coronavirus main protease/3C like protease inhibitors. Recent Pat Anbiinfect Drug Discovery, 2013, 8(2):150-156.
[17] Blanchard J E, Elowe N H, Huitema C, et al. High-throughput screening identifies inhibitors of the SARS coronavirus main proteinase. Chem Biol, 2004, 11(10):1445-1453.
[1] LI Wen,CHEN Jie,HU Wei-nan,QI Ya-yun,FU Yi-hong,LIU Jia-min,WANG Zhen-chao,OUYANG Gui-ping. Research Advances in the Study of EGFR Mutations Resistance and Its Small Molecule Inhibitors[J]. China Biotechnology, 2019, 39(10): 97-104.
[2] HE Lin-wei, LIU Zhang-min, FENG Yan, CUI Li. High Throughput Screening Method and Application for L-glutamate Specific Aminotransferase[J]. China Biotechnology, 2017, 37(8): 59-65.
[3] Li-na GU,Liang-zhi LI,Wei-qiang GUO,Jing-sheng GU,Xue-mei YAO,Xin JU. The Regulation on Polyols Production by Trichosporonoides oedocephalis with HOG1 Inhibitors and Its Mechanism[J]. China Biotechnology, 2017, 37(12): 40-48.
[4] GUO Xue-jiao, ZHA Jian, YAO Kun, WANG Xin, LI Bing-zhi, YUAN Ying-jin. Accelerated Ethanol Production by a Tolerant Saccharomyces cerevisiae to Inhibitor Mixture of Furfural, Acetic Acid and Phenol[J]. China Biotechnology, 2016, 36(5): 97-105.
[5] QIN Ling-yun, CHEN Rong, SU Zheng-ding. Design of Mdm2/MdmX Inhibitors[J]. China Biotechnology, 2015, 35(9): 78-84.
[6] TIAN Cong-hui, TANG Yan-ting, WANG Quan, ZHOU Hong-gang. Estabilishment and Application of a Model for Drug Screening Targeting Neprilysin Proteinase[J]. China Biotechnology, 2015, 35(2): 52-58.
[7] ZHANG Dong-xu. Recent Advances in Biological Detoxification of Inhibitors in Lignocellulose Hydrolysate[J]. China Biotechnology, 2013, 33(5): 120-124.
[8] NIE Lun, WU Wen-yan. RANTES Derivates and HIV-1 Entry Inhibitor[J]. China Biotechnology, 2013, 33(2): 96-102.
[9] WANG Jian-hua, QUAN Chun-shan, ZHAO Peng-chao, FAN Sheng-di. Study on the Inhibition Effect of DKP on the Biofilms Formed by Three Pathogens[J]. China Biotechnology, 2011, 31(8): 61-65.
[10] LV Yi, ZHENG Jin-ping. The Inhibitory Effect of Arresten Protein on Angiogesis[J]. China Biotechnology, 2011, 31(01): 19-23.
[11] WEN Lei, SONG Na-Ling, HE Xin, DIAO Qi-Ren. Type Ⅳ Collagen-derived Angiogenesis Inhibitors[J]. China Biotechnology, 2010, 30(05): 116-121.
[12] DAN Liu, JIANG E-Qin, BAI Yan-Qiu. Establishment of High Throughput Screening System of mGluR5[J]. China Biotechnology, 2010, 30(05): 81-86.
[13] YANG Lian-Zhi- Li-Chu-Fang- Lin-Zuo-Xian- Sun-Cai-Jun- Chen-Ling. Expression and application of neuraminidase of influenza virus[J]. China Biotechnology, 2009, 29(05): 6-10.
[14] . Establishment of a Cell Based High Throughput Screening Model for Discovering New Agonists of Histamine H3 Receptor[J]. China Biotechnology, 2008, 28(9): 61-67.
[15] Xiujun WANG . Advances in Mechanism of Herbicide in Inhibiting Amino Acid Biosynthesis and Herbicide-tolerant Transgenic Plants[J]. China Biotechnology, 2008, 28(2): 110-116.