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Objective: To synthesize the enterovirus 71 3C protease inhibitor NK-1.8k and optimize the synthetic process. Methods: With N-Boc-L-glutamic acid dimethyl ester as the starting material, the target compound NK-1.8k was synthesized via the substitution, reductive amination, deprotection, amide condensation, hydrolysis, and reduction reactions. Compared with the original synthetic route, the tandem string type in the important intermediate 5 synthesis was changed to parallel type, thereby the total synthetic reactions were condensed from seven steps to six steps, and some post-processing methods were also optimized. Results: The structures of intermediates and the target compound were confirmed by MS and 1H NMR data, and the total yield of the target compound synthesis was increased to 13.3% from 10.7% of the original route. Conclusion: The synthetic route established in this article for NK-1.8k is reasonable and feasible, the raw materials used are cheap and easily available, the operation is simple, most of the reaction conditions are mild and controllable, the post-processing is simple, the intermediates are easy to separate and purify, the steps are short, and the yield is high. This method provides a valuable reference for the further synthesis of NK-1.8k and similar products.
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Objective: To search for Chinese herbal monomer with the antiviral activity of enterovirus 71 (EV71) 3C protease by virtual screening. Methods: A total of 1998 monomers from antiviral Chinese herbs downloaded from TCM Database@Taiwan database were chosen as screening ligands, and EV71 3C protease was selected as a target. The binding mode of Chinese herbal monomer and EV71 3C protease was simulated through the molecular docking tools of AutoDock Vina. The monomers were sorted according to the binding free energy. The binding poses of the monomers with docking scores less than -37.673 kJ/mol were visualized by Discovery Studio 2018 Visualizer program. Results: Two monomers, namely, procyanidin B5 and 1,2,3,6-tetra-O-galloyl- β-D-glucose, were obtained with the lowest binding energies. Conclusion: The promising candidate monomers with anti-EV71 3C protease activity were screened out from traditional Chinese medicine, providing an alternative method for further development of therapeutically useful anti-EV71 agents.
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Picornaviruses(PV)and coronaviruses(CoV) are positive-stranded RNA viruses. Pathogens in the family can cause hand,foot and mouth disease,myocarditis, common cold ,severe respiratory and intestinal diseases. 3C and 3CL proteases, belonging to cysteine proteases,are required to process polyproteins into mature proteins for viral replication,which plays an impor?tant role in viral replication because substrate binding sites are highly conservative and have similar catalytic mechanism. 3C and 3CL proteases are different from protease in the human body ,which represents a promising anti-viral drug target. Using 3C and 3CL pro?teinase structural similarities,broad spectrum protease inhibitors have been found successfully. This review describes recent develop?ments of broad spectrum protease inhibitors targeting on 3C and 3CL proteases,and briefly illustrates the mechanism of the inhibitors, which may benefit to the development of virus therapy.
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Picornaviruses (PV) and coronaviruses (CoV) are positive-stranded RNA viruses. Pathogens in the family can cause hand, foot and mouth disease, myocarditis, common cold, severe respiratory and intestinal diseases. 3C and 3CL proteases, belonging to cysteine proteases, are required to process polyproteins into mature proteins for viral replication, which plays an important role in viral replication because substrate binding sites are highly conservative and have similar catalytic mechanism. 3C and 3CL proteases are different from protease in the human body, which represents a promising anti-viral drug target. Using 3C and 3CL proteinase structural similarities, broad spectrum protease inhibitors have been found successfully. This review describes recent developments of broad spectrum protease inhibitors targeting on 3C and 3CL proteases, and briefly illustrates the mechanism of the inhibitors, which may benefit to the development of virus therapy.
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Objective:To explore novel compounds with anti-EV71 activity. Methods: A series of oligopeptides were designed based on the active groove of the EV71 3C protease using molecular simulation technology. Furthermore, some oligopeptides were syn-thesized and tested for the inhibition ability against EV71 in vitro. Results:Pentapeptides had higher binding affinity to the EV71 3C protease than the other oligopeptides, such as compounds 25 and 16. Therefore, compounds 25 and 16 were synthesized and tested in vitro for the anti-EV71 activity, and compound 25 exhibited potent anti-EV71 activity(EC50 =1. 36 μmol·L-1, CC50 =211. 94 μmol ·L-1, SI=155. 84), while compound 16 had no evident effect on EV71. Conclusion: It is worthy of further investigation on com-pound 25 as a novel scaffold of anti-EV71inhibitor.
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Objective To obtain a novel tool-enzyme for genetic engineering with good solubility,strong specificity of enzyme digestion and maintaining the enzyme activity at low temperature by using E.coli expression system to express self-processed re-combinant MBP-HRV 3C fusion protease.Methods The cDNA encoding HRV 3C protease was cloned into pRSF-Duet vector.The recombinant plasmid was transferred into E.coli BL21 (DE3)for expression.HRV 3C protease was obtained through Nichol col-umn affinity purification.The cleavage activity of HRV 3C protease was determined by in vivo experiment.Results HRV 3C prote-ase was highly expressed in E.coli expression system,and the obtained HRV 3C protease could recognize and digest HRV 3C site. Conclusion A novel tool-enzyme for genetic engineering is obtained.