ABSTRACT
Faldaprevir analogue molecule (FAM) has been reported to effectively inhibit the catalytic activity of HCV NS3/4A protease, making it a potential lead compound against HCV. A series of HCV NS3/4A protease crystal structures were analyzed by bioinformatics methods, and the FAM-HCV NS3/4A protease crystal structure was chosen for this study. A 20.4 ns molecular dynamics simulation of the complex consists of HCV NS3/4A protease and FAM was conducted. The key amino acid residues for interaction and the binding driving force for the molecular recognition between the protease and FAM were identified from the hydrogen bonds and binding free energy analyses. With the driving force of hydrogen bonds and van der Waals, FAM specifically bind to the active pocket of HCV NS3/4A protease, including V130-S137, F152-D166, D77-D79 and V55, which agreed with the experimental data. The effect of R155K, D168E/V and V170T site-directed mutagenesis on FAM molecular recognition was analyzed for their effect on drug resistance, which provided the possible molecular explanation of FAM resistance. Finally, the system conformational change was explored by using free energy landscape and conformational cluster. The result showed four kinds of dominant conformation, which provides theoretical basis for subsequent design of Faldaprevir analogue inhibitors based on the structure of HCV NS3/4A protease.
Subject(s)
Antiviral Agents , Chemistry , Carrier Proteins , Chemistry , Drug Resistance, Viral , Endopeptidases , Hepacivirus , Molecular Dynamics Simulation , Mutagenesis, Site-Directed , Oligopeptides , Chemistry , Protease Inhibitors , Chemistry , Serine Proteases , Thiazoles , Chemistry , Viral Nonstructural Proteins , ChemistryABSTRACT
The progress of the second generation inhibitors of HCV NS3/4A serine protease, based on the modifications to the first generation ones (telaprevir, boceprevir, BILN-2061), is reviewed. The second generation inhibitors can significantly improve potency, pharmacokinetic profiles, and to some degree reduce the resistance induced by the first generation ones. The structure-activity relationships of these inhibitors are also discussed.
ABSTRACT
Objective:The aim of this study was to explore the vaccine potential of adenovirus transduced DC encoding for HCV nonstructural protein NS3 in a murine model.Methods:Dendritic cells were isolated from bone marrow of BALB/c mice and cultivated with cytokine cocktail for 7 days. Recombinant adenovirus was added to DC cultures at day 7,at a titration of 2?10~9 pfu/10~6 DC/well. FACS and Western blot techniques were used to identify the DC mature state and the protein expression, respectively. BALB/c mice were primed/boosted immunized with DC based vaccines at 3?10~5 cells for one mouse at a 10 day interval. 10 days after the second immunization, cytokines secretion and IFN-? generating T cells response were evaluated by ELISA and ELLISPOT analysis.Results:Our current data indicate that AdNS3 can stimulate the maturation of DC and efficiently express authentic HCV nonstructural proteins in infected cells. BALB/c mice inoculated with AdNS3 transduced DC twice generated strong IFN-? secreting T cells response(P