Small molecule inhibitors of the hepatitis C virus-encoded NS5A protein

Belda, Oscar; Targett-Adams, Paul

doi:10.1016/j.virusres.2012.09.007

Cited by 78 publications

(66 citation statements)

References 118 publications

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“…They used a luciferase-based reporter system and RTqPCR to measure viral protein translation and viral RNA copy numbers in replicon-harboring cells. They reported that DCV treatment leads to NS5A redistribution from the ER to LDs (25,37,45) and it is temporally correlated with suppression of viral replication (25). These results are consistent with our data in Fig.…”

Section: Discussionsupporting

confidence: 93%

Fast Hepatitis C Virus RNA Elimination and NS5A Redistribution by NS5A Inhibitors Studied by a Multiplex Assay Approach

Liu

Ndongwe

et al. 2015

Antimicrob Agents Chemother

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e While earlier therapeutic strategies for the treatment of hepatitis C virus (HCV) infection relied exclusively on interferon (IFN) and ribavirin (RBV), four direct-acting antiviral agents (DAAs) have now been approved, aiming for an interferon-free strategy with a short treatment duration and fewer side effects. To facilitate studies on the mechanism of action (MOA) and efficacy of DAAs, we established a multiplex assay approach, which employs flow cytometry, a Gaussia luciferase reporter system, Western blot analysis, reverse transcription-quantitative PCR (RT-qPCR), a limited dilution assay (50% tissue culture infectious dose [TCID 50 ]), and an image profiling assay that follows the NS5A redistribution in response to drug treatment. We used this approach to compare the relative potency of various DAAs and the kinetics of their antiviral effects as a potential preclinical measure of their potential clinical utility. We evaluated the NS5A inhibitors ledipasvir (LDV) and daclatasvir (DCV), the NS3/4A inhibitor danoprevir (DNV), and the NS5B inhibitor sofosbuvir (SOF). In terms of kinetics, our data demonstrate that the NS5A inhibitor LDV, followed closely by DCV, has the fastest effect on suppression of viral proteins and RNA and on redistribution of NS5A. In terms of MOA, LDV has a more pronounced effect than DCV on the viral replication, assembly, and infectivity of released virus. Our approach can be used to facilitate the study of the biological processes involved in HCV replication and help identify optimal drug combinations. Hepatitis C virus (HCV) infects approximately 3% of the world's population, which accounts for about 170 million chronically infected individuals. Annually, there are more than 350,000 deaths from HCV-related cirrhosis and hepatocellular carcinoma (1). In the United States, there are more than 3 million people with chronic HCV infection, and about 15,000 die from HCV-related liver disease each year.HCV is a positive-strand RNA virus grouped in the genus Hepacivirus within the family Flaviviridae (2). It is classified into at least 6 genotypes (gt), and its error-prone polymerase leads to more than 50 subtypes (3). The long open reading frame, which encodes the HCV polyprotein, is processed by host and viral proteases and gives rise to three structural proteins (the capsid protein core and envelope glycoproteins E1 and E2) and seven nonstructural (NS) proteins (p7, NS2, NS3, NS4A, NS4B, NS5A, and NS5B) (4). NS2 and p7 are essential for virus assembly but not RNA replication, whereas NS3 to NS5B are involved in a membrane-associated RNA replicase complex (RC) (5). The NS3 protein is composed of a serine protease and an RNA helicase/nucleoside triphosphatase (NTPase), NS4A serves as a cofactor for NS3 serine protease (6), NS5B is the RNA-dependent RNA polymerase (7), and NS5A is considered to play key roles in multiple steps of the HCV life cycle.NS5A is an ϳ450 amino acid phosphoprotein composed of an N-terminal amphipathic ␣-helix and three domains (domain I to domain III), each of wh...

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Section: Discussionsupporting

confidence: 93%

Fast Hepatitis C Virus RNA Elimination and NS5A Redistribution by NS5A Inhibitors Studied by a Multiplex Assay Approach

Liu

Ndongwe

et al. 2015

Antimicrob Agents Chemother

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“…Since the identification of DCV (formerly BMS-790052) as a potent, pangenotypic inhibitor of HCV RNA replication, 6 a number of studies have investigated the potential mechanism(s) of action of DCV and structurally related NS5A inhibitors. Collectively, these studies have identified several inhibitor properties that may explain their efficacy ( Figure 1B; reviewed in [7][8][9] ). First, their remarkable potency (picomolar to low nanomolar median effective concentration values) suggests that these inhibitors may synergistically disrupt multiple functions of NS5A in the HCV life cycle and/or target essential events in establishment of replication sites that in time will prevent continued HCV RNA replication.…”

Section: Hcv Ns5a Inhibitors Disrupt Replication Factory Formation: Amentioning

confidence: 99%

HCV NS5A Inhibitors Disrupt Replication Factory Formation: A Novel Mechanism of Antiviral Action

Eyre

Beard

2014

Gastroenterology

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After the embargo period via non-commercial hosting platforms such as their institutional repository  via commercial sites with which Elsevier has an agreement In all cases accepted manuscripts should: link to the formal publication via its DOI  bear a CC-BY-NC-ND license -this is easy to do, click here to find out how  if aggregated with other manuscripts, for example in a repository or other site, be shared in alignment with our hosting policy  not be added to or enhanced in any way to appear more like, or to substitute for, the published journal article Embargo ISSN Journal Name Embargo Period (months)0016-5085 Gastroenterology

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“…The initial lead compounds had only moderate potency, but subsequent medicinal chemistry efforts resulted in the discovery of drugs with unparalleled antiviral activity that are characterised by highly symmetrical dimeric structure. 3 The most studied of these 'palindromic'…”

Section: Raffaele De Francescomentioning

confidence: 99%

Daclatasvir: a team player rather than a prima donna in the treatment of hepatitis C

Aghemo

Francesco

2014

Gut

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Small molecule inhibitors of the hepatitis C virus-encoded NS5A protein

Cited by 78 publications

References 118 publications

Fast Hepatitis C Virus RNA Elimination and NS5A Redistribution by NS5A Inhibitors Studied by a Multiplex Assay Approach

Fast Hepatitis C Virus RNA Elimination and NS5A Redistribution by NS5A Inhibitors Studied by a Multiplex Assay Approach

HCV NS5A Inhibitors Disrupt Replication Factory Formation: A Novel Mechanism of Antiviral Action

Daclatasvir: a team player rather than a prima donna in the treatment of hepatitis C

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