Cellular Growth Kinetics Distinguish a Cyclophilin Inhibitor from an HSP90 Inhibitor as a Selective Inhibitor of Hepatitis C Virus

Beran, Rudolf K. F.; Sharma, Rachna; Corsa, Amoreena C.; Yang, Tian; Golde, Justin; Lundgaard, Greta L.; Delaney, William E.; Zhong, Weidong; Greenstein, Andrew E.

doi:10.1371/journal.pone.0030286

Cited by 16 publications

(11 citation statements)

References 35 publications

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“…Initial attempts to establish long-term HCV persistently-infected cell cultures failed because the majority of the infected cells died after reaching confluence (data not shown). Thus, based on previously described methods [28], [29], [31], we added DMSO to 1% final concentration to Huh7-Lunet CD81 cells growing on 12-well plates. Within 3–4 days, a confluent cellular layer formed in each well and we infected the cultures using high-titer genotype 2a HCV or genotype 1b/2a HCV stocks.…”

Section: Resultsmentioning

confidence: 99%

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Hepatitis C Viral Entry Inhibitors Prolong Viral Suppression by Replication Inhibitors in Persistently-Infected Huh7 Cultures

et al. 2013

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Efforts to treat HCV patients are focused on developing antiviral combinations that lead to the eradication of infection. Thus, it is important to identify optimal combinations from the various viral inhibitor classes. Based on viral dynamic models, HCV entry inhibitors are predicted to reduce viral load in a monophasic manner reflecting the slow death rate of infected hepatocytes (t1/2 = 2–70 days) and the protection of naïve, un-infected cells from HCV infection. In contrast, replication inhibitors are predicted to reduce viral load in a biphasic manner. The initial rapid reduction phase is due to the inhibition of virus production and elimination of plasma virus (t1/2∼3 hours). The second, slower reduction phase results from the elimination of infected hepatocytes. Here we sought to compare the ability of HCV entry and replication inhibitors as well as combinations thereof to reduce HCV infection in persistently-infected Huh7 cells. Treatment with 5×EC50 of entry inhibitors anti-CD81 Ab or EI-1 resulted in modest (≤1 log10 RNA copies/ml), monophasic declines in viral levels during 3 weeks of treatment. In contrast, treatment with 5×EC50 of the replication inhibitors BILN-2016 or BMS-790052 reduced extracellular virus levels more potently (∼2 log10 RNA copies/ml) over time in a biphasic manner. However, this was followed by a slow rise to steady-state virus levels due to the emergence of resistance mutations. Combining an entry inhibitor with a replication inhibitor did not substantially enhance the rate of virus reduction. However, entry/replication inhibitor and replication/replication inhibitor combinations reduced viral levels further than monotherapies (up to 3 log10 RNA copies/ml) and prolonged this reduction relative to monotherapies. Our results demonstrated that HCV entry inhibitors combined with replication inhibitors can prolong antiviral suppression, likely due to the delay of viral resistance emergence.

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Section: Resultsmentioning

confidence: 99%

“…HCV persistently-infected cultures were established using methods adapted from Sainz and Chisari (2006) [28] and Beran et al ., (2012) [29]. Briefly, Huh7-Lunet-CD81 cells [26] were seeded in 12-well plates at a density of 50,000 cells/well.…”

Section: Methodsmentioning

confidence: 99%

Hepatitis C Viral Entry Inhibitors Prolong Viral Suppression by Replication Inhibitors in Persistently-Infected Huh7 Cultures

et al. 2013

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“…Recently, experimental data on the antiviral activity of gemcitabine have been widely reported (Shin et al, 2018;Ianevski et al, 2019). The broad range of viruses against which gemcitabine has shown activity includes HIV-1 (Rawson et al, 2013;Clouser et al, 2010Clouser et al, , 2012, murine leukaemia virus (MuLV) (Clouser et al, 2012), HIV-2 (Beach et al, 2014), ZIKV (Kuivanen et al, 2017), enteroviruses such as poliovirus (Kang et al, 2015;Zhang et al, 2017), rhinoviruses (Song et al, 2017), HCV (Beran et al, 2012), coronaviruses (Dyall et al, 2014), Sindbis virus (SINV), HSV-1 and FLUAV (Denisova et al, 2012). The synergistic effect of gemcitabine combined with low doses of other nucleoside analogues was also reported.…”

Section: Nucleoside Antimetabolites -Approved Antineoplastic Drugs Wimentioning

confidence: 99%

“…Myelodysplastic syndrome (2004) AdV (Alexeeva et al (2001); (Alexeeva et al, 2015) HMPV (Bösl et al, 2019) HIV-1 (Dapp et al, 2009;Rawson et al, 2016a) HIV-1 -synergy in combination with resveratrol (Rawson et al, 2016b) HIV-2 (Beach et al, 2014) RVFV ( (Clouser et al, 2012) HIV-1 -synergy in combination with decitabine (Clouser et al, 2010) MuLV (Clouser et al, 2012) HIV-2 (Beach et al, 2014) ZIKV (Kuivanen et al, 2017) Enteroviruses including poliovirus (Kang et al, 2015;Zhang et al, 2017) Enteroviruses -synergy with ribavirin (Kang et al, 2015) Rhinoviruses (Song et al, 2017) HCV (Beran et al, 2012) Coronaviruses (Dyall et al, 2014) SINV, HSV-1, FLUAV (Denisova et al, 2012) Gram-positive bacteria (Jordheim et al, 2012;Sandrini et al, 2007;…”

Section: Introductionmentioning

confidence: 99%

Selected nucleos(t)ide-based prescribed drugs and their multi-target activity

Pastuch-Gawołek

Gillner

Król

et al. 2019

European Journal of Pharmacology

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A B S T R A C T Nucleos(t)ide analogues play pivotal roles as antiviral, cytotoxic or immunosuppressive agents. Here, we review recent reports of nucleoside analogues that exhibit broad-spectrum activity towards multiple life-threatening RNA and DNA viruses. We also present a discussion about nucleoside antimetabolites-approved antineoplastic agents-that have recently been shown to have antiviral and/or antibacterial activity. The approved drugs and drug combinations, as well as recently identified candidates for investigation and/or experimentation, are discussed. Several examples of repurposed drugs that have already been approved for use are presented. This strategy can be crucial for the first-line treatment of acute infections or coinfections and for the management of drug-resistant strains.T against viral infections of great medical importance (e.g., herpes simplex virus (HSV), varicella zoster virus (VZV), human immunodeficiency virus (HIV), hepatitis B virus (HBV) and hepatitis C virus (HCV)) and/or that are known antineoplastic agents used to treat cancer. Approved nucleos(t)ide-based drugs with multi-target activity and compounds being tested in clinical trials or evaluated in preclinical assays are listed in Table 1. In this paper, the term "approval date" means the date on which the drug was approved by the US Food and Drug Administration (FDA) unless otherwise stated. G. Pastuch-Gawołek, et al. Multi-target nucleos(t)ide-based drugs in the treatment and prophylaxis of HIV, HBV and HCV infections

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“…HCV persistently infected cultures were established using previously described methods (5,31,32). Briefly, Huh7-Lunet-CD81 cells (22) were seeded in 12-well plates at a density of 50,000 cells/well.…”

Section: Methodsmentioning

confidence: 99%

A Small-Molecule Inhibitor of Hepatitis C Virus Infectivity

Bush

Pokrovskii

Saito

et al. 2014

Antimicrob Agents Chemother

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One of the most challenging goals of hepatitis C virus (HCV) research is to develop well-tolerated regimens with high cure rates across a variety of patient populations. Such a regimen will likely require a combination of at least two distinct direct-acting antivirals (DAAs). Combining two or more DAAs with different resistance profiles increases the number of mutations required for viral breakthrough. Currently, most DAAs inhibit HCV replication. We recently reported that the combination of two distinct classes of HCV inhibitors, entry inhibitors and replication inhibitors, prolonged reductions in extracellular HCV in persistently infected cells. We therefore sought to identify new inhibitors targeting aspects of the HCV replication cycle other than RNA replication. We report here the discovery of the first small-molecule HCV infectivity inhibitor, GS-563253, also called HCV infectivity inhibitor 1 (HCV II-1). HCV II-1 is a substituted tetrahydroquinoline that selectively inhibits genotype 1 and 2 HCVs with low-nanomolar 50% effective concentrations. It was identified through a high-throughput screen and subsequent chemical optimization. HCV II-1 only permits the production and release of noninfectious HCV particles from cells. Moreover, infectious HCV is rapidly inactivated in its presence. HCV II-1 resistance mutations map to HCV E2. In addition, HCV-II prevents HCV endosomal fusion, suggesting that it either locks the viral envelope in its prefusion state or promotes a viral envelope conformation change incapable of fusion. Importantly, the discovery of HCV II-1 opens up a new class of HCV inhibitors that prolong viral suppression by HCV replication inhibitors in persistently infected cell cultures.O ne of the most challenging goals of hepatitis C virus (HCV) research is to develop well-tolerated regimens with high cure rates. The standard of care for HCV patients over the past decade has been to treat with pegylated interferon combined with ribavirin. Relatively recently, the HCV NS3-4A protease inhibitors teleprevir and boceprevir were added to this standard of care and have improved the sustained virologic response (1). However, poor tolerability to treatments containing pegylated interferon has motivated researchers to attempt to develop a variety of other interferon-free treatments (2). An interferon-free cure will likely require a combination of at least two antivirals with differing modes of action (3, 4). Combining multiple antivirals with different resistance profiles increases the number of resistance mutations required for viral breakthrough (5). Studies to determine optimal antiviral combinations which take into account the probability of emergence of specific resistance mutations as well as the subsequent viral fitness (3, 6, 7) are under way.We recently demonstrated that the in vitro combination of HCV entry inhibitors with HCV replication inhibitors could prolong the efficacy compared to a single treatment with either inhibitor class. Specifically, we studied the entry inhibitor anti-CD81 anti...

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Cellular Growth Kinetics Distinguish a Cyclophilin Inhibitor from an HSP90 Inhibitor as a Selective Inhibitor of Hepatitis C Virus

Cited by 16 publications

References 35 publications

Hepatitis C Viral Entry Inhibitors Prolong Viral Suppression by Replication Inhibitors in Persistently-Infected Huh7 Cultures

Hepatitis C Viral Entry Inhibitors Prolong Viral Suppression by Replication Inhibitors in Persistently-Infected Huh7 Cultures

Selected nucleos(t)ide-based prescribed drugs and their multi-target activity

A Small-Molecule Inhibitor of Hepatitis C Virus Infectivity

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