A screen for novel hepatitis C virus RdRp inhibitor identifies a broad-spectrum antiviral compound

ABSTRACT To address the need for broad-spectrum antiviral activity characterization of hepatitis C virus (HCV) polymerase inhibitors, we created a panel of intergenotypic chimeric replicons containing nonstructural (NS) protein NS5B sequences from genotype 2b (GT2b), GT3a, GT4a, GT5a, and GT6a HCV isolates. Viral RNA extracted from non-GT1 HCV patient plasma was subjected to reverse transcription. The NS5B region was amplified by nested PCR and introduced into the corresponding region of the GT1b (Con-1) subgenomic reporter replicon by Splicing by Overlap Extension (SOEing) PCR. Stable cell lines were generated with replication-competent chimeras for in vitro antiviral activity determination of HCV nonnucleoside polymerase inhibitors (NNIs) that target different regions of the protein. Compounds that bind to the NNI2 (thiophene carboxylic acid) or NNI3 (benzothiadiazine) allosteric sites showed 8- to >1,280-fold reductions in antiviral activity against non-GT1 NS5B chimeric replicons compared to that against the GT1b subgenomic replicon. Smaller reductions in susceptibility, ranging from 0.2- to 33-fold, were observed for the inhibitor binding to the NNI1 (benzimidazole) site. The inhibitor binding to the NNI4 (benzofuran) site showed broad-spectrum antiviral activity against all chimeric replicons evaluated in this study. In conclusion, evaluation of HCV NNIs against intergenotypic chimeric replicons showed differences in activity spectrum for inhibitors that target different regions of the enzyme, some of which could be associated with specific residues that differ between GT1 and non-GT1 polymerases. Our study demonstrates the utility of chimeric replicons for broad-spectrum activity determination of HCV inhibitors.

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A Small Molecule Inhibits Virion Attachment to Heparan Sulfate- or Sialic Acid-Containing Glycans

Journal of Virology ◽

10.1128/jvi.00896-14 ◽

2014 ◽

Vol 88 (14) ◽

pp. 7806-7817 ◽

Cited By ~ 60

Author(s):

Che C. Colpitts ◽

Luis M. Schang

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Sialic Acid ◽

Herpes Simplex ◽

Heparan Sulfate ◽

Small Molecule ◽

Broad Spectrum ◽

Common Mechanism ◽

Human Viruses ◽

Hsv 1

ABSTRACTPrimary attachment to cellular glycans is a critical entry step for most human viruses. Some viruses, such as herpes simplex virus type 1 (HSV-1) and hepatitis C virus (HCV), bind to heparan sulfate, whereas others, such as influenza A virus (IAV), bind to sialic acid. Receptor mimetics that interfere with these interactions are active against viruses that bind to either heparan sulfate or to sialic acid. However, no molecule that inhibits the attachment of viruses in both groups has yet been identified. Epigallocatechin gallate (EGCG), a green tea catechin, is active against many unrelated viruses, including several that bind to heparan sulfate or to sialic acid. We sought to identify the basis for the broad-spectrum activity of EGCG. Here, we show that EGCG inhibits the infectivity of a diverse group of enveloped and nonenveloped human viruses. EGCG acts directly on the virions, without affecting the fluidity or integrity of the virion envelopes. Instead, EGCG interacts with virion surface proteins to inhibit the attachment of HSV-1, HCV, IAV, vaccinia virus, adenovirus, reovirus, and vesicular stomatitis virus (VSV) virions. We further show that EGCG competes with heparan sulfate for binding of HSV-1 and HCV virions and with sialic acid for binding of IAV virions. Therefore, EGCG inhibits unrelated viruses by a common mechanism. Most importantly, we have identified EGCG as the first broad-spectrum attachment inhibitor. Our results open the possibility for the development of small molecule broad-spectrum antivirals targeting virion attachment.IMPORTANCEThis study shows that it is possible to develop a small molecule antiviral or microbicide active against the two largest groups of human viruses: those that bind to glycosaminoglycans and those that bind to sialoglycans. This group includes the vast majority of human viruses, including herpes simplex viruses, cytomegalovirus, influenza virus, poxvirus, hepatitis C virus, HIV, and many others.

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Dasabuvir inhibits human norovirus infection in human intestinal enteroids

10.1101/2021.07.02.450857 ◽

2021 ◽

Author(s):

Tsuyoshi Hayashi ◽

Kosuke Murakami ◽

Junki Hirano ◽

Yoshiki Fujii ◽

Yoko Yamaoka ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Severe Acute Respiratory Syndrome ◽

Culture System ◽

Age Groups ◽

Viral Gastroenteritis ◽

Compound Library ◽

Human Norovirus ◽

Antiviral Compound ◽

Acute Viral Gastroenteritis

Human noroviruses (HuNoVs) are acute viral gastroenteritis pathogens that affect all age groups, yet no approved vaccines and drugs to treat HuNoV infection are available. In this study, with a human intestinal enteroid (HIE) culture system where HuNoVs are able to replicate reproducibly, we screened an antiviral compound library to identify compound(s) showing anti-HuNoV activity. Dasabuvir, which has been developed as an anti-hepatitis C virus agent, was found to inhibit HuNoV infection in HIEs at micromolar concentrations. Dasabuvir also inhibited severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and human A rotavirus (RVA) infection in HIEs. To our knowledge, this is the first study to screen an antiviral compound library for HuNoV using HIEs and we successfully identified dasabuvir as a novel anti-HuNoV inhibitor that warrants further investigation.

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Imino sugars inhibit the formation and secretion of bovine viral diarrhea virus, a pestivirus model of hepatitis C virus: Implications for the development of broad spectrum anti-hepatitis virus agents

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.96.21.11878 ◽

1999 ◽

Vol 96 (21) ◽

pp. 11878-11882 ◽

Cited By ~ 231

Author(s):

N. Zitzmann ◽

A. S. Mehta ◽

S. Carrouee ◽

T. D. Butters ◽

F. M. Platt ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Broad Spectrum ◽

Bovine Viral Diarrhea Virus ◽

Hepatitis Virus ◽

Bovine Viral Diarrhea ◽

Diarrhea Virus ◽

Viral Diarrhea ◽

Viral Diarrhea Virus

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Identification of a Potent and Broad-Spectrum Hepatitis C Virus Fusion Inhibitory Peptide from the E2 Stem Domain

Scientific Reports ◽

10.1038/srep25224 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 6

Author(s):

Xiaojing Chi ◽

Yuqiang Niu ◽

Min Cheng ◽

Xiuying Liu ◽

Yetong Feng ◽

...

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Broad Spectrum ◽

Inhibitory Peptide ◽

Virus Fusion

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Identification of Hepatitis C Virus Inhibitors Targeting Different Aspects of Infection Using a Cell-Based Assay

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01413-12 ◽

2012 ◽

Vol 56 (12) ◽

pp. 6109-6120 ◽

Cited By ~ 6

Author(s):

Xuemei Yu ◽

Bruno Sainz ◽

Pavel A. Petukhov ◽

Susan L. Uprichard

Keyword(s):

Hepatitis C Virus ◽

Hepatitis C ◽

Viral Infection ◽

Treatment Options ◽

Hcv Infection ◽

Antiviral Compound ◽

Worldwide Population ◽

Dependent Inhibition ◽

Compound Screening ◽

A Cell

ABSTRACTWith 2 to 3% of the worldwide population chronically infected, hepatitis C virus (HCV) infection continues to be a major health care burden. Unfortunately, current interferon-based treatment options are not effective in all patients and are associated with significant side effects. Consequently, there is an ongoing need to identify and develop new anti-HCV therapies. Toward this goal, we previously developed a cell-based HCV infection assay for antiviral compound screening based on a low-multiplicity-of-infection approach that uniquely allows for the identification of antiviral compounds that target cell culture-derived HCV (HCVcc) at any step of the viral infection cycle. Using this assay, here we report the screening of the NCI Diversity Set II library, containing 1,974 synthesized chemical compounds, and the identification of compounds with specific anti-HCV activity. In combination with toxicity counterscreening, we identified 30 hits from the compound library, 13 of which showed reproducible and dose-dependent inhibition of HCV with mean therapeutic indices (50% cytotoxic concentration [CC50]/50% effective concentration [EC50]) of greater than 6. Using HCV pseudotype and replicon systems of multiple HCV genotypes, as well as infectious HCVcc-based assembly and secretion analysis, we determined that different compounds within this group of candidate inhibitors target different steps of viral infection. The compounds identified not only will serve as biological probes to study and further dissect the biology of viral infection but also should facilitate the development of new anti-HCV therapeutic treatments.

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