scholarly journals The clinically approved antiviral drug sofosbuvir impairs brazilian zika virus replication

2016 ◽  
Author(s):  
Caroline Q. Sacramento ◽  
Gabrielle R. de Melo ◽  
Natasha Rocha ◽  
Lucas Villas Bôas Hoelz ◽  
Milene Mesquita ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Zika Virus ◽  
Antiviral Drug ◽  
Time Frame ◽  
Polymerase Activity ◽  
Antiviral Drugs ◽  
South American ◽  
Dependent Manner ◽  
Amino Acid Residues ◽  
Uridine Nucleotide

SummaryZika virus (ZIKV) is a member of Flaviviridae family, as other agents of clinical significance, such as dengue (DENV) and hepatitis C (HCV) viruses. ZIKV spread from Africa to Pacific and South American territories, emerging as an etiological pathogen of neurological disorders, during fetal development and in adulthood. Therefore, antiviral drugs able to inhibit ZIKV replication are necessary. Broad spectrum antivirals, such as interferon, ribavirin and favipiravir, are harmful for pregnant animal models and women. The clinically approved uridine nucleotide analog anti-HCV drug, sofosbuvir, has not been affiliated to teratogenicity. Sofosbuvir target the most conserved protein over the members of the Flaviviridae family, the viral RNA polymerase. We thus studied ZIKV susceptibility to sofosbovir. We initially characterized a Brazilian ZIKV strain for use in experimental assays. Sofosbuvir inhibits the Brazilian ZIKV replication in a dose-dependent manner, both in BHK-21 cells and SH-Sy5y, by targeting ZIKV RNA polymerase activity, with the involvement of conserved amino acid residues over the members of Flaviviridae family. The identification of clinically approved antiviral drugs endowed with anti-ZIKV could reduce the time frame in pre-clinical development. Altogether, our data indicates that sofosbuvir chemical structure is endowed with anti-ZIKV activity.

scholarly journals Non-nucleoside Inhibitors of Zika Virus RNA-Dependent RNA Polymerase

2020 ◽  
Vol 94 (21) ◽  
Author(s):  
Aicha Gharbi-Ayachi ◽  
Sridhar Santhanakrishnan ◽  
Yee Hwa Wong ◽  
Kitti W. K. Chan ◽  
Siok Thing Tan ◽  
...  
Keyword(s):  
High Resolution ◽  
Dengue Virus ◽  
Rna Polymerase ◽  
Zika Virus ◽  
Nonstructural Protein ◽  
Specific Binding ◽  
Antiviral Drug ◽  
Polymerase Activity ◽  
Health Concern ◽  
Rna Dependent Rna Polymerase

ABSTRACT Zika virus (ZIKV) remains a potentially significant public health concern because it can cause teratogenic effects, such as microcephaly in newborns and neurological disease, like Guillain-Barré syndrome. Together with efforts to develop a vaccine, the discovery of antiviral molecules is important to control ZIKV infections and to prevent its most severe symptoms. Here, we report the development of small nonnucleoside inhibitors (NNIs) of ZIKV RNA-dependent RNA polymerase (RdRp) activity. These NNIs target an allosteric pocket (N pocket) located next to a putative hinge region between the thumb and the palm subdomains that was originally described for dengue virus (DENV) RdRp. We first tested the activity of DENV RdRp N-pocket inhibitors against ZIKV RdRp, introduced chemical modifications into these molecules, and assessed their potency using both enzymatic and cell-based assays. The most potent compound had a 50% inhibitory concentration value of 7.3 μM and inhibited ZIKV replication in a cell-based assay with a 50% effective concentration value of 24.3 μM. Importantly, we report four high-resolution crystal structures detailing how these NNIs insert into the N pocket of ZIKV RdRp. Our observations point to subtle differences in the size, shape, chemical environment, and hydration of the N pocket from ZIKV RdRp from those of the N pocket from DENV RdRp that are crucial for the design of improved antiviral inhibitors with activity against ZIKV. IMPORTANCE Zika virus belongs to the Flavivirus genus, which comprises several important human pathogens. There is currently neither an approved vaccine nor antiviral drugs available to prevent infection by ZIKV. The nonstructural protein 5 (NS5) polymerase, which is responsible for replicating the viral RNA genome, represents one of the most promising targets for antiviral drug development. Starting from compounds recently developed against dengue virus NS5, we designed and synthesized inhibitors targeting Zika virus NS5. We show that these novel compounds inhibit viral replication by targeting the polymerase activity. High-resolution X-ray crystallographic structures of protein-inhibitor complexes demonstrated specific binding to an allosteric site within the polymerase, called the N pocket. This work paves the way for the future structure-based design of potent compounds specifically targeting ZIKV RNA polymerase activity.

scholarly journals The clinically approved antiviral drug sofosbuvir inhibits Zika virus replication

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Carolina Q. Sacramento ◽  
Gabrielle R. de Melo ◽  
Caroline S. de Freitas ◽  
Natasha Rocha ◽  
Lucas Villas Bôas Hoelz ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Zika Virus ◽  
Neurological Disorders ◽  
Viral Genome ◽  
Antiviral Drug ◽  
Cellular Systems ◽  
Viral Rna ◽  
Amino Acid Residues ◽  
Direct Inhibition ◽  
Secondary Use

Abstract Zika virus (ZIKV) is a member of the Flaviviridae family, along with other agents of clinical significance such as dengue (DENV) and hepatitis C (HCV) viruses. Since ZIKV causes neurological disorders during fetal development and in adulthood, antiviral drugs are necessary. Sofosbuvir is clinically approved for use against HCV and targets the protein that is most conserved among the members of the Flaviviridae family, the viral RNA polymerase. Indeed, we found that sofosbuvir inhibits ZIKV RNA polymerase, targeting conserved amino acid residues. Sofosbuvir inhibited ZIKV replication in different cellular systems, such as hepatoma (Huh-7) cells, neuroblastoma (SH-Sy5y) cells, neural stem cells (NSC) and brain organoids. In addition to the direct inhibition of the viral RNA polymerase, we observed that sofosbuvir also induced an increase in A-to-G mutations in the viral genome. Together, our data highlight a potential secondary use of sofosbuvir, an anti-HCV drug, against ZIKV.

scholarly journals Beyond members of the Flaviviridae family, sofosbuvir also inhibits chikungunya virus replication

2018 ◽  
Author(s):  
André C. Ferreira ◽  
Patrícia A. Reis ◽  
Caroline S. de Freitas ◽  
Carolina Q. Sacramento ◽  
Lucas Villas Bôas Hoelz ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Chikungunya Virus ◽  
Therapeutic Option ◽  
Antiviral Drug ◽  
Specific Treatment ◽  
Public Health Problem ◽  
Hepatoma Cells ◽  
Dependent Manner ◽  
Mice Model ◽  
Human Hepatoma Cells

AbstractChikungunya virus (CHIKV) causes a febrile disease associated with chronic arthralgia, which may progress to neurological impairment. Chikungunya fever (CF) is a consolidated public health problem, in tropical and subtropical regions of the world, where control of CHIKV vector, mosquitos of theAedesgenus, failed. Since there is no vaccine or specific treatment against CHIKV, infected patients receive only palliative care to alleviate pain and arthralgia. Thus, drug repurposing is necessary to identify antivirals against CHIKV. Recently, the structure and activity of CHIKV RNA polymerase was partially resolved, revealing similar aspects with the enzyme counterparner on other positive sense RNA viruses, such as members of the Flaviviridae family. We then evaluated if sofosbuvir, clinically approved against hepatitis C virus RNA polymerase, which also aims to dengue, Zika and yellow fever viruses replication, would inhibit CHIKV replication. Indeed, sofosbuvir was 5-times more selective in inhibiting CHIKV production in human hepatoma cells than ribavirin, a pan-antiviral drug. Although CHIKV replication in human induced pluripotent stem cell (iPS)-derived astrocytes was less sensitive to sofosbuvir’s, compared to hepatoma cells – this drug still impaired virus production and cell death in a MOI-dependent manner. Sofosbuvir also exhibited antiviral activityin vivo, by preventing CHIKV-induced paw oedeme in adult mice, at 20 mg/kg/day, and mortality on neonate mice model, at 40 and 80 mg/kg/day. Our data demonstrates that a prototypic alphavirus, CHIKV, is also susceptible to sofosbuvir. Since this is a clinically approved drug, it could pave the way to become a therapeutic option against CF.

scholarly journals Beclabuvir can Inhibit the RNA-dependent RNA Polymerase of Newly Emerged Novel Coronavirus (SARS-CoV-2)

2020 ◽  
Author(s):  
Kunal Dutta ◽  
Sergey Shityakov ◽  
Olga Morozova ◽  
Ibrahim Khalifa ◽  
Jin Zhang ◽  
...  
Keyword(s):  
Amino Acid ◽  
Rna Polymerase ◽  
Rna Synthesis ◽  
Binding Free Energy ◽  
Polymerase Activity ◽  
Amino Acid Residues ◽  
Rna Dependent Rna Polymerase ◽  
Recent Emergence ◽  
Novel Coronavirus ◽  
Major Amino Acid

Recent emergence of novel coronavirus (SARS-CoV-2) all over the world has resulted more than 33,106 global deaths. To date well-established therapeutics modules for infected patients are unknown. In this present initiative, molecular interactions between FDA-approved antiviral drugs against the Hepatitis-C virus (HCV) have been investigated theoretically against the RNA-dependent RNA polymerase (RdRp) of SARS-CoV-2. HCV and SARS-CoV-2 are both +ssRNA viruses. At 25o C beclabuvir, a non-nucleoside inhibitor of the RdRpHCV can efficiently bind to RdRp SARS-CoV-2 (ΔGAutoDock = -9.95 kcal mol-1) with an inhibition constant of 51.03 nM. Both the ΔGLondon and ΔGGBVI / WSA values were - 9.06 and - 6.67 kcal mol-1, respectively for binding of beclabuvir to RdRpSARS-CoV-2. In addition, beclabuvir has also shown better binding free energy with RdRpSARS-CoV-2 (ΔGvina = -8.0 kcal mol-1) than that observed with the Thumb 1 domain of RdRpHCV (ΔGvina = -7.1 kcal mol-1). InterProScan has suggested the RNA-directed 5'-3' polymerase activity exists within 549th to 776th amino acid residues of RdRpSARS-CoV, where the major amino acid residues interacting being I591, Y621, C624, D625, A690, N693, L760, D762, D763, and E813-N817. Molecular interaction suggests occupancy of beclabuvir inside the active site environment of the RdRpSARS-CoV-2, the enzyme essential for viral RNA synthesis. In conclusion, results suggest beclabuvir may serve as an anti-SARS-CoV-2 drug.

scholarly journals Beclabuvir can Inhibit the RNA-dependent RNA Polymerase of Newly Emerged Novel Coronavirus (SARS-CoV-2)

2020 ◽  
Author(s):  
Kunal Dutta ◽  
Sergey Shityakov ◽  
Olga Morozova ◽  
Ibrahim Khalifa ◽  
Jin Zhang ◽  
...  
Keyword(s):  
Amino Acid ◽  
Rna Polymerase ◽  
Rna Synthesis ◽  
Therapeutic Potential ◽  
Binding Free Energy ◽  
Polymerase Activity ◽  
Amino Acid Residues ◽  
Rna Dependent Rna Polymerase ◽  
Recent Emergence ◽  
Novel Coronavirus

Recent emergence of novel coronavirus (SARS-CoV-2) in Wuhan, China has resulted more than 14,510 global deaths. To date well-established therapeutics modules for infected patients are unknown. In this present initiative, molecular interactions between well-known antiviral drugs against the Hepatitis-C virus (HCV) have been investigated theoretically against the RNA-dependent RNA polymerase (RdRp) of SARS-CoV-2. HCV and SARS-CoV-2 are both +ssRNA viruses. At 25o C beclabuvir, a non-nucleoside inhibitor of the RdRp of the HCV can efficiently bind to RdRp of the SARS-CoV-2 (ΔGAutoDock = -9.95 kcal mol-1) with an inhibition constant of only 51.03 nM. Both the ΔGLondon and ΔGGBVI / WSA values were - 9.06 and - 6.67 kcal mol-1, respectively for SARS-CoV-2. In addition, beclabuvir also shows better binding free energy (ΔGvina = 9.7 kcal mol-1) than that of the Thumb 1 domain of RdRp of HCV (ΔGvina = 7.7 kcal mol-1). InterProScan has suggested the RNA-directed 5'-3' polymerase activity existed within 549 to 776 amino acid residues of RdRp. Moreover, major interacting amino acid residues were I591, Y621, C624, D625, A690, N693, L760, D762, D763 and E813-N817. Molecular interaction suggests occupancy of beclabuvir inside the active site environment of the RdRp which is essential for viral RNA synthesis. In conclusion, results suggest beclabuvir has high therapeutic potential as an anti-SARS-CoV-2 drug.

scholarly journals Flavonoids and Nucleotide Analogs Show High Affinity for Viral Proteins of SARS-CoV-2 by in silico Analysis: New Candidates for the Treatment of COVID-19.

2020 ◽  
Author(s):  
Luis Adrián De Jesús-González ◽  
Juan Fidel Osuna-Ramos ◽  
José Manuel Reyes-Ruiz ◽  
Carlos Noe Farfan-Morales ◽  
Selvin Noé Palacios-Rápalo ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Inhibitory Activity ◽  
In Silico ◽  
Antiviral Drug ◽  
World Health ◽  
Nucleotide Analogs ◽  
High Affinity ◽  
Uridine Nucleotide

Abstract The recent epidemic of COVID-19 caused by SARS-CoV-2 was declared by the World Health Organization as a public health emergency of international concern. The absence of an approved vaccine or a specific antiviral drug has made bioinformatic tools crucial for the identification of potential therapeutic targets and drugs for its control. As in other RNA viruses, the protease 3C-like and the RNA-polymerase are two of the SARS-CoV-2 targets to test drugs that can be analyzed in silico. In the present study, compounds derived from plants, fungi, and nucleoside 5'-triphosphate or uridine nucleotide analogs, with anti-DENV activity in vitro or in vivo, were analyzed by molecular docking as potential anti-SARS-CoV-2 drugs. Anthraquinone, with a DENV NS3 protease inhibitory activity; Balapiravir, Fisetin, Hyperoside, and Sofosbuvir, with a DENV NS5 RNA-polymerase inhibitory activity; and Quercetin, with both anti-NS3-NS5 activities, were tested against 3C-like protease and RNA-polymerase of SARS-CoV-2. All these drugs demonstrated a high affinity for the corresponding SARS-CoV-2 proteins, representing excellent candidates for the treatment of COVID-19. Therefore, in vitro or in vivo studies should be carried out using these compounds on models for SARS-CoV-2 infection.

scholarly journals Construction of a non-infectious SARS-CoV-2 replicon for antiviral drug testing and gene function studies

2021 ◽  
Author(s):  
Hai Trong Nguyen ◽  
Darryl Falzarano ◽  
Volker Gerdts ◽  
Qiang Liu
Keyword(s):  
Drug Testing ◽  
Reverse Genetics ◽  
Antiviral Drug ◽  
Antiviral Drugs ◽  
Dependent Manner ◽  
Global Public Health ◽  
Structural Protein ◽  
Reporter System ◽  
Viral Pathogen ◽  
Public Health Emergencies

The emerging coronavirus disease 2019 (COVID-19) outbreak caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has rapidly spread worldwide resulting in global public health emergencies and economic crises. In this study, a non-infectious and biocontainment level-2 compatible SARS-CoV-2 replicon expressing a nano luciferase (nLuc) reporter was constructed in a bacterial artificial chromosomal (BAC) vector by reverse genetics. The nLuc reporter is highly sensitive, easily quantifiable, and high-throughput adaptable. Upon transfecting the SARS-CoV-2 replicon BAC plasmid DNA into Vero E6 cells, we could detect high levels of nLuc reporter activity and viral RNA transcript, suggesting the replication of the replicon. The replicon replication was further demonstrated by the findings that deleting non-structural protein 15 or mutating its catalytic sites significantly reduced replicon replication, whereas providing the nucleocapsid protein in trans enhanced replicon replication in a dose-dependent manner. Finally, we showed that remdesivir, an FDA-approved antiviral drug, significantly inhibited the replication of the replicon, providing proof-of-principle for the application of our replicon as a useful tool for developing antivirals. Taken together, this study established a sensitive and BSL-2 compatible reporter system in a single BAC plasmid for investigating the functions of SARS-CoV-2 proteins in viral replication and evaluating antiviral compounds. This should contribute to the global effort to combat this deadly viral pathogen. IMPORTANCE The COVID-19 pandemic caused by SARS-CoV-2 is having a catastrophic impact on human lives. Combatting the pandemic requires effective vaccines and antiviral drugs. In this study, we developed a SARS-CoV-2 replicon system with a sensitive and easily quantifiable reporter. Unlike studies involving infectious SARS-CoV-2 virus that must be performed in a biosafety level (BSL) 3 facility, the replicon is non-infectious and thus can be safely used in BSL2 laboratories. The replicon will provide a valuable tool for testing antiviral drugs and studying SARS-CoV-2 biology.

scholarly journals Beyond Members of theFlaviviridaeFamily, Sofosbuvir Also Inhibits Chikungunya Virus Replication

2018 ◽  
Vol 63 (2) ◽  
pp. e01389-18 ◽  
Author(s):  
André C. Ferreira ◽  
Patrícia A. Reis ◽  
Caroline S. de Freitas ◽  
Carolina Q. Sacramento ◽  
Lucas Villas Bôas Hoelz ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Virus Replication ◽  
Chikungunya Virus ◽  
Yellow Fever Virus ◽  
Therapeutic Option ◽  
Antiviral Drug ◽  
Specific Treatment ◽  
Public Health Problem ◽  
Hepatoma Cells ◽  
Dependent Manner

ABSTRACTChikungunya virus (CHIKV) causes a febrile disease associated with chronic arthralgia, which may progress to neurological impairment. Chikungunya fever (CF) is an ongoing public health problem in tropical and subtropical regions of the world, where control of the CHIKV vector,Aedesmosquitos, has failed. As there is no vaccine or specific treatment for CHIKV, patients receive only palliative care to alleviate pain and arthralgia. Thus, drug repurposing is necessary to identify antivirals against CHIKV. CHIKV RNA polymerase is similar to the orthologue enzyme of other positive-sense RNA viruses, such as members of theFlaviviridaefamily. Among theFlaviviridae, not only is hepatitis C virus RNA polymerase susceptible to sofosbuvir, a clinically approved nucleotide analogue, but so is dengue, Zika, and yellow fever virus replication. Here, we found that sofosbuvir was three times more selective in inhibiting CHIKV production in human hepatoma cells than ribavirin, a pan-antiviral drug. Although CHIKV replication in human induced pluripotent stem cell-derived astrocytes was less susceptible to sofosbuvir than were hepatoma cells, sofosbuvir nevertheless impaired virus production and cell death in a multiplicity of infection-dependent manner. Sofosbuvir also exhibited antiviral activityin vivoby preventing CHIKV-induced paw edema in adult mice at a dose of 20 mg/kg of body weight/day and prevented mortality in a neonate mouse model at 40- and 80-mg/kg/day doses. Our data demonstrate that a prototypic alphavirus, CHIKV, is also susceptible to sofosbuvir. As sofosbuvir is a clinically approved drug, our findings could pave the way to it becoming a therapeutic option against CF.

scholarly journals SARS-CoV-2 RdRp is a versatile enzyme with proofreading activity and ability to incorporate NHC into RNA by using diphosphate form molnupiravir as a substrate

2021 ◽  
Author(s):  
Maofeng Wang ◽  
Cancan Wu ◽  
Nan Liu ◽  
Fengyu Zhang ◽  
Hongjie Dong ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Drug Development ◽  
Rna Polymerase ◽  
Human Health ◽  
Antiviral Drug ◽  
Polymerase Activity ◽  
Genome Replication ◽  
Rna Dependent Rna Polymerase ◽  
The World ◽  
Rna Polymerase Activity

The coronavirus disease 2019 (COVID-19) has been ravaging throughout the world for almost two years and has severely impaired both human health and the economy. The causative agent, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) employs the viral RNA-dependent RNA polymerase (RdRp) complex for genome replication and transcription, making RdRp an appealing target for antiviral drug development. Although the structure of the RdRp complex has been determined, the function of RdRp has not been fully characterized. Here we reveal that in addition to RNA dependent RNA polymerase activity, RdRp also shows exoribonuclease activity and consequently proofreading activity. We observed that RdRp and nsp14-ExoN, when combined, exhibit higher proofreading activity compared to RdRp alone. Moreover, RdRp can recognize and utilize nucleoside diphosphate (NDP) as substrate to synthesize RNA and can also incorporate β-d-N4-hydroxycytidine (NHC) into RNA while using diphosphate form molnupiravir as substrate.

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