FDA Approved Drugs Efavirenz, Tipranavir, and Dasabuvir Inhibit Replication of Multiple Flaviviruses In Vitro

Arthropod-borne flaviviruses such as tick-borne encephalitis virus (TBEV), West Nile virus (WNV), Zika virus (ZIKV), Dengue virus (DENV), and yellow fever virus (YFV) cause several serious life-threatening syndromes (encephalitis, miscarriages, paralysis, etc.). No effective antiviral therapy against these viruses has been approved yet. We selected, via in silico modeling, 12 U.S. Food and Drug Administration (FDA)-approved antiviral drugs (paritaprevir, dolutegravir, raltegravir, efavirenz, elvitegravir, tipranavir, saquinavir, dasabuvir, delavirdine, maraviroc, trifluridine, and tauroursodeoxycholic acid) for their interaction with ZIKV proteins (NS3 helicase and protease, non-structural protein 5 (NS5) RNA-dependent RNA polymerase, and methyltransferase). Only three of them were active against ZIKV, namely, dasabuvir (ABT-333), efavirenz, and tipranavir. These compounds inhibit virus replication of ZIKV (MR-766 and Paraiba_01) in Vero cells; therefore, we tested these compounds against other medically important flaviviruses WNV (13-104 and Eg101) and TBEV (Hypr). Dasabuvir was originally developed as an antiviral drug against hepatitis C virus (HCV); tipranavir and efavirenz are used for treating human immunodeficiency virus (HIV) infection. The antiviral effects of efavirenz, tipranavir, and dasabuvir were tested for ZIKV in HUH-7, astrocytes (HBCA), and UKF-NB-4 cells, where we also identified a significant inhibition effect of these compounds. For Vero cells, efavirenz inhibited all investigated viruses with EC50 ranging from 9.70 to 29.26 µM; the tipranavir inhibition effect was from 16.19 (WNV 13-104) to 27.47 µM (TBEV), while the strongest and most robust antiviral effect was demonstrated in the case of dasabuvir (EC50 values ranging from 9.09 (TBEV) to 10.85 µM (WNV 13-104)). These results warrant further research of these drugs, either individually or in combination, as possible pan-flavivirus inhibitors.

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Caspofungin and LTX-315 inhibit SARS-CoV-2 replication by targeting the nsp12 polymerase

10.21203/rs.3.rs-19872/v1 ◽

2020 ◽

Author(s):

Min Wang ◽

Fei Ye ◽

Jiaqi Su ◽

Jingru Zhao ◽

Bin Yuan ◽

...

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Vero Cells ◽

Polymerase Activity ◽

Structural Protein ◽

Live Virus ◽

Drug Candidates ◽

Virtual Screen ◽

Virus Assay ◽

Promising Target

Abstract The ongoing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, previously designated as 2019-nCoV) outbreak has caused global concern1. Currently, there are no clinically approved specific drugs or vaccines available for this virus. The viral polymerase is a promising target for developing broad- spectrum antiviral drugs. Here, based on the highly similar structure of SARS- CoV non-structural protein 12 (nsp12) polymerase subunit2, we applied virtual screen for the available compounds, including both the FDA-approved and under- clinic drugs, to identify potential antiviral molecules against SARS-CoV-2. We found two drugs, the clinically approved anti-fungi drug Caspofungin Acetate (Cancidas) and the oncolytic peptide LTX-315, can bind SARS-CoV-2 nsp12 protein to block the polymerase activity in vitro. Further live virus assay revealed that both Caspofungin Acetate and LTX-315 can effectively inhibit SARS-CoV-2 replication in vero cells. These findings present promising drug candidates for treatment of related diseases and would also stimulate the development of pan- coronavirus antiviral agents.Authors Min Wang, Fei Ye, Jiaqi Su, Jingru Zhao, and Bin Yuan contributed equally to this work.

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Investigation of the effects of six medicinal plants with antiviral effects against COVID-19

10.21203/rs.3.rs-766824/v1 ◽

2021 ◽

Author(s):

Harun ALP ◽

Hasan ASİL ◽

Demet Duman

Keyword(s):

Medicinal Plants ◽

Nigella Sativa ◽

Water Extract ◽

Antiviral Effect ◽

Vero Cells ◽

Glycyrrhiza Glabra ◽

Crocus Sativus ◽

Medicinal And Aromatic Plants ◽

The World

Abstract Today, the coronavirus epidemic, which caused the death of 79 million cases and 1743 thousand people in 218 countries around the world, continues to increase its impact all over the world. Researchers are still trying to develop an effective solution against covid-19, including vaccines and drugs. However, there are few studies that determine the effect of natural products obtained from plants on covid-19. Medicinal and aromatic plants have been used for therapeutic purposes since the existence of humanity. In this study, the effects of some important medicinal plants including Licorice (Glycyrrhiza glabra), Saffron (Crocus sativus L.), Nigella (Nigella sativa L.), Laurel (Lauris nobilis), Karabaş (Lavandula stoechas), and Zahter (Thymbra spicata L. var. Spicata) against Covid-19 were investigated in vitro conditions. The six plants were evaluated for cytotoxic effect on Vero cells and determining inhibition of viral replication in Vero-E6 cells at concentrations of broad-spectrum antiviral non-cytotoxic against Covid-19 in cell culture and an additional antiviral effect against Covid-19. According to the results, the five examined plants (Saffron, Nigella, Laurel, Karabaş, Zahter) were ineffective against Covid-19 in vitro conditions. Interisingly, the water extract obtained from the root of the licorice plant (Glycyrrhiza glabra) inhibited Covid-19 in vitro conditions in the 2nd dilution (1: 4) following the initial concentration in Vero-E6 cells.

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Infection of SCID mice with Montana Myotis leukoencephalitis virus as a model for flavivirus encephalitis

Journal of General Virology ◽

10.1099/0022-1317-83-8-1887 ◽

2002 ◽

Vol 83 (8) ◽

pp. 1887-1896 ◽

Cited By ~ 19

Author(s):

Nathalie Charlier ◽

Pieter Leyssen ◽

Jan Paeshuyse ◽

Christian Drosten ◽

Herbert Schmitz ◽

...

Keyword(s):

Yellow Fever Virus ◽

Antiviral Agents ◽

Antiviral Drug ◽

Vero Cells ◽

Scid Mice ◽

Fever Virus ◽

Viral Rna ◽

Interferon Α ◽

Immunocompetent Mice ◽

Antiviral Targets

We have established a convenient animal model for flavivirus encephalitis using Montana Myotis leukoencephalitis virus (MMLV), a bat flavivirus. This virus has the same genomic organization, and contains the same conserved motifs in genes that encode potential antiviral targets, as flaviviruses that cause disease in man (N. Charlier et al., accompanying paper), and has a similar particle size (approximately 40 nm). MMLV replicates well in Vero cells and appears to be equally as sensitive as yellow fever virus and dengue fever virus to a selection of experimental antiviral agents. Cells infected with MMLV show dilation of the endoplasmic reticulum, a characteristic of flavivirus infection. Intraperitoneal, intranasal or direct intracerebral inoculation of SCID mice with MMLV resulted in encephalitis ultimately leading to death, whereas immunocompetent mice were refractory to either intranasal or intraperitoneal infection with MMLV. Viral RNA and/or antigens were detected in the brain and serum of MMLV-infected SCID mice, but not in any other organ examined: MMLV was detected in the olfactory lobes, the cerebral cortex, the limbic structures, the midbrain, cerebellum and medulla oblongata. Infection was confined to neurons. Treatment with the interferon-α/β inducer poly(I)·poly(C) protected SCID mice against MMLV-induced morbidity and mortality, and this protection correlated with a reduction in infectious virus titre and viral RNA load. This validates the MMLV model for use in antiviral drug studies. The MMLV SCID model may, therefore, be attractive for the study of chemoprophylactic or chemotherapeutic strategies against flavivirus infections causing encephalitis.

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Decoy Receptor Interactions as Novel Drug Targets against EKC-Causing Human Adenovirus

Viruses ◽

10.3390/v11030242 ◽

2019 ◽

Vol 11 (3) ◽

pp. 242 ◽

Cited By ~ 5

Author(s):

Naresh Chandra ◽

Lars Frängsmyr ◽

Niklas Arnberg

Keyword(s):

Drug Targets ◽

Antiviral Drug ◽

Human Adenovirus ◽

Adenovirus Type ◽

Antiviral Effect ◽

Epidemic Keratoconjunctivitis ◽

Virus Binding ◽

Decoy Receptor ◽

Decoy Receptors ◽

Approved Drugs

Epidemic keratoconjunctivitis (EKC) is a severe ocular disease and can lead to visual impairment. Human adenovirus type-37 (HAdV-D37) is one of the major causative agents of EKC and uses sialic acid (SA)-containing glycans as cellular receptors. Currently, there are no approved antivirals available for the treatment of EKC. Recently, we have reported that sulfated glycosaminoglycans (GAGs) bind to HAdV-D37 via the fiber knob (FK) domain of the viral fiber protein and function as decoy receptors. Based on this finding, we speculated that GAG-mimetics may act as artificial decoy receptors and inhibit HAdV-D37 infection. Repurposing of approved drugs to identify new antivirals has drawn great attention in recent years. Here, we report the antiviral effect of suramin, a WHO-approved drug and a widely known GAG-mimetic, against HAdV-D37. Commercially available suramin analogs also show antiviral effects against HAdV-D37. We demonstrate that suramin exerts its antiviral activity by inhibiting the attachment of HAdV-D37 to cells. We also reveal that the antiviral effect of suramin is HAdV species-specific. Collectively, in this proof of concept study, we demonstrate for the first time that virus binding to a decoy receptor constitutes a novel and an unexplored target for antiviral drug development.

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Adaptation of yellow fever virus 17D to Vero cells is associated with mutations in structural and non-structural protein genes

Virus Research ◽

10.1016/j.virusres.2013.04.003 ◽

2013 ◽

Vol 176 (1-2) ◽

pp. 280-284 ◽

Cited By ~ 8

Author(s):

David W.C. Beasley ◽

Merribeth Morin ◽

Ashley R. Lamb ◽

Edward Hayman ◽

Douglas M. Watts ◽

...

Keyword(s):

Yellow Fever ◽

Yellow Fever Virus ◽

Vero Cells ◽

Fever Virus ◽

Structural Protein

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IFN-λ1 Displays Various Levels of Antiviral Activity In Vitro in a Select Panel of RNA Viruses

Viruses ◽

10.3390/v13081602 ◽

2021 ◽

Vol 13 (8) ◽

pp. 1602

Author(s):

Marina Plotnikova ◽

Alexey Lozhkov ◽

Ekaterina Romanovskaya-Romanko ◽

Irina Baranovskaya ◽

Mariia Sergeeva ◽

...

Keyword(s):

Antiviral Activity ◽

Rna Viruses ◽

Antiviral Effect ◽

Vero Cells ◽

Genome Replication ◽

Type I ◽

Protective Effects ◽

Cellular Models ◽

Viral Genome Replication

Type III interferons (lambda IFNs) are a quite new, small family of three closely related cytokines with interferon-like activity. Attention to IFN-λ is mainly focused on direct antiviral activity in which, as with IFN-α, viral genome replication is inhibited without the participation of immune system cells. The heterodimeric receptor for lambda interferons is exposed mainly on epithelial cells, which limits its possible action on other cells, thus reducing the likelihood of developing undesirable side effects compared to type I IFN. In this study, we examined the antiviral potential of exogenous human IFN-λ1 in cellular models of viral infection. To study the protective effects of IFN-λ1, three administration schemes were used: ‘preventive’ (pretreatment); ‘preventive/therapeutic’ (pre/post); and ‘therapeutic’ (post). Three IFN-λ1 concentrations (from 10 to 500 ng/mL) were used. We have shown that human IFN-λ1 restricts SARS-CoV-2 replication in Vero cells with all three treatment schemes. In addition, we have shown a decrease in the viral loads of CHIKV and IVA with the ‘preventive’ and ‘preventive/therapeutic’ regimes. No significant antiviral effect of IFN-λ1 against AdV was detected. Our study highlights the potential for using IFN-λ as a broad-spectrum therapeutic agent against respiratory RNA viruses.

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Arbidol (Umifenovir): A Broad-spectrum Antiviral Drug that Inhibits Medically Important Arthropod-borne Flaviviruses

10.20944/preprints201802.0134.v1 ◽

2018 ◽

Author(s):

Jan Haviernik ◽

Michal Stefanik ◽

Martina Fojtikova ◽

Sabrina Kali ◽

Noël Tordo ◽

...

Keyword(s):

Broad Spectrum ◽

Antiviral Drug ◽

Neuroblastoma Cells ◽

Antiviral Effect ◽

Vero Cells ◽

Cell Interaction ◽

Human Pathogens ◽

Human Hepatoma Cells ◽

Human Neuroblastoma ◽

Antiviral Compound

Arthropod-borne flaviviruses represent human pathogens of global medical importance, against which no effective small molecule-based antiviral therapy is currently available. Arbidol (umifenovir) is a broad spectrum antiviral compound approved in Russia and China for prophylaxis and treatment of influenza. This compound showed activity against numerous DNA and RNA viruses. Its mode of action is based predominantly on the impairment of critical steps of virus-cell interaction. Here we demonstrate that arbidol possesses a micromolar inhibition activity (EC50 values ranging from 10.57 ± 0.74 to 19.16 ± 0.29 µM) in Vero cells infected with Zika virus, West Nile virus, and tick-borne encephalitis virus, three medically important representatives of arthropod-borne flaviviruses. Interestingly, no antiviral effect of arbidol is observed in porcine stable kidney cells (PS), human neuroblastoma cells (UKF-NB-6), human hepatoma cells (Huh-7 cells) indicating that the antiviral effect of arbidol is strongly cell-type dependent. Arbidol presents a significant increasing in cytotoxicity profiles when tested in various cell lines in the order: Huh-7 < HBCA < PS < UKF-NB-6 < Vero with CC50 values ranging from 18.69 ± 0.1 to 89.72 ± 0.19 µM. Antiviral activity and acceptable cytotoxicity profiles suggest that arbidol could be a promising candidate for further investigation as a potential therapeutic agent in treating flaviviral infections.

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In vitro Assessment of Antiviral Effect of Natural Compounds on Porcine Epidemic Diarrhea Coronavirus

Frontiers in Veterinary Science ◽

10.3389/fvets.2021.652000 ◽

2021 ◽

Vol 8 ◽

Author(s):

Manuel Gómez-García ◽

Héctor Puente ◽

Héctor Argüello ◽

Óscar Mencía-Ares ◽

Pedro Rubio ◽

...

Keyword(s):

Antiviral Activity ◽

Formic Acid ◽

Antiviral Agents ◽

Antiviral Effect ◽

Vero Cells ◽

Optical Microscope ◽

Dependent Manner ◽

Diarrhea Virus ◽

Porcine Epidemic Diarrhea

Organic acid and essential oils (EOs), well-known antimicrobials, could also possess antiviral activity, a characteristic which has not been completely addressed up to now. In this study, the effect of two organic acids (formic acid and sodium salt of coconut fatty acid distillates) and two single EO compounds (thymol and cinnamaldehye) was evaluated against porcine epidemic diarrhea virus (PEDV). The concentration used for each compound was established by cytotoxicity assays in Vero cells. The antiviral activity was then evaluated at three multiplicities of infection (MOIs) through visual cytopathic effect (CPE) evaluation and an alamarBlue assay as well as real-time reverse-transcription PCR (RT-qPCR) and viral titration of cell supernatants. Formic acid at at a dose of 1,200 ppm was the only compound which showed antiviral activity, with a weak reduction of CPE caused by PEDV. Through the alamarBlue fluorescence assay, we showed a significant anti-CPE effect of formic acid which could not be observed by using an inverted optical microscope. RT-qPCR and infectivity analysis also showed that formic acid significantly reduced viral RNA and viral titers in a PEDV MOI-dependent manner. Our results suggest that the antiviral activity of formic acid could be associated to its inhibitory effect on viral replication. Further studies are required to explore the anti-PEDV activity of formic acid under field conditions alone or together with other antiviral agents.

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Inhibition of Arenaviruses by Combinations of Orally Available Approved Drugs

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01146-20 ◽

2021 ◽

Author(s):

Shawn Herring ◽

Jessica M. Oda ◽

Jessica Wagoner ◽

Delaney Kirchmeier ◽

Aidan O’Connor ◽

...

Keyword(s):

Virus Entry ◽

Antiviral Drug ◽

Lymphocytic Choriomeningitis ◽

Neglected Diseases ◽

Proof Of Concept ◽

Effective Suppression ◽

Resource Limited ◽

Approved Drugs ◽

And Control

Neglected diseases caused by arenaviruses such as Lassa (LASV) and filoviruses like Ebola (EBOV) primarily afflict resource-limited countries, where antiviral drug development is often minimal. Previous studies have shown that many approved drugs developed for other clinical indications inhibit EBOV and LASV and that combinations of these drugs provide synergistic suppression of EBOV, often by blocking discrete steps in virus entry. We hypothesize that repurposing combinations of orally administered approved drugs provides effective suppression of arenaviruses. In this report, we demonstrate that arbidol, an approved influenza antiviral previously shown to inhibit EBOV, LASV, and many other viruses, inhibits murine leukemia (MLV) reporter viruses pseudotyped with the fusion glycoproteins (GP) of other arenaviruses [Junin (JUNV), lymphocytic choriomeningitis virus (LCMV), and Pichinde (PICV)]. Arbidol and other approved drugs including aripiprazole, amodiaquine, sertraline, and niclosamide also inhibit infection of cells by infectious PICV, and arbidol, sertraline and niclosamide inhibit infectious LASV. Combining arbidol with aripiprazole or sertraline results in synergistic suppression of LASV and JUNV GP-bearing pseudoviruses. This proof-of-concept study shows that arenavirus infection in vitro can be synergistically inhibited by combinations of approved drugs. This approach may lead to a proactive strategy with which to prepare for and control known and new arenavirus outbreaks.

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Déjà vu: Stimulating Open Drug Discovery for SARS-CoV-2

10.31219/osf.io/hczra ◽

2020 ◽

Author(s):

Sean Ekins ◽

Melina Mottin ◽

Paulo R. P. S. Ramos ◽

Bruna K. P. Sousa ◽

Bruno Junior Neves ◽

...

Keyword(s):

Drug Discovery ◽

Ebola Virus ◽

Antiviral Drug ◽

Open Science ◽

Virus Rna ◽

Deja Vu ◽

Déjà Vu ◽

Approved Drugs

In the past decade we have seen two major Ebola virus outbreaks in Africa, the Zika virus in Brazil and the current outbreak of coronavirus disease which has been named "severe acute respiratory syndrome coronavirus 2" (SARS-CoV-2). There is a strong sense of Déjà vu as the world is caught flat footed without effective treatments to administer to patients. Our team has been actively involved in several small molecule drug discovery efforts for the preceding virus outbreaks. In 2014 we used machine learning to identify 3 new molecules to test for the Ebola virus and these were subsequently shown to be active in vitro and in vivo. We have also been involved in open science approaches that leverage the community to help. In 2016 we launched the OpenZika project as an IBM World Community Grid Project that used distributed computing power of volunteers to dock large numbers of compounds into Zika and related flavivirus targets. This led us into several collaborations in which we validated computational predictions in vitro. With both of these initiatives there was some knowledge of the virus, many compounds had already been tested in the case of Ebola, whereas for Zika initially all we had was the virus RNA sequence. In the current SARS-CoV-2 outbreak, this was a completely new virus and the scientists in China and elsewhere have started from scratch. In the space of a few weeks since the outbreak is acknowledged to have started, there are now compounds suggested as active in vitro and molecules repurposed in clinical trials. While this has been impressive, we propose there may still be gaps in our approach to drug discovery for such outbreaks. There is an opportunity to repurpose additional approved drugs for this virus and we now suggest how these might be identified leveraging prior work on MERS-CoV, SARS-CoV and other viruses. We also describe some of the immense challenges and limitations of the open antiviral drug discovery approaches we have been involved in.

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