Investigation of monotherapy and combined anticoronaviral therapies against feline coronavirus serotype II in vitro

Objectives Feline infectious peritonitis (FIP), caused by genetic mutants of feline enteric coronavirus known as FIPV, is a highly fatal disease of cats with no currently available vaccine or US Food and Drug Administration-approved cure. Dissemination of FIPV in affected cats results in a range of clinical signs, including cavitary effusions, anorexia, fever and lesions of pyogranulomatous vasculitis and perivasculitis, with or without central nervous system or ocular involvement. The objectives of this study were to screen an array of antiviral compounds for anti-FIPV (serotype II) activity, determine cytotoxicity safety profiles of identified compounds with anti-FIPV activity and strategically combine identified monotherapies to assess compound synergy against FIPV in vitro. Based upon clinically successful combination treatment strategies for human patients with HIV and hepatitis C virus infections, we hypothesized that a combined anticoronaviral therapy approach featuring concurrent multiple mechanisms of drug action would result in an additive or synergistic antiviral effect. Methods This study screened 90 putative antiviral compounds for efficacy and cytotoxicity using a multimodal in vitro strategy, including plaque bioassays, real-time RT-PCR viral inhibition and cytotoxicity assays. Results Through this process, we identified 26 compounds with effective antiviral activity against FIPV, representing a variety of drug classes and mechanisms of antiviral action. The most effective compounds include GC376, GS-441524, EIDD2081 and EIDD2931. We documented antiviral efficacy for combinations of antiviral agents, with a few examined drug combinations demonstrating evidence of limited synergistic antiviral activity. Conclusions and relevance Although evidence of compound synergy was identified for several combinations of antiviral agents, monotherapies were ultimately determined to be the most effective in the inhibition of viral transcription.

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A rational approach to identifying effective combined anticoronaviral therapies against feline coronavirus

10.1101/2020.07.09.195016 ◽

2020 ◽

Author(s):

S.E. Cook ◽

H. Vogel ◽

D. Castillo ◽

M. Olsen ◽

N. Pedersen ◽

...

Keyword(s):

Antiviral Activity ◽

Viral Replication ◽

Clinical Signs ◽

Treatment Strategies ◽

Antiviral Effect ◽

Inhibitory Effect ◽

Rational Approach ◽

Neurological Involvement ◽

Ocular Involvement

AbstractFeline infectious peritonitis (FIP), caused by a genetic mutant of feline enteric coronavirus known as FIPV, is a highly fatal disease of cats with no currently available vaccine or FDA-approved cure. Dissemination of FIPV in affected cats results in a range of clinical signs including cavitary effusions, anorexia, fever and lesions of pyogranulomatous vasculitis and peri-vasculitis with or without central nervous system and/or ocular involvement. There is a critical need for effective and approved antiviral therapies against coronaviruses including FIPV and zoonotic coronaviruses such as SARS-CoV-2, the cause of COVID-19. With regards to SARS-CoV-2, preliminary evidence suggests that there may be potential clinical and pathological overlap with feline coronaviral disease including enteric and neurological involvement in some cases. We have screened 89 putative antiviral compounds and have identified 25 compounds with antiviral activity against FIPV, representing a variety of drug classes and mechanisms of antiviral action. Based upon successful combination treatment strategies for human patients with HIV or hepatitis C virus infections, we have identified combinations of drugs targeting different steps of the FIPV life cycle resulting in synergistic antiviral effect. Translationally, we suggest that a combined anticoronaviral therapy (cACT) with multiple mechanisms of action and penetration of all potential anatomic sites of viral infection should be applied towards other challenging to treat coronaviruses, like SARS-CoV-2.Author summaryWe have screened 89 compounds in vitro for antiviral activity against FIPV. The putative antiviral activity of these compounds was either purported to be a direct effect on viral proteins involved in viral replication or an indirect inhibitory effect on normal cellular pathways usurped by FIPV to aid viral replication. Twenty-five of these compounds were found to have significant antiviral activity. Certain combinations of these compounds were determined to be superior to monotherapy alone.

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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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Novel Nucleoside Analogues as Effective Antiviral Agents for Zika Virus Infections

Molecules ◽

10.3390/molecules25204813 ◽

2020 ◽

Vol 25 (20) ◽

pp. 4813

Author(s):

Marcella Bassetto ◽

Cecilia M. Cima ◽

Mattia Basso ◽

Martina Salerno ◽

Frank Schwarze ◽

...

Keyword(s):

Zika Virus ◽

Antiviral Agents ◽

Antiviral Agent ◽

Antiviral Effect ◽

Health Concern ◽

Virus Infections ◽

Direct Acting Antiviral ◽

Direct Acting ◽

Further Development

Previously considered a neglected flavivirus, Zika virus has recently emerged as a public health concern due to its ability to spread rapidly and cause severe neurological disorders, such as microcephaly in newborn babies from infected mothers, and Guillain-Barré syndrome in adults. Despite extensive efforts towards the identification of effective therapies, specific antivirals are still not available. As part of ongoing medicinal chemistry studies to identify new antiviral agents, we screened against Zika virus replication in vitro in a targeted internal library of small-molecule agents, comprising both nucleoside and non-nucleoside agents. Among the compounds evaluated, novel aryloxyphosphoramidate prodrugs of the nucleosides 2′-C-methyl-adenosine, 2-CMA, and 7-deaza-2′C-methyl-adenosine, 7-DMA, were found to significantly inhibit the virus-induced cytopathic effect in multiple relevant cell lines. In addition, one of these prodrugs exhibits a synergistic antiviral effect against Zika virus when applied in combination with an indirect antiviral agent, a l-dideoxy bicyclic pyrimidine nucleoside analogue, which potently inhibits vaccinia and measles viruses in vitro by targeting a host pathway. Our findings provide a solid basis for further development of an antiviral therapy for Zika virus infections, possibly exploiting a dual approach combining two different agents, one targeting the viral polymerase (direct-acting antiviral), the second targeting a host-directed autophagy mechanism.

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EGYVIR: An immunomodulatory herbal extract with potent antiviral activity against SARS-CoV-2

PLoS ONE ◽

10.1371/journal.pone.0241739 ◽

2020 ◽

Vol 15 (11) ◽

pp. e0241739

Author(s):

Wael H. Roshdy ◽

Helmy A. Rashed ◽

Ahmed Kandeil ◽

Ahmed Mostafa ◽

Yassmin Moatasim ◽

...

Keyword(s):

Antiviral Activity ◽

Nuclear Translocation ◽

Antiviral Agents ◽

In Vitro Study ◽

Antiviral Effect ◽

Black Pepper ◽

Infection Process ◽

Herbal Extract

Due to the challenges for developing vaccines in devastating pandemic situations of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), developing and screening of novel antiviral agents are peremptorily demanded. Herein, we developed EGYVIR as a potent immunomodulatory herbal extract with promising antiviral activity against SARS-CoV-2. It constitutes of a combination of black pepper extract with curcumin extract. The antiviral effect of EGYVIR extract is attributed to the two key phases of the disease in severe cases. First, the inhibition of the nuclear translocation of NF-kβ p50, attenuating the SARS-CoV-2 infection-associated cytokine storm. Additionally, the EGYVIR extract has an in vitro virucidal effect for SARS-CoV-2. The in vitro study of EGYVIR extract against SARS-CoV-2 on Huh-7 cell lines, revealed the potential role of NF-kβ/TNFα/IL-6 during the infection process. EGYVIR antagonizes the NF-kβ pathway in-silico and in-vitro studies. Consequently, it has the potential to hinder the release of IL-6 and TNFα, decreasing the production of essential cytokines storm elements.

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The Combined Use of Alphavirus Replicons and Pseudoinfectious Particles for the Discovery of Antivirals Derived from Natural Products

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057114564868 ◽

2014 ◽

Vol 20 (5) ◽

pp. 673-680 ◽

Cited By ~ 4

Author(s):

Phillip C. Delekta ◽

Avi Raveh ◽

Martha J. Larsen ◽

Pamela J. Schultz ◽

Giselle Tamayo-Castillo ◽

...

Keyword(s):

Life Cycle ◽

Antiviral Activity ◽

Viral Entry ◽

Particle System ◽

Antiviral Agents ◽

Viral Life Cycle ◽

Genome Replication ◽

Bacterial Strains ◽

Antiviral Compounds ◽

Western Equine Encephalitis

Alphaviruses are a prominent class of reemergent pathogens due to their globally expanding ranges, potential for lethality, and possible use as bioweapons. The absence of effective treatments for alphaviruses highlights the need for innovative strategies to identify antiviral agents. Primary screens that use noninfectious self-replicating RNAs, termed replicons, have been used to identify potential antiviral compounds for alphaviruses. Only inhibitors of viral genome replication, however, will be identified using replicons, which excludes many other druggable steps in the viral life cycle. To address this limitation, we developed a western equine encephalitis virus pseudoinfectious particle system that reproduces several crucial viral life cycle steps in addition to genome replication. We used this system to screen a library containing ~26,000 extracts derived from marine microbes, and we identified multiple bacterial strains that produce compounds with potential antiviral activity. We subsequently used pseudoinfectious particle and replicon assays in parallel to counterscreen candidate extracts, and followed antiviral activity during biochemical fractionation and purification to differentiate between inhibitors of viral entry and genome replication. This novel process led to the isolation of a known alphavirus entry inhibitor, bafilomycin, thereby validating the approach for the screening and identification of potential antiviral compounds.

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The in vitro and in vivo potency of CT-P59 against Delta and its associated variants of SARS-CoV-2

10.1101/2021.07.23.453472 ◽

2021 ◽

Author(s):

Dong-Kyun Ryu ◽

Hye-Min Woo ◽

Bobin Kang ◽

Hanmi Noh ◽

Jong-In Kim ◽

...

Keyword(s):

Antiviral Activity ◽

Respiratory Tract ◽

Animal Studies ◽

Antiviral Effect ◽

The World ◽

Symptom Remission ◽

Challenge Experiment ◽

Reduced Activity

The Delta variant originally from India is rapidly spreading across the world and causes to resurge infections of SARS-CoV-2. We previously reported that CT-P59 presented its in vivo potency against Beta and Gamma variants, despite its reduced activity in cell experiments. Yet, it remains uncertain to exert the antiviral effect of CT-P59 on the Delta and its associated variants (L452R). To tackle this question, we carried out cell tests and animal study. CT-P59 showed reduced antiviral activity but enabled neutralization against Delta, Epsilon, and Kappa variants in cells. In line with in vitro results, the mouse challenge experiment with the Delta variant substantiated in vivo potency of CT-P59 showing symptom remission and virus abrogation in the respiratory tract. Collectively, cell and animal studies showed that CT-P59 is effective against the Delta variant infection, hinting that CT-P59 has therapeutic potency for patients infected with Delta and its associated variants.

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Hydroxychloroquine in Patients with Rheumatic Disease Complicated by COVID-19: Clarifying Target Exposures and the Need for Clinical Trials

The Journal of Rheumatology ◽

10.3899/jrheum.200493 ◽

2020 ◽

Vol 47 (9) ◽

pp. 1424-1430 ◽

Cited By ~ 5

Author(s):

Stephen J. Balevic ◽

Christoph P. Hornik ◽

Thomas P. Green ◽

Megan E.B. Clowse ◽

Daniel Gonzalez ◽

...

Keyword(s):

Clinical Trials ◽

Antiviral Activity ◽

Rheumatic Disease ◽

Rheumatic Diseases ◽

Pharmacokinetic Model ◽

Therapeutic Window ◽

Total Serum ◽

Viral Inhibition ◽

Target Exposure

Objective.To characterize hydroxychloroquine (HCQ) exposure in patients with rheumatic disease receiving longterm HCQ compared to target concentrations with reported antiviral activity against the coronavirus disease 2019 caused by SARS-CoV-2 (COVID-19).Method.We evaluated total HCQ concentrations in serum and plasma from published literature values, frozen serum samples from a pediatric systemic lupus erythematosus trial, and simulated concentrations using a published pharmacokinetic model during pregnancy. For each source, we compared observed or predicted HCQ concentrations to target concentrations with reported antiviral activity against SARS-CoV-2.Results.The average total serum/plasma HCQ concentrations were below the lowest SARS-CoV-2 target of 0.48 mg/l in all studies. Assuming the highest antiviral target exposure (total plasma concentration of 4.1 mg/l), all studies had about one-tenth the necessary concentration for in vitro viral inhibition. Pharmacokinetic model simulations confirmed that pregnant adults receiving common dosing for rheumatic diseases did not achieve target exposures; however, the models predict that a dosage of 600 mg once a day during pregnancy would obtain the lowest median target exposure for most patients after the first dose.Conclusion.We found that the average patient receiving treatment with HCQ for rheumatic diseases, including children and non-pregnant/pregnant adults, are unlikely to achieve total serum or plasma concentrations shown to inhibit SARS-CoV-2 in vitro. Nevertheless, patients receiving HCQ long term may have tissue concentrations far exceeding that of serum/plasma. Because the therapeutic window for HCQ in the setting of SARS-CoV-2 is unknown, well-designed clinical trials that include patients with rheumatic disease are urgently needed to characterize the efficacy, safety, and target exposures for HCQ.

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Potent In Vivo Antiviral Activity of the Herpes Simplex Virus Primase-Helicase Inhibitor BAY 57-1293

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.46.6.1766-1772.2002 ◽

2002 ◽

Vol 46 (6) ◽

pp. 1766-1772 ◽

Cited By ~ 87

Author(s):

Ulrich A. K. Betz ◽

Rüdiger Fischer ◽

Gerald Kleymann ◽

Martin Hendrix ◽

Helga Rübsamen-Waigmann

Keyword(s):

Herpes Simplex Virus ◽

Herpes Simplex ◽

Antiviral Activity ◽

Lethal Challenge ◽

Antiviral Compounds ◽

Ocular Herpes ◽

Spread Model ◽

Simplex Virus

ABSTRACT BAY 57-1293 belongs to a new class of antiviral compounds and inhibits replication of herpes simplex virus (HSV) type 1 and type 2 in the nanomolar range in vitro by abrogating the enzymatic activity of the viral primase-helicase complex. In various rodent models of HSV infection the antiviral activity of BAY 57-1293 in vivo was found to be superior compared to all compounds currently used to treat HSV infections. The compound shows profound antiviral activity in murine and rat lethal challenge models of disseminated herpes, in a murine zosteriform spread model of cutaneous disease, and in a murine ocular herpes model. It is active in parenteral, oral, and topical formulations. BAY 57-1293 continued to demonstrate efficacy when the onset of treatment was initiated after symptoms of herpetic disease were already apparent.

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Studies on antiviral agents. III. Synthesis and in vitro antiviral activity of 1-N-higher-acyl-3"-N-functionalized acylkanamycin A derivatives.

The Journal of Antibiotics ◽

10.7164/antibiotics.38.1719 ◽

1985 ◽

Vol 38 (12) ◽

pp. 1719-1737 ◽

Cited By ~ 2

Author(s):

KEIJI MATSUDA ◽

NOBUYOSHI YASUDA ◽

HIDEO TSUTSUMI ◽

TAKAO TAKAYA

Keyword(s):

Antiviral Activity ◽

Antiviral Agents

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Nilotinib Exhibits an in Vitro Antiviral Activity Against Human Cytomegalovirus (HCMV): Potential Clinical Applications

Blood ◽

10.1182/blood.v120.21.4666.4666 ◽

2012 ◽

Vol 120 (21) ◽

pp. 4666-4666 ◽

Cited By ~ 2

Author(s):

Dana Wolf ◽

Esther Djian ◽

Katia Beider ◽

Avichai Shimoni ◽

Arnon Nagler

Keyword(s):

Antiviral Activity ◽

Antiviral Therapy ◽

Conflicts Of Interest ◽

Philadelphia Chromosome ◽

Chronic Phase ◽

Host Cells ◽

Viral Inhibition ◽

Advanced Phase ◽

Hcmv Disease

Abstract Abstract 4666 Chronic myeloid leukemia (CML) is a clonal disorder associated with chromosomal translocation t (9; 22), which produces the Philadelphia chromosome. The fusion gene encodes for the chimeric oncoprotein BCR-ABL, associated with deregulated constitutive tyrosine kinase (TK) activity, leading to leukemogenesis. Imatinib mesylate, the first potent selective inhibitor of BCR-ABL TK, has revolutionized the current management of CML. The second generation TK inhibitor (TKI) Nilotinib (Novartis, East Hanover, NJ) is an aminopyrimidine derivative of imatinib with an increased binding affinity to the chimeric p210 BCR-ABL. Nilotinib is active in imatinib-resistant or -intolerant patients in chronic phase, accelerated phase (AP), and blast crisis (BC) CML. Allogeneic stem cell transplantation (SCT) is currently reserved for patients(pts) in CP after the failure of second line TKI or for patients in advanced phase disease.SCT remains the treatment of choice in patients with Ph+ALL. TKIs can be used successfully pre -SCT as a bridge to SCT in advanced disease and post-SCT in order to prevent disease recurrence. HCMV is one of the leading causes of morbidity and mortality post SCT in particular in pts transplanted for advance CML and pts with graft vs. host disease (GVHD) While preemptive antiviral therapy has reduced the occurrence of HCMV disease post SCT, the use of all currently available antiviral drugs is often limited by toxicity, low oral bioavailability, and drug resistance. The mechanisms facilitating HCMV entry into the host cells are not clearly understood. Blocking of the platelet – derived growth factor receptor-α (PDGFRα) has been shown to inhibit HCMV internalization and gene expression. Recently, Imatinib have been shown to inhibit PDGFRα phosphorylation. As Nilotinib is a more potent PDGFR inhibitor, we assessed the in vitro antiviral activity of Nilotinib against HCMV. Nilotinib exhibited a significant dose-dependent inhibition of the virus upon pre-incubation with the drug. Moreover, Nilotinib demonstrated a ∼4.5-fold higher antiviral activity against HCMV when compared to imatinib, with IC50 values of 0.39 μM and 1.75 μM, respectively. No viral inhibition was found upon addition of the drugs after viral adsorption – compatible with inhibition of an early step of infection, involving viral binding/entry into the cell. These findings identify a promising new target for antiviral therapy, representing an alternative paradigm for treatment with compounds combining anti-cancer and antiviral activity. It remains to be determined if the anti-HCMV activity demonstrated for Nilotinib is of clinical relevance in patients undergoing SCT Disclosures: No relevant conflicts of interest to declare.

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