Discovery of clinically approved drugs capable of inhibiting SARS-CoV-2 in vitro infection using a phenotypic screening strategy and network-analysis to predict their potential to treat covid-19

AbstractThe disease caused by SARS-CoV2, covid-19, rapidly spreads worldwide, causing the greatest threat to global public health in the last 100 years. This scenario has become catastrophic as there are no approved vaccines to prevent the disease, and the main measures to contain the virus transmission are confinement and social distancing. One priority strategy is based on drug repurposing by pursuing antiviral chemotherapy that can control transmission and prevent complications associated with covid-19. With this aim, we performed a high content screening assay for the discovery of anti-SARS-CoV-2 compounds. From the 65 screened compounds, we have found four drugs capable to selectively inhibit SARS-CoV-2 in vitro infection: brequinar, abiraterone acetate, neomycin, and the extract of Hedera helix. Brequinar and abiraterone acetate had higher inhibition potency against SARS-CoV-2 than neomycin and Hedera helix extract, respectively. Drugs with reported antiviral activity and in clinical trials for covid-19, chloroquine, ivermectin, and nitazoxanide, were also included in the screening, and the last two were found to be non-selective. We used a data mining approach to build drug-host molecules-biological function-disease networks to show in a holistic way how each compound is interconnected with host node molecules and virus infection, replication, inflammatory response, and cell apoptosis. In summary, the present manuscript identified four drugs with active inhibition effect on SARS-CoV-2 in vitro infection, and by network analysis, we provided new insights and starting points for the clinical evaluation and repurposing process to treat SARS-CoV-2 infection.Summary sentenceDiscovery of drug repurposing candidates, inhibitors of SARS-CoV-2 infection in vitro, using a phenotypic screening strategy and network analysis.

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Antiparasitic Properties of Cardiovascular Agents against Human Intravascular Parasite Schistosoma mansoni

Pharmaceuticals ◽

10.3390/ph14070686 ◽

2021 ◽

Vol 14 (7) ◽

pp. 686

Author(s):

Raquel Porto ◽

Ana C. Mengarda ◽

Rayssa A. Cajas ◽

Maria C. Salvadori ◽

Fernanda S. Teixeira ◽

...

Keyword(s):

Schistosoma Mansoni ◽

Screening Strategy ◽

Early Infection ◽

Global Public Health ◽

Parasitic Worm ◽

Electron Microscopy Analysis ◽

Health Significance ◽

Microscopy Analysis

The intravascular parasitic worm Schistosoma mansoni is a causative agent of schistosomiasis, a disease of great global public health significance. Praziquantel is the only drug available to treat schistosomiasis and there is an urgent demand for new anthelmintic agents. Adopting a phenotypic drug screening strategy, here, we evaluated the antiparasitic properties of 46 commercially available cardiovascular drugs against S. mansoni. From these screenings, we found that amiodarone, telmisartan, propafenone, methyldopa, and doxazosin affected the viability of schistosomes in vitro, with effective concentrations of 50% (EC50) and 90% (EC90) values ranging from 8 to 50 µM. These results were further supported by scanning electron microscopy analysis. Subsequently, the most effective drug (amiodarone) was further tested in a murine model of schistosomiasis for both early and chronic S. mansoni infections using a single oral dose of 400 mg/kg or 100 mg/kg daily for five consecutive days. Amiodarone had a low efficacy in chronic infection, with the worm and egg burden reduction ranging from 10 to 30%. In contrast, amiodarone caused a significant reduction in worm and egg burden in early infection (>50%). Comparatively, treatment with amiodarone is more effective in early infection than praziquantel, demonstrating the potential role of this cardiovascular drug as an antischistosomal agent.

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Ebolavirus Entry: From Molecular Characterization to Drug Discovery

10.20944/preprints201902.0209.v1 ◽

2019 ◽

Cited By ~ 1

Author(s):

Cristiano Salata ◽

Arianna Calistri ◽

Gualtiero Alvisi ◽

Michele Celestino ◽

Cristina Parolin ◽

...

Keyword(s):

Life Cycle ◽

Viral Entry ◽

Large Scale ◽

Ebola Virus ◽

Virus Disease ◽

Drug Repurposing ◽

In Vitro Study ◽

Rational Drug Design ◽

Global Public Health

Ebola Virus Disease (EVD) is one of the most lethal transmissible infections characterized by a high fatality rate, and caused by members of the Filoviridae family. The recent large outbreak of EVD in West Africa (2013-2016), highlighted the worldwide danger of this disease and its impact on global public health and economy. The development of highly needed anti-Filoviridae antivirals has been so far hampered by the shortage of tools to study their life cycle in vitro, and therefore screen for potential active compounds outside a biosafety level-4 (BSL-4) containment. Importantly, the development of surrogate models to in vitro study of Filoviridae entry in a BSL-2 setting, such as viral pseudotypes and Ebola virus like particles, tremendously boosted both our knowledge on viral life cycle and the identification of promising anti-Filoviridae compounds interfering with viral entry. In this context, the combination of such surrogate systems with large-scale small molecule compounds and haploid genetic screenings, as well as rational drug design and drug repurposing approaches will prove priceless in our quest for the development of a treatment for EVD.

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Ebola Virus Entry: From Molecular Characterization to Drug Discovery

Viruses ◽

10.3390/v11030274 ◽

2019 ◽

Vol 11 (3) ◽

pp. 274 ◽

Cited By ~ 16

Author(s):

Cristiano Salata ◽

Arianna Calistri ◽

Gualtiero Alvisi ◽

Michele Celestino ◽

Cristina Parolin ◽

...

Keyword(s):

Life Cycle ◽

Viral Entry ◽

Large Scale ◽

Ebola Virus ◽

Virus Disease ◽

Virus Entry ◽

Drug Repurposing ◽

Rational Drug Design ◽

Global Public Health

Ebola Virus Disease (EVD) is one of the most lethal transmissible infections, characterized by a high fatality rate, and caused by a member of the Filoviridae family. The recent large outbreak of EVD in Western Africa (2013–2016) highlighted the worldwide threat represented by the disease and its impact on global public health and the economy. The development of highly needed anti-Ebola virus antivirals has been so far hampered by the shortage of tools to study their life cycle in vitro, allowing to screen for potential active compounds outside a biosafety level-4 (BSL-4) containment. Importantly, the development of surrogate models to study Ebola virus entry in a BSL-2 setting, such as viral pseudotypes and Ebola virus-like particles, tremendously boosted both our knowledge of the viral life cycle and the identification of promising antiviral compounds interfering with viral entry. In this context, the combination of such surrogate systems with large-scale small molecule compounds and haploid genetic screenings, as well as rational drug design and drug repurposing approaches will prove priceless in our quest for the development of a treatment for EVD.

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Pediatric Drug Nitazoxanide: A Potential Choice for Control of Zika

Open Forum Infectious Diseases ◽

10.1093/ofid/ofx009 ◽

2017 ◽

Vol 4 (1) ◽

Cited By ~ 24

Author(s):

Rui-Yuan Cao ◽

Yong-fen Xu ◽

Tian-Hong Zhang ◽

Jing-Jing Yang ◽

Ye Yuan ◽

...

Keyword(s):

Zika Virus ◽

Congenital Malformations ◽

Drug Repurposing ◽

Antiviral Effect ◽

Public Health Emergency ◽

Global Public Health ◽

Pediatric Drug ◽

Zikv Infection ◽

Pediatric Use

Abstract Zika virus (ZIKV) infection can be the cause of congenital malformations, including microcephaly in infants and can cause other disorders such as Guillain-Barré syndrome, meningoencephalitis, and myelitis, which can also occur in some infected adults. However, at this time, there is no drug approved to treat ZIKV infection. Drug repurposing is the promptest way to obtain an effective drug during a global public health emergency such as the spread of Zika virus. In this study, we report a US Food and Drug Admistration-approved drug that is safe for pediatric use. Nitazoxanide and its bioactive metabolite, tizoxanide, have anti-ZIKV potential in vitro, and we identified that they exerts antiviral effect possibly by targeting the viral postattachment step.

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477 Deep learning to drive COVID-19 rapid drug repurposing

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2020-sitc2020.0477 ◽

2020 ◽

Vol 8 (Suppl 3) ◽

pp. A509-A509

Author(s):

Sarah Kolitz ◽

Jason Kim ◽

Jenny Zhang ◽

Yoonjeong Cha ◽

Sailaja Battula ◽

...

Keyword(s):

Deep Learning ◽

Validation Studies ◽

Plaque Assay ◽

Drug Repurposing ◽

Drug Binding ◽

Global Public Health ◽

Viral Inhibition ◽

Health Crisis ◽

Oak Ridge

BackgroundCOVID-19 is a global public health crisis with no effective therapeutic strategies or vaccines available. The disease is caused by the SARS-CoV-2 virus, a novel coronavirus that enters cells through the ACE2 receptor. To rapidly identify existing drugs that might preferentially bind to the ACE2 receptor we sought to use an artificial intelligence platform to evaluate ~3,000 known drugs in the FDA approved drug library (Selleckchem).MethodsFluency is a quantitative structure–activity relationship (QSAR) deep learning-based platform that evaluates small molecule drug binding to protein targets. All drug structures from the FDA approved library were evaluated for binding to the ACE2 receptor and re-filtered for preferential ACE2 vs. ACE1 receptor binding. Top hits were evaluated for specificity by predicting binding across the human proteome and filtered by evaluating rankings from each of two models along with average ranks and combined scores from both models. The drugs were then evaluated for classification, potential availability and prioritized for in vitro validation. Selected compounds were screened using a high-throughput SARS-CoV-2 cell-based assay as described previously (Jonsson et al. J Biomol Screen 2007 12: 33. DOI: 10.1177/1087057106296688). Plates are quality-controlled in each run using Z score and CV statistics. Positive controls consisting of cells only and negative controls consisting of virus were used to normalize the data. Individual drugs are added to each plate at a single dose with at least four doses tested. For titer reduction assays, VeroE6 cells are infected with virus at MOI of 0.1 for one hour to promote adsorption. After two days, the supernatant is harvested and the amount of virus in each well is measured using TCID50 or plaque assay.ResultsWe identified 25 top drugs that were predicted to bind to ACE2 receptors and could theoretically block SARS-CoV-2 cell entry. Of these drugs, we prioritized 12 drugs for validation covering multiple pharmacologic classes and after assessing drug availability (table 1). They included an ALK/EGFR inhibitor, JAK inhibitor, two electrolyte channel inhibitors, \an antibiotic, and several anti-viral drugs, ACE inhibitors and anticoagulants. Validation studies are in progress and viral inhibition and titer reduction data will be presented.ConclusionsOur data show that machine learning platforms can be used to rapidly identify existing drugs that may have activity against SARS-CoV-2 infection. This hybrid computational and experimental approach enables rapid discovery of drugs for clinical testing against COVID-19 and other emerging human diseases.AcknowledgementsWe would like to thank Dr. Colleen Jonsson and Dr. Jeremy Smith at Oak Ridge National Laboratories and the University of Tennessee Regional Biocontainment Laboratory for assistance with in vitro validation studies.Trial RegistrationN/AEthics ApprovalN/AConsentN.A

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Evaluation of broad-spectrum antiviral compounds against chikungunya infection using a phenotypic screening strategy

F1000Research ◽

10.12688/f1000research.16498.1 ◽

2018 ◽

Vol 7 ◽

pp. 1730 ◽

Cited By ~ 1

Author(s):

Rafaela M. Bonotto ◽

Glaucia Souza-Almeida ◽

Soraya Jabur Badra ◽

Luiz Tadeu Figueiredo ◽

Carolina B. Moraes ◽

...

Keyword(s):

Broad Spectrum ◽

Treatment Options ◽

Public Health Problem ◽

Drug Repurposing ◽

Drug Approval ◽

Cell Number ◽

Phenotypic Screening ◽

Screening Assay ◽

Additional Advantage ◽

Antiviral Compounds

Chikungunya fever is an emerging disease and a significant public health problem in tropical countries. Recently reported outbreaks in Brazil in 2015 drew attention to the need to develop prevention and treatment options, as no antiviral chemotherapy or vaccines are currently available for this disease. Two strategies have been proved to accelerate the discovery of new anti-infectives: phenotypic screening and drug repurposing. Phenotypic screening can support the fast interrogation of compounds without the need for a pre-validated drug target, which is not available for the chikungunya virus (CHIKV) and has the additional advantage of facilitating the discovery of antiviral with novel mechanism of action. Drug repurposing can save time and resources in drug development by enabling secondary uses for drugs that are already approved for human treatment, thus precluding the need for several of the mandatory preclinical and clinical studies necessary for drug approval. A phenotypic screening assay was developed by infecting the human hepatoma Huh-7 cells with CHIKV 181/25 and quantifying infection through indirect immunofluorescence. The compound 6-azauridine was used as a positive control drug. The screening assay was validated by testing a commercial library of 1,280 compounds, including FDA-approved drugs, and used to screen a panel of broad-spectrum antiviral compounds for anti-CHIKV activity. A high content assay was set up in Huh-7 cells-infected with CHIKV. The maximum rate of infection peaked at 48 hours post-infection, after which the host cell number was greatly reduced due to a strong cytopathic effect. Assay robustness was confirmed with Z’-factor values >0.8 and high correlation coefficient between independent runs, demonstrating that the assay is reliable, consistent and reproducible. Among tested compounds, sofosbuvir, an anti-hepatitis C virus drug, exhibited good selectivity against CHIKV with an EC50of 11 µM, suggesting it is a promising candidate for repurposing.

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In Vitro Screening of Azalea for Resistance to Azalea Lace Bug

HortScience ◽

10.21273/hortsci.33.3.506b ◽

1998 ◽

Vol 33 (3) ◽

pp. 506b-506

Author(s):

Carol D. Robacker ◽

S.K. Braman

Keyword(s):

Leaf Damage ◽

In Vitro Screening ◽

Screening Assay ◽

Delaware Valley ◽

In Vitro Techniques ◽

Culture Tube ◽

Lace Bug ◽

Lace Bugs ◽

Laboratory Bioassays

Azalea lace bug (Stephanitis pyrioides) is the most serious pest on azalea. Results of laboratory bioassays and field evaluations of 17 deciduous azalea taxa have identified three resistant taxa: R. canescens, R. periclymenoides, and R. prunifolium. Highly susceptible taxa are `Buttercup', `My Mary', R. oblongifolium, and the evergreen cultivar `Delaware Valley White'. To determine whether in vitro techniques would have potential value in screening or selecting for resistance, or for the identification of morphological or chemical factors related to resistance, an in-vitro screening assay was developed. In-vitro shoot proliferation was obtained using the medium and procedures of Economou and Read (1984). Shoots used in the bioassays were grown in culture tubes. Two assays were developed: one for nymphs and one for adult lace bugs. To assay for resistance to nymphs, `Delaware Valley White' leaves containing lace bug eggs were disinfested with 70% alcohol and 20% commercial bleach, and incubated in sterile petri plates with moistened filter paper until the nymphs hatched. Five nymphs were placed in each culture tube, and cultures were incubated for about 2 weeks, or until adults were observed. To assay for resistance to adults, five female lace bugs were placed in each culture tube and allowed to feed for 5 days. Data collected on survival and leaf damage was generally supportive of laboratory bioassays and field results. Adult lace bugs had a low rate of survival on resistant taxa. Survival of nymphs was somewhat reduced on resistant taxa.

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The Antiviral and Antimalarial Drug Repurposing in Quest of Chemotherapeutics to Combat COVID-19 Utilizing Structure-Based Molecular Docking

Combinatorial Chemistry & High Throughput Screening ◽

10.2174/1386207323999200824115536 ◽

2020 ◽

Vol 23 ◽

Author(s):

Sisir Nandi ◽

Mohit Kumar ◽

Mridula Saxena ◽

Anil Kumar Saxena

Keyword(s):

Molecular Docking ◽

In Silico ◽

Drug Repurposing ◽

Clinical Settings ◽

The Novel ◽

In Silico Docking ◽

Biochemical Mechanisms ◽

Novel Coronavirus ◽

In Silico Molecular Docking

Background: The novel coronavirus disease (COVID-19) is caused by a new strain (SARS-CoV-2) erupted in 2019. Nowadays, it is a great threat that claims uncountable lives worldwide. There is no specific chemotherapeutics developed yet to combat COVID-19. Therefore, scientists have been devoted in the quest of the medicine that can cure COVID- 19. Objective: Existing antivirals such as ASC09/ritonavir, lopinavir/ritonavir with or without umifenovir in combination with antimalarial chloroquine or hydroxychloroquine have been repurposed to fight the current coronavirus epidemic. But exact biochemical mechanisms of these drugs towards COVID-19 have not been discovered to date. Method: In-silico molecular docking can predict the mode of binding to sort out the existing chemotherapeutics having a potential affinity towards inhibition of the COVID-19 target. An attempt has been made in the present work to carry out docking analyses of 34 drugs including antivirals and antimalarials to explain explicitly the mode of interactions of these ligands towards the COVID-19protease target. Results: 13 compounds having good binding affinity have been predicted towards protease binding inhibition of COVID-19. Conclusion: Our in silico docking results have been confirmed by current reports from clinical settings through the citation of suitable experimental in vitro data available in the published literature.

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Rhamnolipids Nano-Micelles as a Potential Hand Sanitizer

Antibiotics ◽

10.3390/antibiotics10070751 ◽

2021 ◽

Vol 10 (7) ◽

pp. 751

Author(s):

Marwa Reda Bakkar ◽

Ahmed Hassan Ibrahim Faraag ◽

Elham R. S. Soliman ◽

Manar S. Fouda ◽

Amir Mahfouz Mokhtar Sarguos ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Antimicrobial Activity ◽

Active Sites ◽

Specific Interaction ◽

Virus Transmission ◽

Multidrug Resistant ◽

Future Perspective ◽

Resistant Bacteria ◽

In Silico Studies

COVID-19 is a pandemic disease caused by the SARS-CoV-2, which continues to cause global health and economic problems since emerging in China in late 2019. Until now, there are no standard antiviral treatments. Thus, several strategies were adopted to minimize virus transmission, such as social distancing, face covering protection and hand hygiene. Rhamnolipids are glycolipids produced formally by Pseudomonas aeruginosa and as biosurfactants, they were shown to have broad antimicrobial activity. In this study, we investigated the antimicrobial activity of rhamnolipids against selected multidrug resistant bacteria and SARS-CoV-2. Rhamnolipids were produced by growing Pseudomonas aeruginosa strain LeS3 in a new medium formulated from chicken carcass soup. The isolated rhamnolipids were characterized for their molecular composition, formulated into nano-micelles, and the antibacterial activity of the nano-micelles was demonstrated in vitro against both Gram-negative and Gram-positive drug resistant bacteria. In silico studies docking rhamnolipids to structural and non-structural proteins of SARS-CoV-2 was also performed. We demonstrated the efficient and specific interaction of rhamnolipids with the active sites of these proteins. Additionally, the computational studies suggested that rhamnolipids have membrane permeability activity. Thus, the obtained results indicate that SARS-CoV-2 could be another target of rhamnolipids and could find utility in the fight against COVID-19, a future perspective to be considered.

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High-resolution AP-SMALDI MSI as a tool for drug imaging in Schistosoma mansoni

Analytical and Bioanalytical Chemistry ◽

10.1007/s00216-021-03230-w ◽

2021 ◽

Author(s):

Annika S. Mokosch ◽

Stefanie Gerbig ◽

Christoph G. Grevelding ◽

Simone Haeberlein ◽

Bernhard Spengler

Keyword(s):

Schistosoma Mansoni ◽

Egg Production ◽

Drug Repurposing ◽

Cancer Drug ◽

Ionization Mass ◽

Parasitic Flatworm ◽

Paired Female ◽

First Time ◽

Organ Tropism

AbstractSchistosoma mansoni is a parasitic flatworm causing schistosomiasis, an infectious disease affecting several hundred million people worldwide. Schistosomes live dioeciously, and upon pairing with the male, the female starts massive egg production, which causes pathology. Praziquantel (PZQ) is the only drug used, but it has an inherent risk of resistance development. Therefore, alternatives are needed. In the context of drug repurposing, the cancer drug imatinib was tested, showing high efficacy against S. mansoni in vitro. Besides the gonads, imatinib mainly affected the integrity of the intestine in males and females. In this study, we investigated the potential uptake and distribution of imatinib in adult schistosomes including its distribution kinetics. To this end, we applied for the first time atmospheric-pressure scanning microprobe matrix-assisted laser desorption/ionization mass spectrometry imaging (AP-SMALDI MSI) for drug imaging in paired S. mansoni. Our results indicate that imatinib was present in the esophagus and intestine of the male as early as 20 min after in vitro exposure, suggesting an oral uptake route. After one hour, the drug was also found inside the paired female. The detection of the main metabolite, N-desmethyl imatinib, indicated metabolization of the drug. Additionally, a marker signal for the female ovary was successfully applied to facilitate further conclusions regarding organ tropism of imatinib. Our results demonstrate that AP-SMALDI MSI is a useful method to study the uptake, tissue distribution, and metabolization of imatinib in S. mansoni. The results suggest using AP-SMALDI MSI also for investigating other antiparasitic compounds and their metabolites in schistosomes and other parasites. Graphical abstract

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