High-throughput Antiviral Assays to Screen for Inhibitors of Zika Virus Replication

Fernandes, R.S.; Noske, G.D.; Gawriljuk, V.O.; Oliveira, Ketllyn Irene Zagato de; Godoy, Andre S.; Mesquita, Nathalya C. M. R.; Oliva, Glaucius

doi:10.3791/62422

Cited by 3 publications

(5 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…First, we evaluated antiprotease activity at a single time point using a single concentration of each selected compound (200 μM). The positive control aprotinin was used and had an IC 50 of 0.13 ± 0.02 μM . We observed that compounds LabMol-301 and LabMol-314 inhibited 86% and 83% of the protease activity, respectively.…”

Section: Resultsmentioning

confidence: 99%

“…NS5 RdRp and NS2B-NS3pro activity assays were performed as described by Fernandes and co-workers . To evaluate the inhibitory activity of the compounds against ZIKV NS5 RdRp, we performed enzymatic assays, previously described by Sáez-Álvarez and co-workers .…”

Section: Methodsmentioning

confidence: 99%

“…NS5 RdRp and NS2B-NS3pro activity assays were performed as described by Fernandes and co-workers. 92 To evaluate the inhibitory activity of the compounds against ZIKV NS5 RdRp, we performed enzymatic assays, previously described by Saéz-A ́lvarez and co-workers. 93 The concentration−response experiments for NS5 RdRp were performed with serial dilution of each compound ranging from 80 to 0.039 μM.…”

Section: ■ Introductionmentioning

confidence: 99%

“…94 To investigate the mechanism of inhibition of LabMol-301 against the ZIKV NS2B-NS3 protease, we used the fluorescence of the synthetic fluorogenic peptide benzoyl-Nle-Lys-Arg-Arg-4methylcoumarin-7-amide (Bz-nKRR-AMC) to determine the initial velocity of the enzymatic reactions in the presence of varying concentrations of the inhibitor. Kinetic assays were performed for LabMol-301 at 37 °C, using the protocol described by Fernandes and co-workers: 92 4 nM of protein, 50−3.1 μM of substrate Bz-nKRR-AMC, and 3.75 and 7.5 μM of LabMol-301. Enzymatic assays were performed in duplicate on a 96-well flat plate and in a SpectraMax Gemini EM microplate reader (Molecular Devices Co.).…”

Section: ■ Introductionmentioning

confidence: 99%

“…The positive control aprotinin was used and had an IC 50 of 0.13 ± 0.02 μM. 92 We observed that compounds LabMol-301 and LabMol-314 inhibited 86% and 83% of the protease activity, respectively. Therefore, these compounds were selected for concentration−response experiments to determine the IC 50 values.…”

Section: ■ Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Discovery of New Zika Protease and Polymerase Inhibitors through the Open Science Collaboration Project OpenZika

Mottin

Sousa

Mesquita

et al. 2022

J. Chem. Inf. Model.

Self Cite

View full text Add to dashboard Cite

The Zika virus (ZIKV) is a neurotropic arbovirus considered a global threat to public health. Although there have been several efforts in drug discovery projects for ZIKV in recent years, there are still no antiviral drugs approved to date. Here, we describe the results of a global collaborative crowdsourced open science project, the OpenZika project, from IBM's World Community Grid (WCG), which integrates different computational and experimental strategies for advancing a drug candidate for ZIKV. Initially, molecular docking protocols were developed to identify potential inhibitors of ZIKV NS5 RNA-dependent RNA polymerase (NS5 RdRp), NS3 protease (NS2B-NS3pro), and NS3 helicase (NS3hel). Then, a machine learning (ML) model was built to distinguish active vs inactive compounds for the cytoprotective effect against ZIKV infection. We performed three independent target-based virtual screening campaigns (NS5 RdRp, NS2B-NS3pro, and NS3hel), followed by predictions by the ML model and other filters, and prioritized a total of 61 compounds for further testing in enzymatic and phenotypic assays. This yielded five non-nucleoside compounds which showed inhibitory activity against ZIKV NS5 RdRp in enzymatic assays (IC 50 range from 0.61 to 17 μM). Two compounds thermally destabilized NS3hel and showed binding affinity in the micromolar range (K d range from 9 to 35 μM). Moreover, the compounds LabMol-301 inhibited both NS5 RdRp and NS2B-NS3pro (IC 50 of 0.8 and 7.4 μM, respectively) and LabMol-212 thermally destabilized the ZIKV NS3hel (K d of 35 μM). Both also protected cells from death induced by ZIKV infection in in vitro cell-based assays. However, while eight compounds (including LabMol-301 and LabMol-212) showed a cytoprotective effect and prevented ZIKV-induced cell death, agreeing with our ML model for prediction of this cytoprotective effect, no compound showed a direct antiviral effect against ZIKV. Thus, the new scaffolds discovered here are promising hits for future structural optimization and for advancing the discovery of further drug candidates for ZIKV. Furthermore, this work has demonstrated the importance of the integration of computational and experimental approaches, as well as the potential of large-scale collaborative networks to advance drug discovery projects for neglected diseases and emerging viruses, despite the lack of available direct antiviral activity and cytoprotective effect data, that reflects on the assertiveness of the computational predictions. The importance of these efforts rests with the need to be prepared for future viral epidemic and pandemic outbreaks.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Discovery of New Zika Protease and Polymerase Inhibitors through the Open Science Collaboration Project OpenZika

Mottin

Sousa

Mesquita

et al. 2022

J. Chem. Inf. Model.

Self Cite

View full text Add to dashboard Cite

show abstract

Chalcones from Angelica keiskei (ashitaba) inhibit key Zika virus replication proteins

Mottin

Caesar

Brodsky

et al. 2022

Bioorganic Chemistry

View full text Add to dashboard Cite

Natural Compounds as Non-Nucleoside Inhibitors of Zika Virus Polymerase through Integration of In Silico and In Vitro Approaches

et al. 2022

View full text Add to dashboard Cite

Although the past epidemic of Zika virus (ZIKV) resulted in severe neurological consequences for infected infants and adults, there are still no approved drugs to treat ZIKV infection. In this study, we applied computational approaches to screen an in-house database of 77 natural and semi-synthetic compounds against ZIKV NS5 RNA-dependent RNA-polymerase (NS5 RdRp), an essential protein for viral RNA elongation during the replication process. For this purpose, we integrated computational approaches such as binding-site conservation, chemical space analysis and molecular docking. As a result, we prioritized nine virtual hits for experimental evaluation. Enzymatic assays confirmed that pedalitin and quercetin inhibited ZIKV NS5 RdRp with IC50 values of 4.1 and 0.5 µM, respectively. Moreover, pedalitin also displayed antiviral activity on ZIKV infection with an EC50 of 19.28 µM cell-based assays, with low toxicity in Vero cells (CC50 = 83.66 µM) and selectivity index of 4.34. These results demonstrate the potential of the natural compounds pedalitin and quercetin as candidates for structural optimization studies towards the discovery of new anti-ZIKV drug candidates.

show abstract

High-throughput Antiviral Assays to Screen for Inhibitors of Zika Virus Replication

Cited by 3 publications

References 0 publications

Discovery of New Zika Protease and Polymerase Inhibitors through the Open Science Collaboration Project OpenZika

Discovery of New Zika Protease and Polymerase Inhibitors through the Open Science Collaboration Project OpenZika

Chalcones from Angelica keiskei (ashitaba) inhibit key Zika virus replication proteins

Natural Compounds as Non-Nucleoside Inhibitors of Zika Virus Polymerase through Integration of In Silico and In Vitro Approaches

Contact Info

Product

Resources

About