Cell-Based Antiviral Assays for Screening and Profiling Inhibitors Against Dengue Virus

Gong, Edwin Yunhao; Clynhens, Marleen; Ivens, Tania; Lory, Pedro; Simmen, Kenny; Kraus, Guenter

doi:10.1007/978-1-62703-484-5_15

Cited by 4 publications

(3 citation statements)

References 3 publications

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“…Inhibition values for antiviral molecules were plotted using KaleidaGraph plotting software (v.4.03, Synergy Software). The antiviral activity of JNJ-1802 against DENV-1/45AZ5 was tested in Vero cells by measuring viral RNA using RT–qPCR as described previously 56 . Antiviral activity/toxicity assays of JNJ-1802 for all other viruses, not belonging to the flaviviruses, were performed using either reporter systems, cell viability, RT–qPCR or plaque reduction assays as readout, as further specified in the Supplementary Information and Supplementary Table 2 .…”

Section: Methodsmentioning

confidence: 99%

Blocking NS3–NS4B interaction inhibits dengue virus in non-human primates

Goethals

Kaptein

Kesteleyn

et al. 2023

Nature

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Dengue is a major health threat and the number of symptomatic infections caused by the four dengue serotypes is estimated to be 96 million1 with annually around 10,000 deaths2. However, no antiviral drugs are available for the treatment or prophylaxis of dengue. We recently described the interaction between non-structural proteins NS3 and NS4B as a promising target for the development of pan-serotype dengue virus (DENV) inhibitors3. Here we present JNJ-1802—a highly potent DENV inhibitor that blocks the NS3–NS4B interaction within the viral replication complex. JNJ-1802 exerts picomolar to low nanomolar in vitro antiviral activity, a high barrier to resistance and potent in vivo efficacy in mice against infection with any of the four DENV serotypes. Finally, we demonstrate that the small-molecule inhibitor JNJ-1802 is highly effective against viral infection with DENV-1 or DENV-2 in non-human primates. JNJ-1802 has successfully completed a phase I first-in-human clinical study in healthy volunteers and was found to be safe and well tolerated4. These findings support the further clinical development of JNJ-1802, a first-in-class antiviral agent against dengue, which is now progressing in clinical studies for the prevention and treatment of dengue.

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Section: Methodsmentioning

confidence: 99%

Blocking NS3–NS4B interaction inhibits dengue virus in non-human primates

Goethals

Kaptein

Kesteleyn

et al. 2023

Nature

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“…Aliquots of the virus were stored at 80 • C. The 50% tissue culture infective dose (TCID50) was determined using the Promega Viral ToxGlo assay (Fisher Scientific, Ottawa, ON, Canada) in BHK-21 cells, according to the manufacturer's instructions. Cell-based assays for the evaluation of antiviral activity against DENV were performed as described previously [49,50] with modifications, using the Viral ToxGlo assay (Fisher Scientific, Ottawa, ON, Canada) according to the manufacturer's instructions. Briefly, BHK-21 cells were plated in 96-well plates at 2500 cells/well in Dulbecco's modified Eagle's medium (DMEM) containing 10% FBS and 1% penicillinstreptomycin.…”

Section: Antiviral Screening and Toxicity Assaysmentioning

confidence: 99%

Design, Synthesis, Evaluation and Molecular Dynamics Simulation of Dengue Virus NS5-RdRp Inhibitors

Zong,

Li,

et al. 2023

Pharmaceuticals

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Dengue virus (DENV) is a major mosquito-borne human pathogen in tropical countries; however, there are currently no targeted antiviral treatments for DENV infection. Compounds 27 and 29 have been reported to be allosteric inhibitors of DENV RdRp with potent inhibitory effects. In this study, the structures of compounds 27 and 29 were optimized using computer-aided drug design (CADD) approaches. Nine novel compounds were synthesized based on rational considerations, including molecular docking scores, free energy of binding to receptor proteins, predicted Absorption, Distribution, Metabolism, Excretion, and Toxicity (ADMET) parameters, structural diversity, and feasibility of synthesis. Subsequently, the anti-DENV activity was assessed. In the cytopathic effect (CPE) assay conducted on BHK-21 cells using the DENV2 NGC strain, both SW-b and SW-d demonstrated comparable or superior activity against DENV2, with IC50 values of 3.58 ± 0.29 μM and 23.94 ± 1.00 μM, respectively, compared to that of compound 27 (IC50 = 19.67 ± 1.12 μM). Importantly, both SW-b and SW-d exhibited low cytotoxicity, with CC50 values of 24.65 μmol and 133.70 μmol, respectively, resulting in selectivity indices of 6.89 and 5.58, respectively. Furthermore, when compared to the positive control compound 3′-dATP (IC50 = 30.09 ± 8.26 μM), SW-b and SW-d displayed superior inhibitory activity in an enzyme inhibitory assay, with IC50 values of 11.54 ± 1.30 μM and 13.54 ± 0.32 μM, respectively. Molecular dynamics (MD) simulations elucidated the mode of action of SW-b and SW-d, highlighting their ability to enhance π–π packing interactions between benzene rings and residue W795 in the S1 fragment, compared to compounds 27 and 29. Although the transacylsulphonamide fragment reduced the interaction between T794 and NH, it augmented the interaction between R729 and T794. In summary, our study underscores the potential of SW-b and SW-d as allosteric inhibitors targeting the DENV NS5 RdRp domain. However, further in vivo studies are warranted to assess their pharmacology and toxicity profiles.

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“…The replication of dengue virus in cell culture was reported as early as 1950 , and has since been performed in a large variety of mammalian and insect cells. Today, the preferred in vitro systems for the study of DENV infections are Huh-7 and Vero cells, and detailed antiviral screening protocols, amenable for high-throughput screening, are available. , Reporter gene and subgenomic replicon assays were also developed. Scheme provides an overview of cellular assay systems for biological studies and the characterization of antiviral agents.…”

Section: Transmission and Ecology Of Dengue Virusmentioning

confidence: 99%

The Medicinal Chemistry of Dengue Virus

et al. 2016

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The dengue virus and related flaviviruses are an increasing global health threat. In this perspective, we comment on and review medicinal chemistry efforts aimed at the prevention or treatment of dengue infections. We include target-based approaches aimed at viral or host factors and results from phenotypic screenings in cellular assay systems for viral replication. This perspective is limited to the discussion of results that provide explicit chemistry or structure-activity relationship (SAR), or appear to be of particular interest to the medicinal chemist for other reasons. The discovery and development efforts discussed here may at least partially be extrapolated toward other emerging flaviviral infections, such as West Nile virus. Therefore, this perspective, although not aimed at flaviviruses in general, should also be able to provide an overview of the medicinal chemistry of these closely related infectious agents.

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Cell-Based Antiviral Assays for Screening and Profiling Inhibitors Against Dengue Virus

Cited by 4 publications

References 3 publications

Blocking NS3–NS4B interaction inhibits dengue virus in non-human primates

Blocking NS3–NS4B interaction inhibits dengue virus in non-human primates

Design, Synthesis, Evaluation and Molecular Dynamics Simulation of Dengue Virus NS5-RdRp Inhibitors

The Medicinal Chemistry of Dengue Virus

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