Identification of a Small-Molecule Entry Inhibitor for Filoviruses

Basu, Arnab; Li, Bing; Mills, Debra M.; Panchal, Rekha G.; Cardinale, Steven C.; Butler, Michelle M.; Peet, Norton P.; Majgier‐Baranowska, Helena; Williams, John D.; Patel, Ishan; Moir, Donald T.; Bavari, Sina; Ray, Ranjit; Farzan, Michael; Rong, Lijun; Bowlin, Terry L.

doi:10.1128/jvi.01456-10

Cited by 101 publications

(103 citation statements)

References 63 publications

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“…In contrast to previous reports of anti-EBOV inhibitors that target either EBOV glycoprotein (GP) or viral polymerase by a limited number of chemical scaffolds or entities (3,5,14,25), the GPCR antagonists that showed antifiloviral entry activity here represent structurally different scaffolds and target different GPCRs, including histamine receptor, 5-HT (serotonin) receptor, muscarinic acetylcholine receptor, and adrenergic receptor (Fig. 1).…”

Section: Discussionmentioning

confidence: 63%

“…GPCR antagonists block filoviral entry at a postattachment step. To investigate if GPCR antagonists blocked the early attachment/binding step or the postbinding steps (e.g., fusion) of MARV, a "time-of-addition experiment" (14) was performed on MARV/HIV pseudovirions with two GPCR antagonists, benztropine mesylate and cyproheptadine (a promiscuous GPCR antagonist), using heparin, bafilomycin A1, and zidovudine (AZT) as controls. As expected, heparin, which was shown by others and us to block the early attachment/binding of filoviruses to the target cells (6, 7), exerted its inhibitory effect at early time points (Ϫ1 and 0 h), while bafilomycin A1 (an inhibitor of vacuolar-type H ϩ -ATPase (15) and AZT (an HIV reverse transcriptase inhibitor [16]) were still effective at the late time points (Ͼ6 h).…”

Section: Gpcr Antagonists Inhibit the Ebov/marv Gp-mediated Viral Entrymentioning

confidence: 99%

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Inhibition of Ebola and Marburg Virus Entry by G Protein-Coupled Receptor Antagonists

Cheng

Lear-Rooney

Johansen

et al. 2015

J Virol

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107

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Filoviruses, consisting of Ebola virus (EBOV) and Marburg virus (MARV), are among the most lethal infectious threats to mankind. Infections by these viruses can cause severe hemorrhagic fevers in humans and nonhuman primates with high mortality rates. Since there is currently no vaccine or antiviral therapy approved for humans, there is an urgent need to develop prophylactic and therapeutic options for use during filoviral outbreaks and bioterrorist attacks. One of the ideal targets against filoviral infection and diseases is at the entry step, which is mediated by the filoviral glycoprotein (GP). In this report, we screened a chemical library of small molecules and identified numerous inhibitors, which are known G protein-coupled receptor (GPCR) antagonists targeting different GPCRs, including histamine receptors, 5-HT (serotonin) receptors, muscarinic acetylcholine receptor, and adrenergic receptor. These inhibitors can effectively block replication of both infectious EBOV and MARV, indicating a broad antiviral activity of the GPCR antagonists. The time-of-addition experiment and microscopic studies suggest that GPCR antagonists block filoviral entry at a step following the initial attachment but prior to viral/cell membrane fusion. These results strongly suggest that GPCRs play a critical role in filoviral entry and GPCR antagonists can be developed as an effective anti-EBOV/MARV therapy. IMPORTANCE Infection of Ebola virus and Marburg virus can cause severe illness in humans witha high mortality rate, and currently there is no FDA-approved vaccine or therapeutic treatment available. The 2013-2015 epidemic in West Africa underscores a lack of our understanding in the infection and pathogenesis of these viruses and the urgency of drug discovery and development. In this study, we have identified numerous inhibitors that are known G protein-coupled receptor (GPCR) antagonists targeting different GPCRs. These inhibitors can effectively block replication of both infectious EBOV and MARV, indicating a broad antiviral activity of the GPCR antagonists. Our results strongly suggest that GPCRs play a critical role in filoviral entry and GPCR antagonists can be developed as an effective anti-EBOV/MARV therapy.

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Section: Discussionmentioning

confidence: 63%

Section: Gpcr Antagonists Inhibit the Ebov/marv Gp-mediated Viral Entrymentioning

confidence: 99%

Inhibition of Ebola and Marburg Virus Entry by G Protein-Coupled Receptor Antagonists

Cheng

Lear-Rooney

Johansen

et al. 2015

J Virol

Self Cite

107

View full text Add to dashboard Cite

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“…As the viral envelope was the only difference between EBOVpp and the VSV and WSN pseudoviruses 19 , we determined whether EBOV GP was the potential target of the triterpenoid lead Y11 by a series of time-of-addition experiments 11,13,19 , including pretreatment of the viruses, pretreatment of the cells, and pre-attachment, postattachment and post-entry assays conducted in parallel, as previously reported (Supplementary Fig. 2a).…”

Section: Identification Of Ebov Gp As the Target By Time-of-addition mentioning

confidence: 99%

“…The "time-of-addition" experiment was designed as previously described 11,13,19 to determine the mechanism of action of the antiviral compounds, and the procedure is shown schematically in Supplementary Fig. 2.…”

Section: Time-of-addition Experimentmentioning

confidence: 99%

Triterpenoids Manipulate a Broad Range of Virus-Host Fusion via Wrapping the HR2 Domain Prevalent in Viral Envelopes

Si¹,

Meng²,

Tian³

et al. 2018

Preprint

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Recent years have witnessed a breakthrough in identification of a trimer-ofhairpins motif within viral envelopes that triggers a broad range of virus-host fusion.Identifying a domain capable of controlling virus-host fusion remains a challenge due to sequence diversity, heavy glycan shielding and multiple conformations. Here, we report that HR2, a prevalent heptad repeat sequence comprising an alpha-helical coil anchored in viral membranes, is an accessible site to triterpenes, a class of widely distributed natural products. Triterpenes and their derivatives inhibit the entry of Ebola, HIV, and influenza A viruses with distinct structure-activity relationships. Specifically, triterpenoid probes, upon activation by ultraviolet light, capture the viral envelope via crosslinking the HR2 coil. Profiling the Ebola HR2 sequence using amino acid substitution, surface plasmon resonance (SPR) and nuclear magnetic resonance (NMR) spectroscopy disclosed six constitutive residues that are accessible to triterpenoids, leading to wrapping of the hydrophobic helix by triterpenoids and blocking of the HR1-HR2 interaction, which is critical in the trimer-of-hairpins formation. This finding was also observed in the envelopes of HIV and influenza A viruses and might potentially extend to a broader variety of viruses. Our findings might translate into a shared mechanism that host utilize natural product triterpenoids to antagonize membrane fusion of respective viruses, complementing the current repertoire of antiviral agents.

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“…In previous MARV studies, the lentiviral NL4-3.Luc.R-E-was the major pseudovirus system used. [10][11][12][13] SG3 is another widely used HIV based pseudovirus system. [16][17][18] However, the requirement of special cells containing reporter genes greatly limits its application.…”

Section: Discussionmentioning

confidence: 99%

A bioluminescent imaging mouse model for Marburg virus based on a pseudovirus system

Zhang

Liu

et al. 2017

Human Vaccines & Immunotherapeutics

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Marburg virus (MARV) can cause lethal hemorrhagic fever in humans. Handling of MARV is restricted to high-containment biosafety level 4 (BSL-4) facilities, which greatly impedes research into this virus. In this study, a high titer of MARV pseudovirus was generated through optimization of the HIV backbone vectors, the ratio of backbone vector to MARV glycoprotein expression vector, and the transfection reagents. An in vitro neutralization assay and an in vivo bioluminescent imaging mouse model for MARV were developed based on the pseudovirus. Protective serum against MARV was successfully induced in guinea pigs, which showed high neutralization activity in vitro and could also protect Balb/c mice from MARV pseudovirus infection in vivo. This system could be a convenient tool to enable the evaluation of vaccines and therapeutic drugs against MARV in non-BSL-4 laboratories.

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Identification of a Small-Molecule Entry Inhibitor for Filoviruses

Cited by 101 publications

References 63 publications

Inhibition of Ebola and Marburg Virus Entry by G Protein-Coupled Receptor Antagonists

Inhibition of Ebola and Marburg Virus Entry by G Protein-Coupled Receptor Antagonists

Triterpenoids Manipulate a Broad Range of Virus-Host Fusion via Wrapping the HR2 Domain Prevalent in Viral Envelopes

A bioluminescent imaging mouse model for Marburg virus based on a pseudovirus system

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