The molecular tweezer CLR01 inhibits Ebola and Zika virus infection

Röcker, Annika; Müller, Janis A.; Dietzel, Erik; Harms, Mirja; Krüger, Franziska; Heid, Christian; Sowislok, Andrea; Riber, Camilla Frich; Kupke, Alexandra; Lippold, Sina; Einem, Jens von; Beer, Judith; Knöll, Bernd; Becker, Stephan; Schmidt‐Chanasit, Jonas; Otto, Markus; Vapalahti, Olli; Zelikin, Alexander N.; Bitan, Gal; Schräder, Thomas; Münch, Jan

doi:10.1016/j.antiviral.2018.02.003

Cited by 33 publications

(51 citation statements)

References 53 publications

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“…Inspired by the well-established Zika virus infection assay that quantifies the viral envelope (E) protein by a horseradish peroxidase (HRP) coupled antibody ( Aubry et al, 2016 ; Conzelmann et al, 2019 ; Müller et al, 2018 , 2017 ; Röcker et al, 2018 ) we here aimed to detect the spike (S) protein of SARS-CoV-2 by a similar approach. To adapt this in-cell ELISA to measure SARS-CoV-2 infection, we made use of the anti-SARS-CoV/SARS-CoV-2 antibody 1A9 that targets the highly conserved loop region between the HR1 and HR2 in the S2 subunit of the S protein ( Ng et al, 2014 ; Walls et al, 2020 ) ( www.nextstrain.org ( Hadfield et al, 2018 )) and has been verified to bind to SARS-CoV-2 S protein ( Zheng et al, 2020 ).…”

Section: Resultsmentioning

confidence: 99%

An enzyme-based immunodetection assay to quantify SARS-CoV-2 infection

et al. 2020

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SARS-CoV-2 is a novel pandemic coronavirus that caused a global health and economic crisis. The development of efficient drugs and vaccines against COVID-19 requires detailed knowledge about SARS-CoV-2 biology. Several techniques to detect SARS-CoV-2 infection have been established, mainly based on counting infected cells by staining plaques or foci, or by quantifying the viral genome by PCR. These methods are laborious, time-consuming and expensive and therefore not suitable for a high sample throughput or rapid diagnostics. We here report a novel enzyme-based immunodetection assay that directly quantifies the amount of de novo synthesized viral spike protein within fixed and permeabilized cells. This in-cell ELISA enables a rapid and quantitative detection of SARS-CoV-2 infection in microtiter format, regardless of the virus isolate or target cell culture. It follows the established method of performing ELISA assays and does not require expensive instrumentation. Utilization of the in-cell ELISA allows to e.g. determine TCID 50 of virus stocks, antiviral efficiencies (IC 50 values) of drugs or neutralizing activity of sera. Thus, the in-cell spike ELISA represents a promising alternative to study SARS-CoV-2 infection and inhibition and may facilitate future research.

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Section: Resultsmentioning

confidence: 99%

An enzyme-based immunodetection assay to quantify SARS-CoV-2 infection

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“…The cholesterol dependence of DENV entry and post-entry steps have been observed in several mammalian cell lines (Lee et al, 2008 ; Soto-Acosta et al, 2013 ; Martínez-Gutierrez et al, 2014 ), it does not seem to be a general event, because it has been described that cholesterol is not required for Vero (green monkey epithelial kidney cell line) (Acosta et al, 2009 ; Carro and Damonte, 2013 ), HepG2 (Hepatocarcinoma cell line) and ECV304 cells entry (human endothelial cell line) (Rothwell et al, 2009 ). In addition, it has been described that the molecular tweezer CLR01, a small molecule that previously has been shown to inactivate some viruses through a selective interaction with the host-membrane-derived lipid bilayer of the viral envelope, inhibit EBOV (ebola virus) and ZIKV infection (Röcker et al, 2018 ).…”

Section: The Role Of Host Cholesterol During the Flavivirus Life Cyclmentioning

confidence: 99%

The Role of Host Cholesterol During Flavivirus Infection

Osuna-Ramos

Reyes-Ruiz

Ángel

2018

Front. Cell. Infect. Microbiol.

132

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In recent years the emergence and resurgence of arboviruses have generated a global health alert. Among arboviruses, Dengue (DENV), Zika (ZIKV), Yellow Fever (YFV), and West Nile (WNV) virus, belong to the genus Flavivirus, cause high viremia and occasionally fatal clinical disease in humans. Given the genetic austerity of the virus, they depend on cellular factors and organelles to complete its replication. One of the cellular components required for flavivirus infection is cholesterol. Cholesterol is an abundant lipid in biomembranes of eukaryotes cells and is necessary to maintain the cellular homeostasis. Recently, it has been reported, that cholesterol is fundamental during flavivirus infection in both mammal and insect vector models. During infection with DENV, ZIKV, YFV, and WNV the modulation of levels of host-cholesterol facilitates viral entry, replicative complexes formation, assembly, egress, and control of the interferon type I response. This modulation involves changes in cholesterol uptake with the concomitant regulation of cholesterol receptors as well as changes in cholesterol synthesis related to important modifications in cellular metabolism pathways. In view of the flavivirus dependence of cholesterol and the lack of an effective anti-flaviviral treatment, this cellular lipid has been proposed as a therapeutic target to treat infection using FDA-approved cholesterol-lowering drugs. This review aims to address the dependence of cholesterol by flaviviruses as well as the basis for anti flaviviral therapy using drugs which target is cholesterol synthesis or uptake.

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“… 15 , 17 , 22 , 23 Unexpectedly, we previously found that CLR01 not only antagonizes the infectivity-enhancing activity of seminal amyloids but also exerts a direct antiviral activity against several enveloped viruses such as HIV-1, hepatitis c virus (HCV), herpes simplex virus type 2 (HSV-2), human cytomegalovirus (HCMV), Zika virus (ZIKV), and Ebola virus. 14 , 24 The antiviral activity is a consequence of the direct interaction of CLR01 with the membrane of the enveloped viral particle, which ultimately results in the loss of virion integrity and hence infectivity. 14 , 24 However, the precise antiviral mechanism of CLR01 remains unclear.…”

Section: Introductionmentioning

confidence: 99%

Supramolecular Mechanism of Viral Envelope Disruption by Molecular Tweezers

et al. 2020

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Broad-spectrum antivirals are powerful weapons against dangerous viruses where no specific therapy exists, as in the case of the ongoing SARS-CoV-2 pandemic. We discovered that a lysine- and arginine-specific supramolecular ligand (CLR01) destroys enveloped viruses, including HIV, Ebola, and Zika virus, and remodels amyloid fibrils in semen that promote viral infection. Yet, it is unknown how CLR01 exerts these two distinct therapeutic activities. Here, we delineate a novel mechanism of antiviral activity by studying the activity of tweezer variants: the “phosphate tweezer” CLR01, a “carboxylate tweezer” CLR05, and a “phosphate clip” PC. Lysine complexation inside the tweezer cavity is needed to antagonize amyloidogenesis and is only achieved by CLR01. Importantly, CLR01 and CLR05 but not PC form closed inclusion complexes with lipid head groups of viral membranes, thereby altering lipid orientation and increasing surface tension. This process disrupts viral envelopes and diminishes infectivity but leaves cellular membranes intact. Consequently, CLR01 and CLR05 display broad antiviral activity against all enveloped viruses tested, including herpesviruses, Measles virus, influenza, and SARS-CoV-2. Based on our mechanistic insights, we potentiated the antiviral, membrane-disrupting activity of CLR01 by introducing aliphatic ester arms into each phosphate group to act as lipid anchors that promote membrane targeting. The most potent ester modifications harbored unbranched C4 units, which engendered tweezers that were approximately one order of magnitude more effective than CLR01 and nontoxic. Thus, we establish the mechanistic basis of viral envelope disruption by specific tweezers and establish a new class of potential broad-spectrum antivirals with enhanced activity.

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The molecular tweezer CLR01 inhibits Ebola and Zika virus infection

Cited by 33 publications

References 53 publications

An enzyme-based immunodetection assay to quantify SARS-CoV-2 infection

An enzyme-based immunodetection assay to quantify SARS-CoV-2 infection

The Role of Host Cholesterol During Flavivirus Infection

Supramolecular Mechanism of Viral Envelope Disruption by Molecular Tweezers

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