GNS561 Exhibits Potent Antiviral Activity against SARS-CoV-2 through Autophagy Inhibition

Since December 2019, SARS-CoV-2 has spread quickly worldwide, leading to more than 280 million confirmed cases, including over 5,000,000 deaths. Interestingly, coronaviruses were found to subvert and hijack autophagic process to allow their viral replication. Autophagy-modulating compounds thus rapidly emerged as an attractive strategy to fight SARS-CoV-2 infection, including the well-known chloroquine (CQ). Here, we investigated the antiviral activity and associated mechanism of GNS561/Ezurpimtrostat, a small lysosomotropic molecule inhibitor of late-stage autophagy. Interestingly, GNS561 exhibited antiviral activity of 6–40 nM depending on the viral strain considered, currently positioning it as the most powerful molecule investigated in SARS-CoV-2 infection. We then showed that GNS561 was located in lysosome-associated-membrane-protein-2-positive (LAMP2-positive) lysosomes, together with SARS-CoV-2. Moreover, GNS561 increased LC3-II spot size and caused the accumulation of autophagic vacuoles and the presence of multilamellar bodies, suggesting that GNS561 disrupted the autophagy mechanism. To confirm our findings, we used the K18-hACE2 mouse model and highlighted that GNS561 treatment led to a decline in SARS-CoV-2 virions in the lungs associated with a disruption of the autophagy pathway. Overall, our study highlights GNS561 as a powerful drug in the treatment of SARS-CoV-2 infection and supports the hypothesis that autophagy blockers could be an alternative strategy for COVID-19.

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GNS561 exhibits potent in vitro antiviral activity against SARS-CoV-2 through autophagy inhibition

10.1101/2020.10.06.327635 ◽

2020 ◽

Author(s):

Philippe Halfon ◽

Eloïne Bestion ◽

Keivan Zandi ◽

Julien Andreani ◽

Jean-Pierre Baudoin ◽

...

Keyword(s):

Severe Acute Respiratory Syndrome ◽

Antiviral Activity ◽

Antiviral Effect ◽

Therapeutic Strategies ◽

Multilamellar Bodies ◽

Molecule Inhibitor ◽

Lipophilic Molecule ◽

Autophagic Process ◽

Autophagy Inhibition

AbstractSince December 2019, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2/2019-nCoV) has spread quickly worldwide, with more than 29 million cases and 920,000 deaths. Interestingly, coronaviruses were found to subvert and hijack the autophagic process to allow their viral replication. One of the spotlights had been focused on the autophagy inhibitors as a target mechanism effective in the inhibition of SARS-CoV-2 infection. Consequently, chloroquine (CQ) and hydroxychloroquine (HCQ), a derivative of CQ, was suggested as the first potentially be therapeutic strategies as they are known to be autophagy inhibitors. Then, they were used as therapeutics in SARS-CoV-2 infection along with remdesivir, for which the FDA approved emergency use authorization. Here, we investigated the antiviral activity and associated mechanism of GNS561, a small basic lipophilic molecule inhibitor of late-stage autophagy, against SARS-CoV-2. Our data indicated that GNS561 showed the highest antiviral effect for two SARS-CoV-2 strains compared to CQ and remdesivir. Focusing on the autophagy mechanism, we showed that GNS561, located in LAMP2-positive lysosomes, together with SARS-CoV-2, blocked autophagy by increasing the size of LC3-II spots and the accumulation of autophagic vacuoles in the cytoplasm with the presence of multilamellar bodies characteristic of a complexed autophagy. Finally, our study revealed that the combination of GNS561 and remdesivir was associated with a strong synergistic antiviral effect against SARS-CoV-2. Overall, our study highlights GNS561 as a powerful drug in SARS-CoV-2 infection and supports that the hypothesis that autophagy inhibitors could be an alternative strategy for SARS-CoV-2 infection.

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Antiviral Activity of (−)-Carbocyclic Cytosine [(−)-Carbodine] Against Venezuelan Equine Encephalitis Virus (VEEV) in a Mouse Model

Antiviral Research ◽

10.1016/j.antiviral.2007.01.050 ◽

2007 ◽

Vol 74 (3) ◽

pp. A44-A45

Author(s):

J JULANDER ◽

C CHU ◽

J RAO ◽

K SHAFER ◽

J MORREY

Keyword(s):

Antiviral Activity ◽

Mouse Model ◽

Venezuelan Equine Encephalitis Virus ◽

Encephalitis Virus ◽

Venezuelan Equine Encephalitis ◽

Equine Encephalitis Virus

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Outer Membrane Protein F (Porin) Preparation of Pseudomonas aeruginosa as a Protective Vaccine Against Heterologous Immunotype Strains in a Burned Mouse Model

The Journal of Infectious Diseases ◽

10.1093/infdis/155.6.1282 ◽

1987 ◽

Vol 155 (6) ◽

pp. 1282-1291 ◽

Cited By ~ 55

Author(s):

J. M. Matthews-Greer ◽

H. E. Gilleland

Keyword(s):

Pseudomonas Aeruginosa ◽

Membrane Protein ◽

Mouse Model ◽

Outer Membrane ◽

Outer Membrane Protein ◽

Protein F ◽

Outer Membrane Protein F

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Bivalent Inhibitor with Selectivity for Trimeric MMP-9 Amplifies Neutrophil Chemotaxis and Enables Functional Studies on MMP-9 Proteoforms

Cells ◽

10.3390/cells9071634 ◽

2020 ◽

Vol 9 (7) ◽

pp. 1634

Author(s):

Elisa Nuti ◽

Armando Rossello ◽

Doretta Cuffaro ◽

Caterina Camodeca ◽

Jens Van Bael ◽

...

Keyword(s):

Mouse Model ◽

Inflammatory Cell ◽

High Selectivity ◽

Neutrophil Migration ◽

Endotoxin Shock ◽

Flow Cytometry Analysis ◽

Functional Studies ◽

Fundamental Part ◽

Molecule Inhibitor ◽

Interesting Approach

A fundamental part of the immune response to infection or injury is leukocyte migration. Matrix metalloproteinases (MMPs) are a class of secreted or cell-bound endopeptidases, implicated in every step of the process of inflammatory cell migration. Hence, specific inhibition of MMPs is an interesting approach to control inflammation. We evaluated the potential of a bivalent carboxylate inhibitor to selectively inhibit the trimeric proteoform of MMP-9 and compared this with a corresponding monovalent inhibitor. The bivalent inhibitor efficiently inhibited trimeric MMP-9 (IC50 = 0.1 nM), with at least 500-fold selectivity for MMP-9 trimers over monomers. Surprisingly, in a mouse model for chemotaxis, the bivalent inhibitor amplified leukocyte influxes towards lipopolysaccharide-induced inflammation. We verified by microscopic and flow cytometry analysis increased amounts of neutrophils. In a mouse model for endotoxin shock, mice treated with the bivalent inhibitor had significantly increased levels of MMP-9 in plasma and lungs, indicative for increased inflammation. In conclusion, we propose a new role for MMP-9 trimers in tempering excessive neutrophil migration. In addition, we have identified a small molecule inhibitor with a high selectivity for the trimeric proteoform of MMP-9, which will allow further research on the functions of MMP-9 proteoforms.

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Protective immunity against Rickettsia heilongjiangensis in a C3H/HeN mouse model mediated by outer membrane protein B-pulsed dendritic cells

Science China Life Sciences ◽

10.1007/s11427-014-4720-4 ◽

2014 ◽

Vol 58 (3) ◽

pp. 287-296 ◽

Cited By ~ 3

Author(s):

YanFen Meng ◽

XiaoLu Xiong ◽

Yong Qi ◽

ChangSong Duan ◽

WenPing Gong ◽

...

Keyword(s):

Dendritic Cells ◽

Membrane Protein ◽

Mouse Model ◽

Outer Membrane ◽

Outer Membrane Protein ◽

Protective Immunity ◽

Rickettsia Heilongjiangensis ◽

Protein B

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Enhancement of Pulmonary Clearance ofMoraxella (Branhamella) catarrhalisfollowing Immunization with Outer Membrane Protein CD in a Mouse Model

The Journal of Infectious Diseases ◽

10.1086/314501 ◽

1998 ◽

Vol 178 (6) ◽

pp. 1667-1675 ◽

Cited By ~ 46

Author(s):

Timothy F. Murphy ◽

Jennelle M. Kyd ◽

Anna John ◽

Charmaine Kirkham ◽

Allan W. Cripps

Keyword(s):

Membrane Protein ◽

Mouse Model ◽

Outer Membrane ◽

Outer Membrane Protein ◽

Pulmonary Clearance

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A small molecule inhibitor of MyD88 exhibits broad spectrum antiviral activity by up regulation of type I interferon

Antiviral Research ◽

10.1016/j.antiviral.2020.104854 ◽

2020 ◽

Vol 181 ◽

pp. 104854

Author(s):

Kamal U. Saikh ◽

Elaine M. Morazzani ◽

Ashley E. Piper ◽

Russell R. Bakken ◽

Pamela J. Glass

Keyword(s):

Antiviral Activity ◽

Small Molecule ◽

Broad Spectrum ◽

Type I Interferon ◽

Small Molecule Inhibitor ◽

Type I ◽

Molecule Inhibitor

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Outer Membrane Protein A (OmpA) of Shigella flexneri 2a, Induces Protective Immune Response in a Mouse Model

PLoS ONE ◽

10.1371/journal.pone.0022663 ◽

2011 ◽

Vol 6 (7) ◽

pp. e22663 ◽

Cited By ~ 56

Author(s):

Debasis Pore ◽

Nibedita Mahata ◽

Amit Pal ◽

Manoj K. Chakrabarti

Keyword(s):

Immune Response ◽

Membrane Protein ◽

Mouse Model ◽

Outer Membrane ◽

Outer Membrane Protein ◽

Shigella Flexneri ◽

Protein A ◽

Protective Immune Response ◽

Shigella Flexneri 2A ◽

Outer Membrane Protein A

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A Novel Substance Purified from Perilla Frutescens Britton Inhibits an Early Stage of HIV-1 Replication without Blocking Viral Adsorption

Antiviral Chemistry and Chemotherapy ◽

10.1177/095632020201300503 ◽

2002 ◽

Vol 13 (5) ◽

pp. 283-288 ◽

Cited By ~ 13

Author(s):

T Kawahata ◽

T Otake ◽

H Mori ◽

Y Kojima ◽

I Oishi ◽

...

Keyword(s):

Antiviral Activity ◽

Viral Entry ◽

Cytopathic Effect ◽

Early Stage ◽

Perilla Frutescens ◽

Viral Strain ◽

Viral Particles ◽

Viral Adsorption ◽

Hiv 1

Pf-gp6, a 6 kDa anti-degranulation glycoprotein purified from the extract of Perilla frutescens, was examined for its antiviral activity against HIV-1 and HIV-2 in vitro. HIV-1-induced cytopathic effect and proviral DNA synthesis were inhibited in the presence of Pf-gp6. The 50% inhibitory concentrations of Pf-gp6 for various HIV-1 strains, including clinical isolates and CCR5-using (R5) HIV-1, ranged between 1.3 and 71.0 μg/ml, depending on the combination of viral strain and host cell. Furthermore, Pf-gp6 did not directly inactivate infectious viral particles. A time-of-addition experiment revealed that Pf-gp6 lost its activity before zidovudine but after the CXCR-4 antagonist AMD3100 during the early stage of viral infection. Although the pinpoint target of Pf-gp6 remains to be elucidated, it may interfere with a step between viral entry and reverse transcription.

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