JIB-04 Has Broad-Spectrum Antiviral Activity and Inhibits SARS-CoV-2 Replication and Coronavirus Pathogenesis

Pathogenic coronaviruses represent a major threat to global public health. Here, using a recombinant reporter virus-based compound screening approach, we identified several small-molecule inhibitors that potently block the replication of the newly emerged severe acute respiratory syndrome virus 2 (SARS-CoV-2). Two compounds, nitazoxanide and JIB-04 inhibited SARS-CoV-2 replication in Vero E6 cells with an EC 50 of 4.90 μM and 0.69 μM, respectively, with specificity indices of greater than 150. Both inhibitors had in vitro antiviral activity in multiple cell types against some DNA and RNA viruses, including porcine transmissible gastroenteritis virus. In an in vivo porcine model of coronavirus infection, administration of JIB-04 reduced virus infection and associated tissue pathology, which resulted in improved body weight gain and survival. These results highlight the potential utility of nitazoxanide and JIB-04 as antiviral agents against SARS-CoV-2 and other viral pathogens.

[1]  D. Fremont,et al.  Neutralizing Antibody and Soluble ACE2 Inhibition of a Replication-Competent VSV-SARS-CoV-2 and a Clinical Isolate of SARS-CoV-2 , 2020, Cell Host & Microbe.

[2]  S. Cherry,et al.  Drug repurposing screens reveal FDA approved drugs active against SARS-Cov-2 , 2020, bioRxiv.

[3]  John G. Doench,et al.  Genome-wide CRISPR screen reveals host genes that regulate SARS-CoV-2 infection , 2020, bioRxiv.

[4]  D. Fremont,et al.  Cholesterol 25-hydroxylase suppresses SARS-CoV-2 replication by blocking membrane fusion , 2020, Proceedings of the National Academy of Sciences.

[5]  D. Fremont,et al.  Neutralizing Antibody and Soluble ACE2 Inhibition of a Replication-Competent VSV-SARS-CoV-2 and a Clinical Isolate of SARS-CoV-2. , 2020, SSRN.

[6]  Matthew J. O’Meara,et al.  Morphological Cell Profiling of SARS-CoV-2 Infection Identifies Drug Repurposing Candidates for COVID-19 , 2020, bioRxiv.

[7]  Lisa E. Gralinski,et al.  SARS-CoV-2 Reverse Genetics Reveals a Variable Infection Gradient in the Respiratory Tract , 2020, Cell.

[8]  Wu Zhong,et al.  The anti-influenza virus drug, arbidol is an efficient inhibitor of SARS-CoV-2 in vitro , 2020, Cell Discovery.

[9]  M. Diamond,et al.  TMPRSS2 and TMPRSS4 mediate SARS-CoV-2 infection of human small intestinal enterocytes , 2020, bioRxiv.

[10]  Yolanda T. Chong,et al.  Identification of potential treatments for COVID-19 through artificial intelligence-enabled phenomic analysis of human cells infected with SARS-CoV-2 , 2020, bioRxiv.

[11]  Hualiang Jiang,et al.  Structure-based design of antiviral drug candidates targeting the SARS-CoV-2 main protease , 2020, Science.

[12]  Timothy K. Soh,et al.  Inhibition of PIKfyve kinase prevents infection by Zaire ebolavirus and SARS-CoV-2 , 2020, bioRxiv.

[13]  Peter G. Schultz,et al.  A Large-scale Drug Repositioning Survey for SARS-CoV-2 Antivirals , 2020, bioRxiv.

[14]  L. Guddat,et al.  Structure of the RNA-dependent RNA polymerase from COVID-19 virus , 2020, Science.

[15]  Yan Zhang,et al.  Structural Basis for the Inhibition of the RNA-Dependent RNA Polymerase from SARS-CoV-2 by Remdesivir , 2020, bioRxiv.

[16]  Shinji Makino,et al.  An Infectious cDNA Clone of SARS-CoV-2 , 2020, Cell Host & Microbe.

[17]  Hualiang Jiang,et al.  Structure of Mpro from SARS-CoV-2 and discovery of its inhibitors , 2020, Nature.

[18]  Xiaotao Lu,et al.  An orally bioavailable broad-spectrum antiviral inhibits SARS-CoV-2 in human airway epithelial cell cultures and multiple coronaviruses in mice , 2020, Science Translational Medicine.

[19]  Bruno Coutard,et al.  In vitro screening of a FDA approved chemical library reveals potential inhibitors of SARS-CoV-2 replication , 2020, Scientific Reports.

[20]  Hualiang Jiang,et al.  Structure-Based Design, Synthesis and Biological Evaluation of Peptidomimetic Aldehydes as a Novel Series of Antiviral Drug Candidates Targeting the SARS-CoV-2 Main Protease , 2020, bioRxiv.

[21]  Yan Liu,et al.  Characterization of spike glycoprotein of SARS-CoV-2 on virus entry and its immune cross-reactivity with SARS-CoV , 2020, Nature Communications.

[22]  R. Hilgenfeld,et al.  Crystal structure of SARS-CoV-2 main protease provides a basis for design of improved α-ketoamide inhibitors , 2020, Science.

[23]  K. He,et al.  Characterization and evaluation of the pathogenicity of a natural recombinant transmissible gastroenteritis virus in China , 2020, Virology.

[24]  G. Herrler,et al.  SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor , 2020, Cell.

[25]  Wu Zhong,et al.  Remdesivir and chloroquine effectively inhibit the recently emerged novel coronavirus (2019-nCoV) in vitro , 2020, Cell Research.

[26]  E. Holmes,et al.  A new coronavirus associated with human respiratory disease in China , 2020, Nature.

[27]  Kai Zhao,et al.  A pneumonia outbreak associated with a new coronavirus of probable bat origin , 2020, Nature.

[28]  S. Whelan,et al.  Inhibition of PIKfyve kinase prevents infection by EBOV and SARS-CoV-2 , 2020 .

[29]  S. Schultz-Cherry,et al.  Astrovirus Replication Is Inhibited by Nitazoxanide In Vitro and In Vivo , 2019, Journal of Virology.

[30]  Rong-li Guo,et al.  Aminopeptidase N-null neonatal piglets are protected from transmissible gastroenteritis virus but not porcine epidemic diarrhea virus , 2019, Scientific Reports.

[31]  A. Ward,et al.  Structure of the SARS-CoV nsp12 polymerase bound to nsp7 and nsp8 co-factors , 2019, Nature Communications.

[32]  R. Flavell,et al.  Rotavirus VP3 targets MAVS for degradation to inhibit type III interferon expression in intestinal epithelial cells , 2018, eLife.

[33]  M. Peppelenbosch,et al.  Nitazoxanide Inhibits Human Norovirus Replication and Synergizes with Ribavirin by Activation of Cellular Antiviral Response , 2018, Antimicrobial Agents and Chemotherapy.

[34]  J. Diedrich,et al.  SHMT2 and the BRCC36/BRISC deubiquitinase regulate HIV-1 Tat K63-ubiquitylation and destruction by autophagy , 2018, PLoS pathogens.

[35]  T. Shirao,et al.  Drebrin restricts rotavirus entry by inhibiting dynamin-mediated endocytosis , 2017, Proceedings of the National Academy of Sciences.

[36]  Tero Aittokallio,et al.  SynergyFinder: a web application for analyzing drug combination dose–response matrix data , 2017, Bioinform..

[37]  Steven H Kleinstein,et al.  Dynamic expression profiling of type I and type III interferon‐stimulated hepatocytes reveals a stable hierarchy of gene expression , 2014, Hepatology.

[38]  A. Iwasaki,et al.  Epigenetic Reprogramming of the Type III Interferon Response Potentiates Antiviral Activity and Suppresses Tumor Growth , 2014, PLoS biology.

[39]  A. Ciucci,et al.  Thiazolides, a New Class of Antiviral Agents Effective against Rotavirus Infection, Target Viral Morphogenesis, Inhibiting Viroplasm Formation , 2013, Journal of Virology.

[40]  S. Peña-Llopis,et al.  A small molecule modulates Jumonji histone demethylase activity and selectively inhibits cancer growth , 2013, Nature Communications.

[41]  Chang-Seon Song,et al.  Nitazoxanide suppresses IL-6 production in LPS-stimulated mouse macrophages and TG-injected mice. , 2012, International immunopharmacology.

[42]  Karen K. Y. Lam,et al.  Nitazoxanide Stimulates Autophagy and Inhibits mTORC1 Signaling and Intracellular Proliferation of Mycobacterium tuberculosis , 2012, PLoS pathogens.

[43]  J. Puglisi,et al.  The anti-hepatitis C agent nitazoxanide induces phosphorylation of eukaryotic initiation factor 2alpha via protein kinase activated by double-stranded RNA activation. , 2009, Gastroenterology.

[44]  L. Saravolatz,et al.  Nitazoxanide: a new thiazolide antiparasitic agent. , 2005, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[45]  N. Perrimon,et al.  Genome-wide RNAi screen reveals a specific sensitivity of IRES-containing RNA viruses to host translation inhibition. , 2005, Genes & development.

[46]  L. Saif.  Animal coronavirus vaccines: lessons for SARS. , 2004, Developments in biologicals.

[47]  M. E. Hahn,et al.  Regulatory Interactions among Three Members of the Vertebrate Aryl Hydrocarbon Receptor Family: AHR Repressor, AHR1, and AHR2* , 2002, The Journal of Biological Chemistry.

[48]  F. Belpaire,et al.  Selective serotonin reuptake inhibitors and cytochrome P-450 mediated drug-drug interactions: an update. , 2002, Current drug metabolism.

[49]  H Maisonneuve,et al.  Nitazoxanide in the treatment of Taenia saginata and Hymenolepis nana infections. , 1984, The American journal of tropical medicine and hygiene.