#Oseltamivir #resistance in #severe #influenza A #H1N1pdm09 #pneumonia and #ARDS: a #French multicenter observational cohort study (Clin Infect Dis., abstract)

[Source: Clinical Infectious Diseases Journal, full page: (LINK). Abstract, edited.]

Clin Infect Dis. 2019 Sep 20. pii: ciz904. doi: 10.1093/cid/ciz904. [Epub ahead of print]

Oseltamivir resistance in severe influenza A(H1N1)pdm09 pneumonia and acute respiratory distress syndrome: a French multicenter observational cohort study.

Behillil S1, May F2,3, Fourati S4, Luyt CE5, Chicheportiche T5, Sonneville R6, Tandjaoui-Lambiotte Y7, Roux D8, Guérin L9, Mayaux J10, Maury E11, Ferré A12, Georger JF13, Voiriot G14, Enouf V1, van der Werf S1, Dessap AM2,3, de Prost N2,3.

Author information: 1 Unité de Génétique Moléculaire des Virus à ARN et Centre National de Référence des Virus des Infections Respiratoires (dont la grippe), Institut Pasteur, Université Paris Diderot, Sorbonne Paris Cité, Paris, France. 2 Service de Réanimation Médicale, Hôpitaux Universitaires Henri Mondor-Albert Chenevier, Assistance Publique-Hôpitaux de Paris, Créteil, France. 3 Groupe de Recherche Clinique CARMAS, Université Paris-Est Créteil, IMRB, Créteil,  France. 4 Département de Microbiologie, Hôpitaux Universitaires Henri Mondor-Albert Chenevier, Assistance Publique-Hôpitaux de Paris, Créteil, France. 5 Service de Médecine Intensive Réanimation, Hôpital de La Pitié Salpétrière, Assistance Publique-Hôpitaux de Paris, Paris, France. 6 Service de Médecine Intensive Réanimation, Hôpital Bichat Claude Bernard, Assistance Publique-Hôpitaux de Paris, Paris, France. 7 Service de Réanimation médico-chirurgicale, Hôpital Avicenne, Assistance Publique-Hôpitaux de Paris, Bobigny, France. 8 Service de réanimation médico-chirurgicale, Hôpital Louis Mourier, Assistance Publique-Hôpitaux de Paris, Colombes,  France; IAME, Université Paris Diderot, Paris, France. 9 Service de réanimation médicale, Hôpital Bicètre, Assistance Publique-Hôpitaux de Paris, Le Kremlin-Bicètre, France. 10 Service de Réanimation Médicale et Pneumologie, Hôpital de La Pitié Salpétrière, Assistance Publique-Hôpitaux de Paris, Paris, France. 11 Service de Réanimation Médicale, Hôpital Saint-Antoine, Assistance Publique-Hôpitaux de Paris, Paris, France. 12 Service de Réanimation, Centre hospitalier de Versailles, Le Chesnay, France. 13 Service de Réanimation, Centre hospitalier Intercommunal de Villeneuve Saint-Georges, Villeneuve Saint-Georges, France. 14 Service de Réanimation Médicale, Hôpital Tenon, Assistance Publique-Hôpitaux de Paris, Paris, France.

 

Abstract

In a multicenter cohort study including 22 oseltamivir-treated patients with influenza A(H1N1)pdm09 acute respiratory distress syndrome, prevalence of the H275Y substitution in the neuraminidase, responsible for highly reduced sensitivity to oseltamivir, was 23%. Patients infected with the H275Y mutant virus had higher day-28 mortality than others (80% vs 12%; p=0.011).

© The Author(s) 2019. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.

KEYWORDS: Influenza A Virus, H1N1 Subtype; Oseltamivir; Pneumonia, Viral; Respiratory Distress Syndrome, Adult

PMID: 31538643 DOI: 10.1093/cid/ciz904

Keywords: Seasonal Influenza; H1N1pdm09; Antivirals; Drugs Resistance; Oseltamivir; Pneumonia; ARDS; France.

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Replicative #fitness of seasonal #influenza A viruses with decreased susceptibility to #baloxavir (J Infect Dis., abstract)

[Source: Journal of Infectious Diseases, full page: (LINK). Abstract, edited.]

Replicative fitness of seasonal influenza A viruses with decreased susceptibility to baloxavir

Anton Chesnokov, Mira C Patel, Vasiliy P Mishin, Juan A De La Cruz, Lori Lollis, Ha T Nguyen, Vivien Dugan, David E Wentworth, Larisa V Gubareva

The Journal of Infectious Diseases, jiz472, https://doi.org/10.1093/infdis/jiz472

Published: 21 September 2019

 

Abstract

Susceptibility of influenza A viruses to baloxavir can be affected by changes at amino acid residue 38 in polymerase acidic (PA) protein. Information on replicative fitness of PA-I38-substituted viruses remains sparse. We demonstrated that substitutions I38L/M/S/T not only had a differential effect on baloxavir susceptibility (9- to 116-fold), but also on in vitro replicative fitness. While I38L conferred undiminished growth, other substitutions led to mild attenuation. In a ferret model, control viruses outcompeted those carrying I38M or I38T substitutions, although their advantage was limited. These findings offer insights into the attributes of baloxavir resistant viruses needed for informed risk assessment.

Cap-dependent endonuclease inhibitor, replicative fitness, polymerase acidic protein, influenza, drug resistance, ferret, baloxavir acid, antiviral

Issue Section: Brief Report

This content is only available as a PDF.

Author notes

These authors contributed equally to this article and share first authorship

Published by Oxford University Press for the Infectious Diseases Society of America 2019. This work is written by (a) US Government employee(s) and is in the public domain in the US.

Keywords: Influenza A; Antivirals; Drugs Resistance; Baloxavir.

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#Azithromycin Protects against #Zika virus #Infection by Upregulating virus-induced Type I and III #Interferon Responses (Antimicrob Agents Chemother., abstract)

[Source: Antimicrobial Agents and Chemotherapy, full page: (LINK). Abstract, edited.]

Azithromycin Protects against Zika virus Infection by Upregulating virus-induced Type I and III Interferon Responses

Chunfeng Li, Shulong Zu, Yong-Qiang Deng, Dapei Li, Kislay Parvatiyar, Natalie Quanquin, Jingzhe Shang, Nina Sun, Jiaqi Su, Zhenyang Liu, Min Wang, Saba R. Aliyari, Xiao-Feng Li, Aiping Wu, Feng Ma, Yi Shi, Karin Nielsevn-Saines, Jae U. Jung, Frank Xiao-Feng Qin, Cheng-Feng Qin, Genhong Cheng

DOI: 10.1128/AAC.00394-19

 

ABSTRACT

Azithromycin (AZM) is a widely used antibiotic, with additional antiviral and anti-inflammatory properties that remain poorly understood. Although Zika virus (ZIKV) poses a significant threat to global health, there are currently no vaccines or effective therapeutics against it. Herein, we report that AZM effectively suppresses ZIKV infection in vitro by targeting a late stage in the viral life cycle. Besides that, AZM upregulates the expression of host type I and III interferons and several of their downstream interferon-stimulated genes (ISGs) in response to ZIKV infection. In particular, we found that AZM upregulates the expression of MDA5 and RIG-I, pathogen recognition receptors (PRRs) induced by ZIKV infection, and increases the levels of phosphorylated TBK1 and IRF3. Interestingly, AZM treatment upregulates phosphorylation of TBK1, without inducing phosphorylation of IRF3 by itself. These findings highlight the potential use of AZM as a broad antiviral agent to combat viral infection and prevent ZIKV associated devastating clinical outcomes, such as congenital microcephaly.

Copyright © 2019 American Society for Microbiology. All Rights Reserved.

Keywords: Antivirals; Antibiotics; Azithromycin; Zika Virus.

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#Growth activation of #influenza virus by #trypsin and effect of T-705 (#favipiravir) on trypsin-optimized growth condition (Acta Virol., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Acta Virol. 2019;63(3):309-315. doi: 10.4149/av_2019_311.

Growth activation of influenza virus by trypsin and effect of T-705 (favipiravir) on trypsin-optimized growth condition.

Daikoku T, Okuda T, Kawai M, Morita N, Tanaka T, Takemoto M, Fukuda Y, Takahashi K, Nomura N, Shiraki K.

 

Abstract

Influenza virus is activated by proteolytic cleavage of hemagglutinin by trypsin. After determining the optimal trypsin concentration, intracellular and extracellular influenza A/PR/8/34 (H1N1) and A/Victoria/361/2011 (H3N2) virus productions were compared in cultures treated with T-705 (favipiravir) and GS 4071 (an active form of oseltamivir). Although both drugs efficiently inhibited extracellular viral RNA release in a dose-dependent manner, T-705 inhibited it to the level of the inoculum without trypsin treatment, while GS 4071 inhibited it to a final level 10 times higher than that without trypsin. T-705 inhibited intracellular viral RNA production to the level of input virus in both trypsin-treated and untreated cells. In contrast, GS 4071 dose-dependently inhibited intracellular viral RNA production in cells treated with trypsin but allowed viral RNA synthesis. The level of maximum inhibition by GS 4071 was 10 times higher than that of cells without trypsin and 1,000 times greater than the inoculum titer in cells without trypsin. T-705 inhibited both intracellular and extracellular virus production 1,000 and 10 times more strongly, respectively, than GS 4071. T-705 has powerful anti-influenza activity in the absence of trypsin and even in the trypsin-optimized growth condition, suggesting the therapeutic advantage in treatment of influenza complicated with bacterial pneumonia.

Keywords: influenza; T-705; Tamiflu; trypsin; bacterial trypsin-like protease.

PMID: 31507197 DOI: 10.4149/av_2019_311

Keywords: Influenza A; H1N1; H3N2; Antivirals; Favipiravir; Oseltamivir.

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A study of the #relationship between #human #infection with #avian #influenza a (#H5N6) and environmental avian influenza viruses in #Fujian, #China (BMC Infect Dis., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

BMC Infect Dis. 2019 Sep 2;19(1):762. doi: 10.1186/s12879-019-4145-6.

A study of the relationship between human infection with avian influenza a (H5N6) and environmental avian influenza viruses in Fujian, China.

Chen P1, Xie JF1,2, Lin Q2, Zhao L2, Zhang YH2, Chen HB2, Weng YW1,2, Huang Z2, Zheng KC3,4.

Author information: 1 College of Public Health, Fujian Medical University, No. 88, Jiaotong Road, Taijiang District, Fuzhou, 350000, China. 2 Fujian Center for Disease Control and Prevention, Fujian Provincial Key Laboratory of Zoonosis Research, Fuzhou, 350001, China. 3 College of Public Health, Fujian Medical University, No. 88, Jiaotong Road, Taijiang District, Fuzhou, 350000, China. kingdadi9909@126.com. 4 Fujian Center for Disease Control and Prevention, Fujian Provincial Key Laboratory of Zoonosis Research, Fuzhou, 350001, China. kingdadi9909@126.com.

 

Abstract

BACKGROUND:

Avian influenza A (H5N6) virus poses a great threat to the human health since it is capable to cross the species barrier and infect humans. Although human infections are believed to largely originate from poultry contaminations, the transmissibility is unclear and only limited information was available on poultry environment contaminations, especially in Fujian Province.

METHODS:

A total of 4901 environmental samples were collected and tested for Avian Influenza Virus (AIV) from six cities in Fujian Province through the Fujian Influenza Surveillance System from 2013 to 2017. Two patient-related samples were taken from Fujian’s first confirmed H5N6 human case and his backyard chicken feces in 2017. Chi-square test or Fisher’s exact probability test was used to compare the AIV and the viral subtype positive rates among samples from different Surveillance cities, surveillance sites, sample types, and seasons. Phylogenetic tree analysis and molecular analysis were conducted to track the viral transmission route of the human infection and to map out the evolutions of H5N6 in Fujian.

RESULTS:

The overall positive rate of the H5 subtype AIVs was 4.24% (208/4903). There were distinctive differences (p < 0.05) in the positive rates in samples from different cities, sample sites, sample types and seasons. The viruses from the patient and his backyard chicken feces shared high homologies (99.9-100%) in all the eight gene segments. Phylogenetic trees also showed that these two H5N6 viruses were closely related to each other, and were classified into the same genetic clade 2.3.4.4 with another six H5N6 isolates from the environmental samples. The patient’s H5N6 virus carried genes from H6N6, H5N8 and H5N6 viruses originated from different areas. The R294K or N294S substitution was not detected in the neuraminidase (NA). The S31 N substitution in the matrix2 (M2) gene was detected but only in one strain from the environmental samples.

CONCLUSIONS:

The H5 subtype of AIVs has started circulating in the poultry environments in Fujian Province. The patient’s viral strain originated from the chicken feces in his backyard. Genetic reassortment in H5N6 viruses in Fujian Province was indicated. The H5N6 viruses currently circulating in Fujian Province were still commonly sensitive to Oseltamivir and Zanamivir, but the resistance against Amantadine has emerged.

KEYWORDS: Avian influenza a (H5N6) virus; Environmental contamination; Phylogenetic analysis

PMID: 31477028 DOI: 10.1186/s12879-019-4145-6

Keywords: Antivirals; Drugs Resistance; Oseltamivir; Zanamivir; Amantadine; H5N6; H6N6; H5N8; Reassortant strain; Avian Influenza; Human; Fujian; China.

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#Cholesterol-conjugated stapled #peptides inhibit #Ebola and #Marburg viruses in vitro and in vivo (Antiviral Res., abstract)

[Source: US National Library of Medicine, full page: (LINK). Abstract, edited.]

Antiviral Res. 2019 Aug 29:104592. doi: 10.1016/j.antiviral.2019.104592. [Epub ahead of print]

Cholesterol-conjugated stapled peptides inhibit Ebola and Marburg viruses in vitro and in vivo.

Pessi A1, Bixler SL2, Soloveva V2, Radoshitzky S2, Retterer C3, Kenny T3, Zamani R3, Gomba G3, Gharabeih D3, Wells J3, Wetzel KS3, Warren TK2, Donnelly G3, Van Tongeren SA3, Steffens J3, Duplantier AJ2, Kane CD4, Vicat P5, Couturier V5, Kester KE6, Shiver J6, Carter K7, Bavari S8.

Author information: 1 PeptiPharma, Rome, Italy. 2 United States Army Medical Research Institute of Infectious Diseases (USAMRIID), 1425 Porter Street, Frederick, MD, 21702, USA; Therapeutic Development Center, USAMRIID, Frederick, MD, 21702, USA. 3 United States Army Medical Research Institute of Infectious Diseases (USAMRIID), 1425 Porter Street, Frederick, MD, 21702, USA. 4 United States Army Medical Research Institute of Infectious Diseases (USAMRIID), 1425 Porter Street, Frederick, MD, 21702, USA; Therapeutic Development Center, USAMRIID, Frederick, MD, 21702, USA. Electronic address: christopher.d.kane.civ@mail.mil. 5 Sanofi, Paris, France. 6 Sanofi Pasteur, Swiftwater, PA, USA. 7 Sanofi, Cambridge, MA, USA. 8 United States Army Medical Research Institute of Infectious Diseases (USAMRIID), 1425 Porter Street, Frederick, MD, 21702, USA. Electronic address: sina.bavari.civ@mail.mil.

 

Abstract

The Filoviridae family consists of five ebolavirus species, Ebola (EBOV), Sudan (SUDV), Bundibugyo virus (BDBV), Reston (RESTV), and Taï Forest (TAFV) viruses, and a Marburgvirus species, with Marburg (MARV) and Ravn (RAVV) viruses. Ebola virus (EBOV) has emerged as a significant public health concern since the 2013-2016 Ebola Virus Disease outbreak in Western Africa. Currently, there are no therapeutics approved and the need for Ebola-specific therapeutics remains a gap. In search for anti-Ebola therapies we tested the idea of using inhibitory properties of peptides corresponding to the C-terminal heptad-repeat (HR2) domains of class I fusion proteins against EBOV infection. The fusion protein GP2 of EBOV belongs to class I, suggesting that a similar strategy to HIV may be applied to inhibit EBOV infection. The serum half-life of peptides was expanded by cholesterol conjugation to allow daily dosing. The peptides were further constrained to stabilize a helical structure to increase the potency of inhibition. The EC50s of lead peptides were in low micromolar range, as determined by a high-content imaging test of EBOV-infected cells. Lead peptides were tested in an EBOV lethal mouse model and efficacy of the peptides were determined following twice-daily administration of peptides for 9 days. The most potent peptide was able to protect mice from lethal challenge of mouse-adapted Ebola virus. These data show that engineered peptides coupled with cholesterol can inhibit viral production, protect mice against lethal EBOV infection, and may be used to build novel therapeutics against EBOV.

Copyright © 2019. Published by Elsevier B.V.

KEYWORDS: Cholesterol; Ebola virus; Peptides; Rodents; Therapeutic

PMID: 31473342 DOI: 10.1016/j.antiviral.2019.104592

Keywords: Ebola; Marburg; Antivirals; Animal models.

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Cryo-EM #structure of #pleconaril-resistant #rhinovirus-B5 complexed to the #antiviral OBR-5-340 reveals unexpected binding site (Proc Natl Acad Sci USA, abstract)

[Source: Proceedings of the National Academy of Sciences of the United States of America, full page: (LINK). Abstract, edited.]

Cryo-EM structure of pleconaril-resistant rhinovirus-B5 complexed to the antiviral OBR-5-340 reveals unexpected binding site

Jiri Wald, Marion Pasin, Martina Richter, Christin Walther, Neann Mathai, Johannes Kirchmair, Vadim A. Makarov, Nikolaus Goessweiner-Mohr, Thomas C. Marlovits, Irene Zanella, Antonio Real-Hohn, Nuria Verdaguer, Dieter Blaas, and Michaela Schmidtke

PNAS first published August 28, 2019 / DOI: https://doi.org/10.1073/pnas.1904732116

Edited by Peter Palese, Icahn School of Medicine at Mount Sinai, New York, NY, and approved August 1, 2019 (received for review March 29, 2019)

 

Significance

More than 160 rhinovirus (RV) types cause about a billion respiratory infections annually in the United States alone, contributing to influenza-like illness. This diversity makes vaccination impractical. Existing small-molecule inhibitors target RVs by binding to a hydrophobic pocket in the capsid but exhibit side effects, resistance, and/or mutational escape, impeding registration as drugs. The pyrazolopyrimidine OBR-5-340 acts like other capsid binders by preventing conformational changes required for genome release. However, by using cryo-EM, we show that OBR-5-340 inhibits the naturally pleconaril-resistant RV-B5 by attaching close to the pocket entrance in a binding geometry different from that of most capsid binders. Combinations of inhibitors with disparate binding modes might thus effectively combat RVs while reducing the risk of resistance development.

 

Abstract

Viral inhibitors, such as pleconaril and vapendavir, target conserved regions in the capsids of rhinoviruses (RVs) and enteroviruses (EVs) by binding to a hydrophobic pocket in viral capsid protein 1 (VP1). In resistant RVs and EVs, bulky residues in this pocket prevent their binding. However, recently developed pyrazolopyrimidines inhibit pleconaril-resistant RVs and EVs, and computational modeling has suggested that they also bind to the hydrophobic pocket in VP1. We studied the mechanism of inhibition of pleconaril-resistant RVs using RV-B5 (1 of the 7 naturally pleconaril-resistant rhinoviruses) and OBR-5-340, a bioavailable pyrazolopyrimidine with proven in vivo activity, and determined the 3D-structure of the protein-ligand complex to 3.6 Å with cryoelectron microscopy. Our data indicate that, similar to other capsid binders, OBR-5-340 induces thermostability and inhibits viral adsorption and uncoating. However, we found that OBR-5-340 attaches closer to the entrance of the pocket than most other capsid binders, whose viral complexes have been studied so far, showing only marginal overlaps of the attachment sites. Comparing the experimentally determined 3D structure with the control, RV-B5 incubated with solvent only and determined to 3.2 Å, revealed no gross conformational changes upon OBR-5-340 binding. The pocket of the naturally OBR-5-340-resistant RV-A89 likewise incubated with OBR-5-340 and solved to 2.9 Å was empty. Pyrazolopyrimidines have a rigid molecular scaffold and may thus be less affected by a loss of entropy upon binding. They interact with less-conserved regions than known capsid binders. Overall, pyrazolopyrimidines could be more suitable for the development of new, broadly active inhibitors.

rhinovirus – capsid binder – inhibitor – 3D structure – cryo-EM

 

Footnotes

1 J.W., M.P., M.R., and C.W. contributed equally to this work.

2 To whom correspondence may be addressed. Email: dieter.blaas@meduniwien.ac.at or michaela.schmidtke@med.uni-jena.de.

Author contributions: J.K., T.C.M., A.R-H., N.V., D.B., and M.S. designed research; J.K, T.C.M, D.B., and M.S. supervised research; J.W., M.P., M.R., C.W., N.M., J.K., I.Z., A.R-H., and D.B. performed research; V.A.M. contributed new reagents/analytic tools; J.W., M.P., M.R., C.W., N.M., J.K., N.G-M., A.R-H., D.B., and M.S. analyzed data; D.B. reconstructed and refined the cryo-EM data; M.P. reconstructed and refined the cryo-EM data and carried out the thermal stability assays; M.R., C.W., and M.S. carried out the antiviral tests; N.M. analyzed binding sites of all compounds; I.Z. carried out stability assays; N.V. critically assessed the manuscript; and N.M., J.K., D.B., and M.S. wrote the paper.

The authors declare no conflict of interest.

This article is a PNAS Direct Submission.

Data deposition: The 3D structures reported in this paper have been deposited in the Research Collaboratory for Structural Bioinformatics (RCSB) Protein Data Bank, http://www.rcsb.org/ and the Electron Microscopy Data Bank, https://www.ebi.ac.uk/pdbe/emdb/ (accession nos.: RV-B5 complexed to OBR-5-340, PDB 6SK5 and EMD-10220; RV-B5, PDB 6SK6 and EMD-10221; RV-A89, PDB 6SK7and EMD-10222).

This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1904732116/-/DCSupplemental.

Published under the PNAS license.

Keywords: Antivirals; Drugs Resistance; Pleconaril; Enterovirus; Rhinovirus.

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