#Functional #neuraminidase #inhibitor #resistance motifs in #avian #influenza A(#H5Nx) viruses (Antiviral Res., abstract)

[Source: Antiviral Research, full page: (LINK). Abstract, edited.]

Antiviral Research | Available online 1 August 2020, 104886 | In Press, Journal Pre-proof | Research paper

Functional neuraminidase inhibitor resistance motifs in avian influenza A(H5Nx) viruses

Dagmara Bialy, Holly Shelton, The Pirbright Institute, Pirbright, United Kingdom

Received 24 March 2020, Revised 13 July 2020, Accepted 16 July 2020, Available online 1 August 2020.

DOI: https://doi.org/10.1016/j.antiviral.2020.104886

 

Highlights

  • A R292K residue change in NA reduces susceptibility to NA inhibitor drugs (NAIs) in H5N6 and H5N2 avian influenza viruses.
  • All four mutations (E119V, H274Y, R292K and N294S) reducing susceptibility to NAIs in H5N6 viruses also reduced viral NA activity.
  • Reduced susceptibility to NA inhibitors in H5N6 did not attenuate virus replication efficiency in chicken cells or eggs.
  • A reduction of the viral HA affinity for sialic acid was observed in H5N6 viruses with reduced NA activity.

 

Abstract

Neuraminidase inhibitors (NAIs) are antiviral agents recommended worldwide to treat or prevent influenza virus infections in humans. Past influenza virus pandemics seeded by zoonotic infection by avian influenza viruses (AIV) as well as the increasing number of human infections with AIV have shown the importance of having information about resistance to NAIs by avian NAs that could cross the species barrier. In this study we introduced four NAI resistance-associated mutations (N2 numbering) previously found in human infections into the NA of three current AIV subtypes of the H5Nx genotype that threaten the poultry industry and human health: highly pathogenic H5N8, H5N6 and H5N2. Using the established MUNANA assay we showed that a R292K substitution in H5N6 and H5N2 viruses significantly reduced susceptibility to three licenced NAIs: oseltamivir, zanamivir and peramivir. In contrast the mutations E119V, H274Y and N294S had more variable effects with NAI susceptibility being drug- and strain-specific. We measured the replicative fitness of NAI resistant H5N6 viruses and found that they replicated to comparable or significantly higher titres in primary chicken cells and in embryonated hens’ eggs as compared to wild type – despite the NA activity of the viral neuraminidase proteins being reduced. The R292K and N294S drug resistant H5N6 viruses had single amino acid substitutions in their haemagglutinin (HA): Y98F and A189T, respectively (H3 numbering) which reduced receptor binding properties possibly balancing the reduced NA activity seen. Our results demonstrate that the H5Nx viruses can support drug resistance mutations that confer reduced susceptibility to licenced NAIs and that these H5N6 viruses did not show diminished replicative fitness in avian cell cultures. Our results support the requirement for on-going surveillance of these strains in bird populations to include motifs associated with human drug resistance.

Keywords: Avian Influenza; H5N2; H5N6; Antivirals; Drugs Resistance; Oseltamivir; Zanamivir; Peramivir.

——

Characterization of #Neuraminidase #Inhibitor #Resistant #Influenza Virus Isolates From Immunocompromised Patients in the Republic of #Korea (Virol J., abstract)

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

Virol J. 2020 Jul 6;17(1):94. doi: 10.1186/s12985-020-01375-1.

Characterization of Neuraminidase Inhibitor-Resistant Influenza Virus Isolates From Immunocompromised Patients in the Republic of Korea

Heui Man Kim 1, Namjoo Lee 1, Mi-Seon Kim 1, Chun Kang 1, Yoon-Seok Chung 2

Affiliations: 1 Division of Viral Diseases, Center for Laboratory Control of Infectious Diseases, Korea Centers for Disease Control and Prevention, Cheongju-si, South Korea. 2 Division of Viral Diseases, Center for Laboratory Control of Infectious Diseases, Korea Centers for Disease Control and Prevention, Cheongju-si, South Korea. rollstone93@korea.kr.

PMID: 32631440 DOI: 10.1186/s12985-020-01375-1

 

Abstract

Background:

The emergence of influenza viruses resistant to anti-influenza drugs is a threat to global public health. The Korea Centers for Disease Control and Prevention operates the Korea Influenza and Respiratory Viruses Surveillance System (KINRESS) to monitor epidemics of influenza and Severe Acute Respiratory Infection (SARI) to identify mutated influenza viruses affecting drug resistance, pathogenesis, and transmission.

Methods:

Oropharyngeal swab samples were collected from KINRESS and SARI during the 2018-2019 season. The specimens confirmed influenza virus using real-time RT-PCR on inoculated MDCK cells. HA and NA sequences of the influenza viruses were analyzed for phylogeny and mutations. Neuraminidase inhibition and hemagglutination inhibition assays were utilized to characterize the isolates.

Results:

Two A(H1N1)pdm09 isolates harboring an H275Y substitution in the neuraminidase sequence were detected in patients with acute hematologic cancer. They had prolonged respiratory symptoms, with the virus present in the respiratory tract despite oseltamivir and peramivir treatment. Through the neuraminidase inhibition assay, both viruses were found to be resistant to oseltamivir and peramivir, but not to zanamivir. Although hemagglutinin and neuraminidase phylogenetic analyses suggested that the 2 A(H1N1)pdm09 isolates were not identical, their antigenicity was similar to that of the 2018-19 influenza vaccine virus.

Conclusions:

Our data indicate the utility of monitoring influenza-infected immunocompromised patients in general hospitals for the early detection of emerging neuraminidase inhibitor-resistant viruses and maintaining continuous laboratory surveillance of patients with influenza-like illness in sentinel clinics to monitor the spread of such new variants. Finally, characterization of the virus can inform the risk assessment for future epidemics and pandemics caused by drug-resistant influenza viruses.

Keywords: Drug resistance; H275Y; Immunocompromised patients; Influenza virus.

Keywords: Seasonal Influenza; H1N1pdm09; Cancer; Immunosuppression; Antivirals; Drugs Resistance; Oseltamivir; Zanamivir; Peramivir; S. Korea.

——

#Influenza #H1N1pdm09 virus exhibiting reduced #susceptibility to #baloxavir due to a PA E23K #substitution detected from a #child without baloxavir #treatment (Antiviral Res., abstract)

[Source: Antiviral Research, full page: (LINK). Abstract, edited.]

Antiviral Research | Available online 20 June 2020, 104828 | In Press, Journal Pre-proof | Short Communication

Influenza A(H1N1)pdm09 virus exhibiting reduced susceptibility to baloxavir due to a PA E23K substitution detected from a child without baloxavir treatment

Emi Takashita 1, Takashi Abe 2, Hiroko Morita 1, Shiho Nagata 1, Seiichiro Fujisaki 1, Hideka Miura 1, Masayuki Shirakura 1, Noriko Kishida 1, Kazuya Nakamura 1, Tomoko Kuwahara 1, KeikoMitamura 3, Masataka Ichikawa 4, Masahiko Yamazaki 5, Shinji Watanabe 1, Hideki Hasegawa 1, The Influenza Virus Surveillance Group of Japan,  Rika Komagome 6, Asami Ohnishi 7, Rika Tsutsui 8, Masaki Takahashi 9, MieSasaki 10, Shiho Tamura 11, Chihiro Shibata 12, Kenichi Komabayashi 13, Nozomi Saito 14, Aoi Saito 15, Fuminori Mizukoshi 16, Akira Wakatsuki 17, Hiroyuki Tsukagoshi 18, Noriko Suzuki 19, Yuka Uno 20, Noriko Oitate 21, Wakako Nishikawa  22, Mami Nagashima 23, Sumi Watanabe 24, Chiharu Kawakami 25, Hideaki Shimizu 26, Hazime Amano 27, Satoko Kanazawa 28, Kaori Watanabe 29, Kazunari Yamamoto 30, Tetsuya Yoneda 31, Sachiko Nakamura 32, Kaori Sato 33, Masayuki Oonuma 34, Michiko Takeuchi 35, ErinaTanaka 36, Masahiro Nishioka 37, Yusuke Sato 38, Yukiko Sakai 39, Takaharu Maehata 40, Toshihiko Furuta 41, Yoshihiro Yasui 42, Takuya Yano 43, Asa Tanino 44, Sachi Hirata 45, Akiko Nagasao 46, Satoshi Hiroi 47, Hideyuki Kubo 47, Fumika Okayama 48, Tomohiro Oshibe 49, Ai Mori 50, Ryutaro Murayama 51, Shoko Chiba 52, Yuki Matsui 53, Yuko Kiguchi 54, Koji Takeuchi 55, Tetsuo Mita 56, Kayoko Nomiya 57, Yukie Shimazu 58, Yoshiki Fujii 59, Shoichi Toda 60, Yumiko Kawakami 61, Yukari Terajima 62, Mayumi Yamashita 63, Tomiyo Takahashi 64, Yuki Ashizuka 65, Chinami Wasano 66, Takashi Kimura 67, Sanae Moroishi 68, Miho Urakawa 69, Takashi Sakai 70, Kaori Nishizawa 71, Toru Hayashi 72, Yu Matsuura 73, Yuka Hamada 74, Yumani Kuba 75

6 Hokkaido Institute of Public Health; 7 Sapporo City Institute of Public Health; 8 Aomori Prefectural Public Health and Environment Center; 9 Iwate Prefectural Research Institute for Environmental Sciences and Public Health; 10 Miyagi Prefectural Institute of Public Health and Environment; 11 Sendai City Institute of Public Health; 12 Akita Prefectural Research Center for Public Health and Environment; 13 Yamagata Prefectural Institute of Public Health; 14 Fukushima Prefectural Institute of Public Health; 15 Ibaraki Prefectural Institute of Public Health; 16 Tochigi Prefectural Institute of Public Health and Environmental Sciences; 17 Utsunomiya City Institute of Public Health and Environment Science; 18 Gunma Prefectural Institute of Public Health and Environmental Sciences; 19 Saitama Institute of Public Health; 20 Saitama City Institute of Health Science and Research;  21 Chiba Prefectural Institute of Public Health; 22 Chiba City Institute of Health and Environment; 23 Tokyo Metropolitan Institute of Public Health; 24 Kanagawa Prefectural Institute of Public Health; 25 Yokohama City Institute of Public Health; 26 Kawasaki City Institute of Public Health; 27 Yokosuka Institute of Public Health; 28 Sagamihara City Institute of Public Health; 29 Niigata Prefectural Institute of Public Health and Environmental Sciences; 30 Niigata City Institute of Public Health and Environment; 31 Toyama Institute of Health; 32 Ishikawa Prefectural Institute of Public Health and Environmental Science; 33 Fukui Prefectural Institute of Public Health and Environmental Science; 34 Yamanashi Institute for Public Health; 35 Nagano Environmental Conservation Research Institute; 36 Nagano City Health Center; 37
Gifu Prefectural Research Institute for Health and Environmental Sciences; 38 Gifu Municipal Institute of Public Health; 39 Shizuoka Institute of Environment and Hygiene; 40 Shizuoka City Institute of Environmental Sciences and Public Health; 41 Hamamatsu City Health Environment Research Center; 42 Aichi Prefectural Institute of Public Health; 43 Mie Prefecture Health and Environment Research Institute; 44 Shiga Prefectural Institute of Public Health; 45 Kyoto Prefectural Institute of Public Health and Environment; 46 Kyoto City Institute of Health and Environmental Sciences; 47 Osaka Institute of Public Health; 48 Sakai City Institute of Public Health; 49 Hyogo Prefectural Institute of Public Health Science; 50 Kobe Institute of Health; 51 Amagasaki City Institute of Public Health; 52 Nara Prefecture Institute of Health; 53 Wakayama Prefectural Research Center of Environment and Public Health; 54 Wakayama City Institute of Public Health; 55 Tottori Prefectural Institute of Public Health and Environmental Science; 56 Shimane Prefectural Institute of Public Health and Environmental Science; 57 Okayama Prefectural Institute for Environmental Science and Public Health; 58 Hiroshima Prefectural Technology Research Institute; 59 Hiroshima City Institute of Public Health; 60 Yamaguchi Prefectural Institute of Public Health and Environment; 61 Tokushima Prefectural Public Health, Pharmaceutical and Environmental Sciences Center; 62 Kagawa Prefectural Research Institute for Environmental Sciences and Public Health; 63 Ehime Prefecture Institute of Public Health and Environmental Science; 64 Kochi Public Health and Environmental Science Research Institute; 65 Fukuoka Institute of Health and Environmental Sciences; 66 Fukuoka City Institute of Health and Environment; 67 Kitakyushu City Institute of Health and Environmental Sciences; 68 Saga Prefectural Institute of Public Health and Pharmaceutical Research; 69 Nagasaki Prefectural Institute for Environment Research and Public Health; 70 Kumamoto Prefectural Institute of Public-Health and Environmental Science; 71 Kumamoto City Environmental Research Center; 72 Oita Prefectural Institute of Health and Environment; 73 Miyazaki Prefectural Institute for Public Health and Environment; 74 Kagoshima Prefectural Institute for Environmental Research and Public Health; 75 Okinawa Prefectural Institute of Health and Environment; 1 Influenza Virus Research Center, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama, Tokyo, 208-0011, Japan; 2 Abe Children’s Clinic, Minowa 2-15-22, Kohoku, Yokohama, Kanagawa, 223-0051, Japan; 3 Eiju General Hospital, Higashi Ueno 2-23-16, Taito, Tokyo, 110-8645, Japan; 4 Ichikawa Children’s Clinic, Higashi Odake 1544-3, Isehara, Kanagawa, 259-1133, Japan; 5 Zama Children’s Clinic, Tatsuno Dai 2-20-24, Zama, Kanagawa, 252-0023, Japan

Received 18 February 2020, Revised 14 May 2020, Accepted 28 May 2020, Available online 20 June 2020.

DOI: https://doi.org/10.1016/j.antiviral.2020.104828

 

Highlights

  • Influenza A(H1N1)pdm09 virus carrying a PA E23K substitution was detected.
  • The PA E23K mutant virus showed reduced baloxavir susceptibility.
  • The PA E23K mutant virus was isolated from a child without baloxavir treatment.
  • Possible transmission of the PA E23K mutant virus among humans is suggested.
  • Baloxavir susceptibility monitoring of influenza viruses is essential.

 

Abstract

Human-to-human transmission of PA I38 mutant influenza A(H3N2) viruses with reduced baloxavir susceptibility has been reported in Japan. In December 2019, we detected a PA E23K mutant A(H1N1)pdm09 virus from a child without baloxavir treatment. The PA E23K mutant virus exhibited reduced baloxavir susceptibility but remained susceptible to neuraminidase inhibitors. Epidemiological data suggest possible transmission of this PA E23K mutant virus among humans, although its growth capability relative to that of the wild-type virus was reduced. Therefore, baloxavir susceptibility monitoring of influenza viruses is essential.

Keywords: Influenza A; Antivirals; Drugs Resistance; Baloxavir; Oseltamivir; Pediatrics; Japan.

——

The #Antiviral Effects of #Baloxavir Marboxil Against #Influenza A Virus Infection in #Ferrets (Influenza Other Respir Viruses, abstract)

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

Influenza Other Respir Viruses. 2020 Jun 13. doi: 10.1111/irv.12760. Online ahead of print.

The Antiviral Effects of Baloxavir Marboxil Against Influenza A Virus Infection in Ferrets

Mitsutaka Kitano 1, Takanobu Matsuzaki 1, Ryoko Oka 1, Kaoru Baba 2, Takahiro Noda 2, Yuki Yoshida 1, Kenji Sato 1, Kohei Kiyota 1, Tohru Mizutare 1, Ryu Yoshida 1, Akihiko Sato 1, Hiroshi Kamimori 1, Takao Shishido 1, Akira Naito 1

Affiliations: 1 Shionogi & Co., Ltd., Toyonaka, Japan. 2 Shionogi TechnoAdvance Research, Co., Ltd., Toyonaka, Japan.

PMID: 32533654 DOI: 10.1111/irv.12760

 

Abstract

Background:

Baloxavir marboxil (BXM), the oral prodrug of baloxavir acid (BXA), greatly reduces virus titers as well as influenza symptoms of uncomplicated influenza in patients.

Objectives:

To investigate the pharmacokinetic profiles of BXA and its efficacy against influenza A virus infection in ferrets.

Methods:

Ferrets were dosed orally with BXM (10 and 30 mg/kg twice daily for 1 day), oseltamivir phosphate (OSP) (5 mg/kg twice daily for 2 days) or vehicle to measure the antiviral effects of BXM and OSP. The pharmacokinetic parameters of BXA was determined after single oral dosing of BXM.

Results:

The maximum plasma concentrations of BXA were observed at 1.50 and 2.00 hours with the two BXM doses, which then declined with an elimination half-life of 6.91 and 4.44 hours, respectively. BXM at both doses remained detectable in the plasma in ferrets, which may be due to higher stability in liver microsomes. BXM (10 and 30 mg/kg twice daily) treatment at Day 1 post-infection (p.i.) reduced virus titers by ≥3 log10 of the 50% tissue culture infective doses by Day 2, which was significantly different compared with vehicle or OSP. Body temperature drops over time were significantly greater with BXM than with vehicle or OSP. Significant reduction in virus titers was also demonstrated when BXM was administrated after symptom onset at Day 2 p.i. compared with vehicle and OSP, although body temperature changes largely overlapped between Day 2 and Day 4.

Conclusions:

The results highlight the rapid antiviral action of BXM with post-exposure prophylaxis or therapeutic dosing in ferrets and offer support for further research on prevention of influenza virus infection and transmission.

Keywords: baloxavir marboxil; ferrets; influenza A virus; pharmacodynamics; pharmacokinetics.

© 2020 The Authors. Influenza and Other Respiratory Viruses published by John Wiley & Sons Ltd.

Keywords: Influenza A; Antivirals; Baloxavir; Oseltamivir; Animal models.

—–

Host-targeted #Nitazoxanide has a high barrier to #resistance but does not reduce the emergence or #proliferation of #oseltamivir-resistant #influenza viruses in vitro or in vivo when used in combination with oseltamivir (Antiviral Res., abstract)

[Source: Antiviral Research, full page: (LINK). Abstract, edited.]

Antiviral Research | Available online 13 June 2020, 104851 | In Press, Journal Pre-proof | Research paper

Host-targeted Nitazoxanide has a high barrier to resistance but does not reduce the emergence or proliferation of oseltamivir-resistant influenza viruses in vitro or in vivo when used in combination with oseltamivir

Danielle Tilmanis 1, Paulina Koszalka 1, Ian G. Barr 1,3, Jean-Francois Rossignol 2, Edin Mifsud 1, Aeron C. Hurt 1,3

1 WHO Collaborating Centre for Reference and Research on Influenza, VIDRL, Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, 3000, Australia; 2 Romark Laboratories, L.C., Tampa, Florida, USA; 3 The University of Melbourne, Department of Microbiology and Immunology, Parkville, Victoria, 3010, Australia

Received 1 January 2020, Revised 5 June 2020, Accepted 7 June 2020, Available online 13 June 2020.

DOI: https://doi.org/10.1016/j.antiviral.2020.104851

 

Highlights

  • Serial passaging was used to determine the propensity for influenza viruses to develop resistance to tizoxanide.
  • Tizoxanide selective pressure up to 20 μM did not result in virus populations with altered drug susceptibility.
  • Host-targeted Nitazoxanide has a high barrier to antiviral resistance.
  • Tizoxanide/oseltamivir combination therapy did not prevent the emergence or selection of oseltamivir resistant virus.

 

Abstract

A major limitation of the currently available influenza antivirals is the potential development of drug resistance. The adamantanes, neuraminidase inhibitors, and more recently polymerase inhibitors, have all been associated with the emergence of viral resistance in preclinical, clinical studies or in clinical use. As a result, host-targeted drugs that act on cellular proteins or functions have become an attractive option for influenza treatment as they are less likely to select for resistance. Nitazoxanide (NTZ) is a host-targeted antiviral that is currently in Phase III clinical trials for the treatment of influenza. In this study, we investigated the propensity for circulating influenza viruses to develop resistance to nitazoxanide in vitro by serially passaging viruses under selective pressure. Phenotypic and genotypic analysis of viruses passaged ten times in the presence of up to 20 μM tizoxanide (TIZ; the active metabolite of nitazoxanide) showed that none had a significant change in TIZ susceptibility, and amino acid substitutions arising that were unique to TIZ passaged viruses, did not alter TIZ susceptibility.

Combination therapy, particularly utilising drugs with different mechanisms of action, is another option for combatting antiviral resistance, and while combination therapy has been shown to improve antiviral effects, the effect of reducing the emergence and selection of drug-resistant virus has been less widely investigated. Here we examined the use of TIZ in combination with oseltamivir, both in vitro and using the ferret model for influenza infection and found that the combination of the two drugs did not provide significant benefit in reducing the emergence or selection of oseltamivir-resistant virus.

These in vitro findings suggest that clinical use of NTZ may be significantly less likely to select for resistance in circulating influenza viruses compared to virus-targeted antivirals, and although the combination of NTZ with oseltamivir did not reduce the emergence of oseltamivir-resistant virus in vitro or in vivo, combination therapy with NTZ and other newer classes of influenza antiviral drugs should be considered due to NTZ’s higher host-based barrier to resistance.

View full text© 2020 Elsevier B.V. All rights reserved.

Keywords: Influenza A; Antivirals; Drugs Resistance; Nitazoxamide, Oseltamivir; Animal models.

—–

#Genotyping and #Reassortment #Analysis of Highly Pathogenic #Avian #Influenza Viruses #H5N8 and #H5N2 From #Egypt Reveals Successive Annual #Replacement of Genotypes (Infect Genet Evol., abstract)

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

Infect Genet Evol. 2020 May 23;104375. doi: 10.1016/j.meegid.2020.104375. Online ahead of print.

Genotyping and Reassortment Analysis of Highly Pathogenic Avian Influenza Viruses H5N8 and H5N2 From Egypt Reveals Successive Annual Replacement of Genotypes

Kareem E Hassan 1, Noha Saad 2, Hassanein H Abozeid 3, Salama Shany 4, Magdy F El-Kady 4, Abdelsatar Arafa 2, Azza A A El-Sawah 4, Florian Pfaff 5, Hafez M Hafez 6, Martin Beer 5, Timm Harder 7

Affiliations: 1 Institute of Diagnostic Virology, Friedrich-Loeffler-Institute, Greifswald, Riems, Germany; Department of Poultry Diseases, Faculty of Veterinary Medicine, Beni-Suef University, Beni-Suef 62511, Egypt. 2 National Laboratory for Veterinary Quality Control on Poultry Production, Animal Health Research Institute, 12618, Dokki, Giza, Egypt. 3 Department of Poultry Diseases, Faculty of Veterinary Medicine, Cairo University, Egypt. 4 Department of Poultry Diseases, Faculty of Veterinary Medicine, Beni-Suef University, Beni-Suef 62511, Egypt. 5 Institute of Diagnostic Virology, Friedrich-Loeffler-Institute, Greifswald, Riems, Germany. 6 Institute of Poultry Diseases, Free University Berlin, Berlin, Germany. 7 Institute of Diagnostic Virology, Friedrich-Loeffler-Institute, Greifswald, Riems, Germany. Electronic address: Timm.Harder@fli.de.

PMID: 32454245 DOI: 10.1016/j.meegid.2020.104375

 

Abstract

Highly pathogenic (HP) H5N1, clade 2.2.1, and low pathogenic avian influenza (LPAI) H9N2 viruses, G1-B lineage, are endemic in poultry in Egypt and have co-circulated for almost a decade. Surprisingly, no inter-subtypic reassortment events have been reported from the field during that time. After the introduction of HPAIV H5N8, clade 2.3.4.4b, in Egyptian poultry in 2016, suddenly HP H5N2 reassortants with H9N2 viruses emerged. The current analyses focussed on studying 32 duck flocks, 4 broiler chicken flocks, and 1 turkey flock, suffering from respiratory manifestations with moderate to high mortality reared in two Egyptian governorates during 2019. Real-time RT-PCR substantiated the presence of HP H5N8 in 21 of the 37 investigated flocks with mixed infection of H9N2 in two of them. HP H5N1 was not detected. Full hemagglutinin (HA) sequencing of 10 samples with full-genome sequencing of three of them revealed presence of a single genotype. Very few substituting mutations in the HA protein were detected versus previous Egyptian HA sequences of that clade. Interestingly, amino acid substitutions in the Matrix (M2) and the Neuraminidase (NA) proteins associated with conferring both Amantadine and Oseltamivir resistance were present. Systematic reassortment analysis of all publicly available Egyptian whole genome sequences of HP H5N8 (n = 23), reassortant HP H5N2 (n = 2) and LP H9N2 (n = 53) viruses revealed presence of at least seven different genotypes of HPAI H5Nx viruses of clade 2.3.4.4b in Egypt since 2016. For H9N2 viruses, at least three genotypes were distinguishable. Heat mapping and tanglegram analyses suggested that several internal gene segments in both HP H5Nx and H9N2 viruses originated from avian influenza viruses circulating in wild bird species in Egypt. Based on the limited set of whole genome sequences available, annual replacement patterns of HP H5Nx genotypes emerged and suggested selective advantages of certain genotypes since 2016.

Keywords: Beast analysis; Egypt; Genotyping; H5N2; H9N2; Highly pathogenic avian influenza; Phylogenetic analysis; Reassortment; Subtype H5N8; Tanglegram.

Copyright © 2019. Published by Elsevier B.V.

Conflict of interest statement. Declaration of Competing Interest: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Keywords: Avian Influenza; H5N2; H5N8; H9N2; Reassortant strain; Poultry; Wild Birds; Egypt; Antivirals; Drugs Resistance; Amantadine; Oseltamivir.

——

A phase 2 randomized, double-blind, placebo-controlled trial of MHAA4549A, a #mAb, plus #oseltamivir in patients hospitalized with #severe #influenza A infection (Antimicrob Agents Chemother., abstract)

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

A phase 2 randomized, double-blind, placebo-controlled trial of MHAA4549A, a monoclonal antibody, plus oseltamivir in patients hospitalized with severe influenza A infection

Jeremy J Lim, Anna C Nilsson, Michael Silverman, Nimer Assy, Priya Kulkarni, Jacqueline M McBride, Rong Deng, Chloe Li, Xiaoying Yang, Allen Nguyen, Priscilla Horn, Mauricio Maia, Aide Castro, Melicent C Peck, Joshua Galanter, Tom Chu, Elizabeth M Newton, Jorge A Tavel

DOI: 10.1128/AAC.00352-20

 

ABSTRACT

Background.

For patients hospitalized with severe influenza A, morbidity and mortality remain high. MHAA4549A, a human monoclonal antibody targeting the influenza A hemagglutinin stalk, has demonstrated pharmacological activity in animal studies and in a human influenza A challenge study. We evaluated the safety and efficacy of MHAA4549A plus oseltamivir against influenza A infection in hospitalized patients.

Methods.

The CRANE trial was a phase 2b, randomized, double-blind, placebo-controlled study of single intravenous (IV) doses of placebo, 3600-mg, or 8400-mg MHAA4549A together with oral oseltamivir (+OTV), in patients hospitalized with severe influenza A. Patients, enrolled across 68 clinical sites in 18 countries, were randomized 1:1:1. The primary outcome was the median time to normalization of respiratory function defined as the time to removal of supplemental oxygen support to maintain a stable SpO2 ≥ 95%. Safety, pharmacokinetics, and effects on influenza viral load were also assessed.

Results.

166 patients were randomized and analyzed during a preplanned interim analysis. Compared to placebo+OTV, MHAA4549A+OTV did not significantly reduce the time to normalization of respiratory function (placebo+OTV: 4.28 days; 3600-mg MHAA4549A+OTV: 2.78 days; 8400-mg MHAA4549A+OTV: 2.65 days), nor did it improve other secondary clinical outcomes. Adverse event frequency was balanced across cohorts. MHAA4549A+OTV did not further reduce viral load versus placebo+OTV.

Conclusions.

In hospitalized patients with influenza A, MHAA4549A did not improve clinical outcomes over OTV alone. Variability in patient removal from oxygen supplementation limited the utility of the primary endpoint. Validated endpoints are needed to assess novel treatments for severe influenza A.

Copyright © 2020 Lim et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

Keywords: Influenza A; Antivirals; Monoclonal antibodies; Oseltamivir.

——

A #Patient With #COVID19 Disease in a Referral #Hospital in #Iran: A Typical Case (Infect Disord Drug Targets, abstract)

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

Infect Disord Drug Targets. 2020 Apr 29. doi: 10.2174/1871526520666200429115535. Online ahead of print.

A Patient With COVID-19 Disease in a Referral Hospital in Iran: A Typical Case

Fereshteh Ghiasvand 1, Sepideh Zahak Miandoab 2, Hamid Harandi 3, Fereshteh Shahmari Golestan 4, Seyed Ahmad Seyed Alinaghi 3

Affiliations: 1 Liver Transplantation Research Center, Department of Infectious Diseases, Imam Khomeini Hospital Complex, Tehran University of Medical Sciences, Tehran. Iran. 2 Department of Infectious Diseases, Imam Khomeini Hospital Complex, Tehran University of Medical Sciences, Tehran. Iran. 3 Iranian Research Center for HIV/AIDS, Iranian Institute for Reduction of High Risk Behaviors, Tehran University of Medical Sciences, Tehran. Iran. 4 Imam Khomeini Hospital Complex, Tehran University of Medical Sciences, Tehran. Iran.

PMID: 32348232 DOI: 10.2174/1871526520666200429115535

 

Abstract

After the initial Wuhan outbreak in the end of December 2019, many new cases were found in other provinces of China and also many other countries over the world, including South Korea, Italy, Iran, Japan, and 68 other countries. We presented a 61-year-old woman with a history of diabetes mellitus was referred to the emergency department of a referral hospital in Tehran, Iran. The patient presented with fever, chills, and myalgia within three days. Laboratory analysis showed increased levels of erythrocyte sedimentation rate (ESR), and mild leukopenia. SARSCoV-2 PCR test -under Iran Ministry of Health and Medical Education (MoH&ME) guidelines – result was positive and the chest X-ray shows bilateral ground glass opacity. O2 saturation was 87% (without O2 therapy). The patient was hospitalized and treated with Oseltamivir 75 mg every 12 hours, Lopinavir/Ritonavir (Kaletra) 400/100 mg every 12 hours and hydroxychloroquine 400 mg stat. The patient last O2 saturation measured was 93% and had no fever in the 10th day of hospitalization. So she has been discharged from hospital and homeisolated according to Iran Ministry of Health protocol.

Keywords: COVID-19; Coronavirus; Diabetes mellitus; Ground glass Opacity; SARS-CoV-2 PCR test; severe acute respiratory syndrome coronavirus 2.

Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.

Keywords: SARS-CoV-2; COVID-19; Iran; Antivirals; Oseltamivir; Lopinavir; Ritonavir; Chloroquine; Diabetes.

—–

Efficacy of #neuraminidase #inhibitors against #H5N6 highly pathogenic #avian #influenza virus in a non-human #primate model (Antimicrob Agents Chemother., abstract)

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

Efficacy of neuraminidase inhibitors against H5N6 highly pathogenic avian influenza virus in a non-human primate model

Cong Thanh Nguyen, Saori Suzuki, Yasushi Itoh, Hirohito Ishigaki, Misako Nakayama, Kaori Hayashi, Keita Matsuno, Masatoshi Okamatsu, Yoshihiro Sakoda, Hiroshi Kida, Kazumasa Ogasawara

DOI: 10.1128/AAC.02561-19

 

ABSTRACT

Attention has been paid to H5N6 highly pathogenic avian influenza virus (HPAIV) because of its heavy burden on the poultry industry and human mortality. Since an influenza A virus carrying N6 neuraminidase (NA) has never spread in humans, the potential for H5N6 HPAIV to cause disease in humans and the efficacy of antiviral drugs against the virus need to be urgently assessed. We used non-human primates to elucidate the pathogenesis of H5N6 HPAIV as well as to determine the efficacy of antiviral drugs against the virus. H5N6 HPAIV infection led to high fever in cynomolgus macaques. The lung injury caused by the virus was severe with diffuse alveolar damage and neutrophil infiltration. In addition, an increase in IFN-α showed an inverse correlation with virus titers during the infection process. Oseltamivir was effective for reducing H5N6 HPAIV propagation, and continuous treatment with peramivir reduced virus propagation and severity of symptoms in the early stage. This study also showed the pathologically severe lung injury states in the cynomolgus macaques infected with H5N6 HPAIV, even in those that received early antiviral drug treatments, indicating the need for close monitoring and the need for further studies on the virus pathogenicity and new antiviral therapies.

Copyright © 2020 American Society for Microbiology. All Rights Reserved.

Keywords: Avian Influenza; H5N6; Antivirals; Oseltamivir; Peramivir; Animal models.

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#Computational Studies of #Drug #Repurposing and #Synergism of #Lopinavir, #Oseltamivir and #Ritonavir Binding With #SARS-CoV-2 Protease Against #COVID19 (J Biomol Struct Dyn., abstract)

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

J Biomol Struct Dyn, 1-7 2020 Apr 6 [Online ahead of print]

Computational Studies of Drug Repurposing and Synergism of Lopinavir, Oseltamivir and Ritonavir Binding With SARS-CoV-2 Protease Against COVID-19

Nisha Muralidharan 1, R Sakthivel 1, D Velmurugan 2, M Michael Gromiha 1

Affiliations: 1 Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai 600036, India. 2 School of Bioengineering, Department of Biotechnology, SRM University, Chennai, India.

PMID: 32248766 DOI: 10.1080/07391102.2020.1752802

 

Abstract

A novel coronavirus (SARS-CoV-2) has caused a major outbreak in humans all over the world, and it is the latest pandemic in the series of other infectious diseases. The concept of drug repurposing has been used successfully for many years for known diseases. Considering the emergency and urgency, drug repurposing concept is being explored for coronavirus disease (COVID-19) as well. Recently, the combination of three known drugs, lopinavir, oseltamivir and ritonavir has been proposed to control the virulence to a great extent in COVID-19 affected patients within 48 hours. Hence, we tried to understand the effect of synergism of these drugs against the SARS-CoV-2 protease using sequential docking studies. As a result, combination of three drugs showed a better binding energy than that of individual drugs. Further, the complex was subjected to molecular dynamics simulations to get insights into the stability of the complex, considering the simultaneous interactions between three drugs and the protein. The protein complexed with three drugs remained stable during the simulations. Hence, these drugs can be explored further for drug repurposing against the successful inhibition of COVID-19.

Keywords: COVID-19; SARS-CoV-2; drug repurposing; molecular docking; molecular dynamics simulations; protease.

Keywords: SARS-CoV-2; COVID-19; Antivirals; Oseltamivir; Ritonavir; Lopinavir.

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