Zanamivir–Cholesterol Conjugate: A Long-Acting Neuraminidase Inhibitor with Potent Efficacy against Drug-Resistant Influenza Viruses

Defining functional neuraminidase inhibitor drug resistance motifs in avian influenza viruses and the consequential impact on virus fitness in chicken cells

10.1101/663872 ◽

2019 ◽

Author(s):

Dagmara Bialy ◽

Holly Shelton

Keyword(s):

Drug Resistance ◽

Amino Acid ◽

Avian Influenza ◽

Neuraminidase Inhibitor ◽

Influenza Viruses ◽

Single Amino Acid ◽

Drug Resistant ◽

Human Influenza ◽

Wild Type ◽

Avian Influenza Viruses

ABSTRACTNeuraminidase inhibitors (NAIs) are antiviral agents recommended worldwide to treat or prevent influenza virus infections in humans. Mapping of functional resistance to currently licensed NAIs has been limited to human influenza viruses with only sporadic reports investigating avian influenza viruses (AIV). However past pandemics as well as the increasing number of humans infected with AIV have shown the importance of having information about avian NAs that could cross the species barrier. In this study we introduced four NAI resistance-associated mutations previously found in human strains into the NA of six prevalent AIV subtypes that threaten the poultry industry and human health: H7N9, H6N1, H4N6 and highly pathogenic H5N8, H5N6 and H5N2. Using the established MUNANA assay we show that R292K substitution significantly impaired NA activity in all strains, whereas E119V, H274Y and N294S have more variable effects on NA activity. The impact of these mutations on NAI susceptibility was drug- and strain-specific. We have shown that despite compromised NA activity drug-resistant H5N6 and H6N1 viruses replicated to comparable or significantly higher titres in primary chicken cells as compared to wild type. The replicative fitness of NAI-resistant H5N6 was also confirmedin ovo. Two drug resistant H5N6 viruses had single amino acid substitutions in their haemagglutinin (HA) which reduced receptor binding properties. Our results demonstrate that there are no universal NAI resistance determinants for all strains and although some are clearly deleterious for the virus, others can be rapidly compensated by acquiring concurrent changes in other gene segments.IMPORTANCEThe number of human infections caused by avian influenza viruses (AIV) keeps increasing. This together with the rapid emergence of influenza strains resistant to neuraminidase inhibitor drugs (NAIs) observed in the past raises a significant concern to public health. We studied the NAI resistance-associated molecular changes, previously reported in neuraminidase (NA) of human influenza, in AIV background. We found that single amino acid substitution can confer a multidrug resistance, or lead to a single-drug resistance across multiple virus subtypes. We also found that the drug-resistant viruses retained or showed enhanced fitness properties as compared to the corresponding wild-type, and this could be achieved by quick acquisition of concurrent mutations in haemagglutinin. Our study highlights the need for constant monitoring of NAI-resistance in AIV and understanding the molecular basis of antiviral resistance, as such information would be invaluable for pandemic preparedness and may facilitate the development of novel therapeutics.

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CS-8958, a Prodrug of the New Neuraminidase Inhibitor R-125489, Shows Long-Acting Anti-Influenza Virus Activity

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00333-08 ◽

2008 ◽

Vol 53 (1) ◽

pp. 186-192 ◽

Cited By ~ 152

Author(s):

Makoto Yamashita ◽

Takanori Tomozawa ◽

Masayo Kakuta ◽

Akane Tokumitsu ◽

Hatsumi Nasu ◽

...

Keyword(s):

Influenza Virus ◽

Acyl Chain ◽

Neuraminidase Inhibitor ◽

Influenza Viruses ◽

Infection Model ◽

Acyl Chain Length ◽

Long Acting ◽

Survival Effect ◽

First Time

ABSTRACT Two neuraminidase (NA) inhibitors, zanamivir (Relenza) and oseltamivir phosphate (Tamiflu), have been licensed for the treatment of and prophylaxis against influenza. In this paper, the new potent NA inhibitor R-125489 is reported for the first time. R-125489 inhibited the NA activities of various type A and B influenza viruses, including subtypes N1 to N9 and oseltamivir-resistant viruses. The survival effect of R-125489 was shown to be similar to that of zanamivir when administered intranasally in a mouse influenza virus A/Puerto Rico/8/34 infection model. Moreover, we found that the esterified form of R-125489 showed improved efficacy compared to R-125489 and zanamivir, depending on the acyl chain length, and that 3-(O)-octanoyl R-125489 (CS-8958) was the best compound in terms of its life-prolonging effect (P < 0.0001, compared to zanamivir) in the same infection model. A prolonged survival effect was observed after a single administration of CS-8958, even if it was given 7 days before infection. It is suggested that intranasally administered CS-8958 works as a long-acting NA inhibitor and shows in vivo efficacy as a result of a single intranasal administration.

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Punicalagin is a neuraminidase inhibitor of influenza viruses

Journal of Medical Virology ◽

10.1002/jmv.26449 ◽

2020 ◽

Author(s):

Ping Li ◽

Ruikun Du ◽

Zinuo Chen ◽

Yanyan Wang ◽

Peng Zhan ◽

...

Keyword(s):

Neuraminidase Inhibitor ◽

Influenza Viruses

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Antiviral Activity of Benzoic Acid Derivative NC-5 Against Influenza A Virus and Its Neuraminidase Inhibition

International Journal of Molecular Sciences ◽

10.3390/ijms20246261 ◽

2019 ◽

Vol 20 (24) ◽

pp. 6261

Author(s):

Min Guo ◽

Jiawei Ni ◽

Jie Yu ◽

Jing Jin ◽

Lingman Ma ◽

...

Keyword(s):

Antiviral Activity ◽

Benzoic Acid ◽

Influenza A Virus ◽

Influenza A ◽

Matrix Protein ◽

Influenza Viruses ◽

Drug Candidate ◽

Dependent Manner ◽

Drug Resistant

The currently available drugs against influenza A virus primarily target neuraminidase (NA) or the matrix protein 2 (M2) ion channel. The emergence of drug-resistant viruses requires the development of new antiviral chemicals. Our study applied a cell-based approach to evaluate the antiviral activity of a series of newly synthesized benzoic acid derivatives, and 4-(2,2-Bis(hydroxymethyl)-5-oxopyrrolidin-l-yl)-3-(5-cyclohexyl-4H-1,2,4-triazol-3-yl)amino). benzoic acid, termed NC-5, was found to possess antiviral activity. NC-5 inhibited influenza A viruses A/FM/1/47 (H1N1), A/Beijing/32/92 (H3N2) and oseltamivir-resistant mutant A/FM/1/47-H275Y (H1N1-H275Y) in a dose-dependent manner. The 50% effective concentrations (EC50) for H1N1 and H1N1-H275Y were 33.6 μM and 32.8 μM, respectively, which showed that NC-5 had a great advantage over oseltamivir in drug-resistant virus infections. The 50% cytotoxic concentration (CC50) of NC-5 was greater than 640 μM. Orally administered NC-5 protected mice infected with H1N1 and H1N1-H275Y, conferring 80% and 60% survival at 100 mg/kg/d, reducing body weight loss, and alleviating virus-induced lung injury. NC-5 could suppress NP and M1 protein expression levels during the late stages of viral biosynthesis and inhibit NA activity, which may influence virus release. Our study proved that NC-5 has potent anti-influenza activity in vivo and in vitro, meaning that it could be regarded as a promising drug candidate to treat infection with influenza viruses, including oseltamivir-resistant viruses.

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Factors Affecting Susceptibility of H5N1 Influenza Viruses to Neuraminidase Inhibitor Oseltamivir

Antiviral Research ◽

10.1016/j.antiviral.2008.01.022 ◽

2008 ◽

Vol 78 (2) ◽

pp. A18-A19

Author(s):

Elena Govorkova ◽

Natalia Ilyushina ◽

Jennifer McClaren ◽

Tri Naipospos ◽

Neziha Yilmas ◽

...

Keyword(s):

Neuraminidase Inhibitor ◽

Influenza Viruses ◽

Factors Affecting ◽

H5n1 Influenza

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Neuraminidase inhibitor resistance in influenza viruses

Journal of Medical Virology ◽

10.1002/jmv.20951 ◽

2007 ◽

Vol 79 (10) ◽

pp. 1577-1586 ◽

Cited By ~ 63

Author(s):

Phillip Andrew Reece

Keyword(s):

Neuraminidase Inhibitor ◽

Influenza Viruses ◽

Inhibitor Resistance

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Long-Acting Rilpivirine (RPV) Preexposure Prophylaxis Does Not Inhibit Vaginal Transmission of RPV-Resistant HIV-1 or Select for High-Frequency Drug Resistance in Humanized Mice

Journal of Virology ◽

10.1128/jvi.01912-19 ◽

2020 ◽

Vol 94 (8) ◽

Cited By ~ 1

Author(s):

Kevin Melody ◽

Chandra N. Roy ◽

Christopher Kline ◽

Mackenzie L. Cottrell ◽

Dwayne Evans ◽

...

Keyword(s):

High Frequency ◽

Low Frequency ◽

Humanized Mice ◽

Drug Resistant ◽

Low Level ◽

Preexposure Prophylaxis ◽

Vaginal Transmission ◽

Long Acting ◽

Hiv 1 ◽

Level Resistance

ABSTRACT As a long-acting formulation of the nonnucleoside reverse transcriptase inhibitor rilpivirine (RPV LA) has been proposed for use as preexposure prophylaxis (PrEP) and the prevalence of transmitted RPV-resistant viruses can be relatively high, we evaluated the efficacy of RPV LA to inhibit vaginal transmission of RPV-resistant HIV-1 in humanized mice. Vaginal challenges of wild-type (WT), Y181C, and Y181V HIV-1 were performed in mice left untreated or after RPV PrEP. Plasma viremia was measured for 7 to 10 weeks, and single-genome sequencing was performed on plasma HIV-1 RNA in mice infected during PrEP. RPV LA significantly prevented vaginal transmission of WT HIV-1 and Y181C HIV-1, which is 3-fold resistant to RPV. However, it did not prevent transmission of Y181V HIV-1, which has 30-fold RPV resistance in the viruses used for this study. RPV LA did delay WT HIV-1 dissemination in infected animals until genital and plasma RPV concentrations waned. Animals that became infected despite RPV LA PrEP did not acquire new RPV-resistant mutations above frequencies in untreated mice or untreated people living with HIV-1, and the mutations detected conferred low-level resistance. These data suggest that high, sustained concentrations of RPV were required to inhibit vaginal transmission of HIV-1 with little or no resistance to RPV but could not inhibit virus with high resistance. HIV-1 did not develop high-level or high-frequency RPV resistance in the majority of mice infected after RPV LA treatment. However, the impact of low-frequency RPV resistance on virologic outcome during subsequent antiretroviral therapy still is unclear. IMPORTANCE The antiretroviral drug rilpivirine was developed into a long-acting formulation (RPV LA) to improve adherence for preexposure prophylaxis (PrEP) to prevent HIV-1 transmission. A concern is that RPV LA will not inhibit transmission of drug-resistant HIV-1 and may select for drug-resistant virus. In female humanized mice, we found that RPV LA inhibited vaginal transmission of WT or 3-fold RPV-resistant HIV-1 but not virus with 30-fold RPV resistance. In animals that became infected despite RPV LA PrEP, WT HIV-1 dissemination was delayed until genital and plasma RPV concentrations waned. RPV resistance was detected at similar low frequencies in untreated and PrEP-treated mice that became infected. These results indicate the importance of maintaining RPV at a sustained threshold after virus exposure to prevent dissemination of HIV-1 after vaginal infection and low-frequency resistance mutations conferred low-level resistance, suggesting that RPV resistance is difficult to develop after HIV-1 infection during RPV LA PrEP.

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