In Vitro Selection and Characterization of Influenza A (A/N9) Virus Variants Resistant to a Novel Neuraminidase Inhibitor, A-315675

Molla, Akhteruzzaman; Kati, Warren M.; Carrick, Robert J.; Steffy, Kevin; Shi, Yan; Montgomery, Debra; Gusick, Nanette; Stoll, V.S.; Stewart, Kent D.; Ng, Teresa I.; Maring, Clarence J.; Kempf, Dale J.; Kohlbrenner, William E.

doi:10.1128/jvi.76.11.5380-5386.2002

Cited by 48 publications

(37 citation statements)

References 32 publications

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“…The variant with Val at 119 was resistant to oseltamivir carboxylate but retained susceptibility to the other three inhibitors ( Table 2, M8 and M9). In vitro selection has determined that resistance to zanamivir was conferred by replacement of Glu with Gly, and the variant retained susceptibility to the remaining inhibitors ( It was reported previously that a variant (A/N9) selected in cell culture in the presence of A-315675 acquired the substitution of Asp for Glu at 119, which led to a 355-fold reduction in drug susceptibility (32). In our study, the zanamivir-selected variant carrying this substitution in the enzyme of the N2 subtype exhibited approximately 50-and 30-fold increases in IC 50 values against A-315675 and peramivir, respectively (Table 1, M7).…”

Section: Discussionmentioning

confidence: 99%

Susceptibilities of Antiviral-Resistant Influenza Viruses to Novel Neuraminidase Inhibitors

Mishin

Hayden

Gubareva

2005

Antimicrob Agents Chemother

196

153

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The susceptibilities of five zanamivir-resistant and six oseltamivir-resistant influenza viruses were assessed against four neuraminidase (NA) inhibitors, including peramivir and A-315675, by a fluorometric NA activity inhibition assay. The enzyme activity of a majority of the variants was effectively inhibited by either A-315675 or both peramivir and A-315675 (50% inhibitory concentration, <10 nM). A novel oseltamivir-resistant influenza virus B variant carrying substitution at residue 198 (Asp3Asn) (N2 numbering) retained susceptibility to peramivir and A-315675. In vivo, the Asn198 variant showed no apparent fitness impairment as judged by its recovery on day 5 from the nasal washes of ferrets coinfected with equal doses of the wild-type virus and the Asn198 variant. Based on the sequence analysis of the virus in the nasal washes, oseltamivir treatment (5 mg/kg twice daily for 5 days) did not provide growth advantage to the Asn198 variant. Nevertheless, treatment with A-315675 (prodrug A-322278) reduced the number of the animals (two of seven) shedding the Asn198 variant. These studies indicate that different patterns of susceptibility and cross-resistance between NA inhibitors may prove important if antiviral resistance to zanamivir and oseltamivir were to emerge.

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Section: Discussionmentioning

confidence: 99%

Susceptibilities of Antiviral-Resistant Influenza Viruses to Novel Neuraminidase Inhibitors

Mishin

Hayden

Gubareva

2005

Antimicrob Agents Chemother

196

153

View full text Add to dashboard Cite

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“…However, the searching space is often limited by the defined number of clinical isolates being sequenced. In vitro selection or serial passaging could be employed for discovering beneficial mutations in virus (21), but it is limited by the stochasticity and frequency of spontaneous mutation. In addition, the selection process restricts the sensitivity to mutants that offer a minor beneficial effect, due to the highly competitive environment of selection.…”

Section: Discussionmentioning

confidence: 99%

“…To understand the dynamic nature of the influenza A mutations, phylogenetic analysis or in vitro selection has been commonly employed (16)(17)(18)(19)(20)(21)(22). However, the phylogenetic approach does not allow the evaluation of all possible mutations and has a prediction power restricted by dependency on preexisting clinical isolates.…”

mentioning

confidence: 99%

Systematic Identification of H274Y Compensatory Mutations in Influenza A Virus Neuraminidase by High-Throughput Screening

Young

Dandekar

et al. 2013

J Virol

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“…The search for better and more useful NA inhibitors is under way. The pyrrolidinebased compound A-315675, the pyrazinecarboximide-based compound T-750, and the cyclopentane peramivir (RWJ-270201) are under study for their activities against influenza type A and B viruses (1,12,26,27,38). Peramivir has in vitro and in vivo activities equal to or greater than those of zanamivir and oseltamivir carboxylate against a number of influenza type A and B viruses (2,4,5,10,18,(46)(47)(48)(49).…”

Section: Discussionmentioning

confidence: 99%

Phenotypic Drug Susceptibility Assay for Influenza Virus Neuraminidase Inhibitors

McSharry

McDonough

Olson

et al. 2004

Clin Vaccine Immunol

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RNA viruses, such as influenza virus, have a high rate of mutation. Some of these mutations lead to viruses that are resistant to the currently used antiviral drugs and can be selected in the presence of antiviral drugs. If the drug-resistant viruses are biofit, their replication can lead to serious disease that cannot be treated effectively with the previously used antiviral compounds. This scenario has occurred frequently. When amantadine hydrochloride was used to treat influenza virus type A infections, 30% of the virus isolates obtained from treated patients were found to be resistant (9,11,22). With the licensing of the neuraminidase (NA) inhibitors, the selection of influenza viruses resistant to these inhibitors was of concern (32, 39, 43, 52, 61). In vitro resistance associated with amino acid substitutions in the hemagglutinin (HA) or NA antigens or both has been reported for the NA inhibitors (4,14,15,32,40,49,55). Despite these concerns, recent reports have demonstrated that there is little or no natural resistance to oseltamivir or zanamivir (5, 33). To determine if mutations to zanamavir occurred in vivo, the drug susceptibilities of clinical isolates obtained during a phase II clinical trial of zanamivir were determined by the plaque reduction assay (PRA), the NA inhibition (NAI) assay, and an in vivo assay using ferrets (3,17). A comparison of 41 paired isolates obtained before and during therapy with zanamivir showed no shifts in susceptibility to zanamivir when measured by the NAI assay, but the PRA using MDCK cells showed variable susceptibility to zanamivir. The susceptibilities of the clinical isolates determined by the PRA did not correlate with in vivo susceptibility studies in humans and ferrets, whereas the NAI assay did correlate with the in vivo susceptibility assays. In a study of 54 isolates obtained after treatment with oseltamivir, 2 clinical isolates were resistant in the NAI assay and an additional 8 were resistant in the PRA (16). These discrepancies between the PRA and the NAI assay could be due to the isolation of viruses with mutations in the HA gene that lead to in vitro resistance. NA inhibitor-resistant viruses with mutations in the HA gene would be scored in the PRA, but not in the NAI assay. The close relationship between the drug susceptibilities obtained with the NAI assay and the in vivo assays suggests that for these clinical isolates the NAI assay correlates better with the in vivo assay than the PRA for the NA inhibitors. The present evidence suggests that only mutations in the NA gene that lead to resistance to the NA inhibitors are clinically relevant.The currently used in vitro drug susceptibility assays, such as the PRA, the virus yield reduction assay, and the neutral red dye uptake assay, are cumbersome, time-consuming, and subjective (21, 45). A PCR-based drug susceptibility assay has recently been published, but its usefulness in clinical trials has not been evaluated (54). Previously, we demonstrated that the susceptibilities of herpes simplex viruses and h...

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In Vitro Selection and Characterization of Influenza A (A/N9) Virus Variants Resistant to a Novel Neuraminidase Inhibitor, A-315675

Cited by 48 publications

References 32 publications

Susceptibilities of Antiviral-Resistant Influenza Viruses to Novel Neuraminidase Inhibitors

Susceptibilities of Antiviral-Resistant Influenza Viruses to Novel Neuraminidase Inhibitors

Systematic Identification of H274Y Compensatory Mutations in Influenza A Virus Neuraminidase by High-Throughput Screening

Phenotypic Drug Susceptibility Assay for Influenza Virus Neuraminidase Inhibitors

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