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Antiviral Res . High-throughput screening for identification of Influenza A inhibitors using a cell-based immunofluorescence assay

tetano

Editor, Senior Moderator
Antiviral Res


. 2025 Jun 6:106209.
doi: 10.1016/j.antiviral.2025.106209. Online ahead of print. High-throughput screening for identification of Influenza A inhibitors using a cell-based immunofluorescence assay

Yohanka Martinez-Gzegozewska[SUP] 1 [/SUP], Lynn Rasmussen[SUP] 2 [/SUP], N Miranda Nebane[SUP] 2 [/SUP], Sara McKellip[SUP] 2 [/SUP], Dee Radzieta[SUP] 2 [/SUP], Anna Manuvakhova[SUP] 2 [/SUP], Andrew J Reece[SUP] 2 [/SUP], Pedro Ruiz[SUP] 2 [/SUP], Sixue Zhang[SUP] 3 [/SUP], Omar Moukha-Chafiq[SUP] 3 [/SUP], Melinda Sosa[SUP] 2 [/SUP], Corinne Augelli-Szafran[SUP] 3 [/SUP], Richard Whitley[SUP] 4 [/SUP], Robert Bostwick[SUP] 2 [/SUP], Paige Vinson[SUP] 2 [/SUP]



Affiliations
Abstract

A highly sensitive cell-based imaging assay has been used to screen a 200,000 compounds library for potential influenza antivirals. Compounds were screened at a concentration of 10 μM against the influenza strain A/Udorn/72 (H3N2) and the duplex capability of the assay was taken advantage of to select compounds with no or low cytotoxicity. The assay was also used for confirmation in concentration-response of the active compounds. In the set of confirmed hits, three major structural clusters, 23 minor clusters and 86 singletons were identified. Further evaluation of anti-influenza activity was performed for two additional influenza strains of the H1N1 subtype, A/WSN/33 and A/California/07/09. Three compounds from cluster A, SRI-44211, SRI-44215 and SRI-44221 showed selectivity indices for the pandemic strain A/California/07/09 in the range of 54.3 to 252, providing evidence that the core structure of cluster A could play a relevant role in their inhibitory potency and may serve as a starting point for future hit-to-lead optimization efforts.

Keywords: Influenza; cell-based; cluster; duplex; high-throughput; immunofluorescence.

 
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