2. Academic Validation
  3. High-throughput screening for identification of influenza a inhibitors using a cell-based immunofluorescence assay

High-throughput screening for identification of influenza a inhibitors using a cell-based immunofluorescence assay

  • Antiviral Res. 2025 Aug:240:106209. doi: 10.1016/j.antiviral.2025.106209.
Yohanka Martinez-Gzegozewska 1,  Lynn Rasmussen 2,  N Miranda Nebane 2,  Sara McKellip 2,  Dee Radzieta 2,  Anna Manuvakhova 2,  Andrew J Reece 2,  Pedro Ruiz 2,  Sixue Zhang 3,  Omar Moukha-Chafiq 2,  Melinda Sosa 2,  Corinne Augelli-Szafran 2,  Richard Whitley 3,  Robert Bostwick 2,  Paige Vinson 2
Affiliations

Affiliations

  • 1 Southern Research, Scientific Platforms Division, 2000 9th Ave South, Birmingham, AL 35205, USA. Electronic address: [email protected].
  • 2 Southern Research, Scientific Platforms Division, 2000 9th Ave South, Birmingham, AL 35205, USA.
  • 3 University of Alabama at Birmingham, 1720 2nd Ave South, Birmingham, AL 35294, USA.
PMID: 40484321 DOI: 10.1016/j.antiviral.2025.106209
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 54.3-252, 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

Cell-based; Cluster; Duplex; High-throughput; Immunofluorescence; Influenza.

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