Fig. 4: ISM3312 exhibits potent antiviral activity against various coronaviruses in human proximal airway organoid model.

Comparison of the antiviral efficacy of ISM3312, Nimatrelvir, and Ensitrelvir against SARS-CoV-2 BA.2.3 was evaluated in 2D human proximal airway organoids. Viral RNA of supernatants from the apical and basolateral chambers of the 2D proximal airway organoids was quantified by real-time PCR. Representative data from independent experiments utilizing organoids from three different donors were displayed (a), n = 3 biological replicates. Organoids were stained with anti-NP (green) and DAPI (blue). Representative immunofluorescence images of SARS-CoV-2 infected organoids at 72 h post-infection (b), with a scale bar indicating 50 μm. Anti-MERS-CoV activity measurement of ISM3312 in 2D Airway Organoids. Viral RNA in the apical and basolateral chamber supernatants were quantified by qRT–PCR (c), n = 3 biological replicates. Representative immunofluorescence images of MERS-CoV infected organoids at 72 h post-infection (d), with a scale bar indicating 50 μm. e–g Viral RNA gene copy number were detected to evaluate antiviral efficacy of ISM3312 against 229E, NL63, OC43, and HKU1 in 2D airway organoids at two concentrations (20 nM, 200 nM). 229E, NL63, and HKU1 were performed with three biological replicates, while OC43 was performed with three biological samples and three technical replicates. Representative immunofluorescence images were shown in f and g. Scale bar = 50 μm. Data were analyzed by using two-way ANOVA followed by Tukey’s multiple comparisons test (a, c, e). Data in Fig.4a, c, e are presented as mean values ± SEM. Exact P values are reported in the figure. Source data are provided as a Source Data file.