molnupiravir  (Shionogi)

Journal: Cell Reports Medicine

Article Title: Human liver-derived organoids recapitulate Oropouche virus infection and manifestation, enabling antiviral drug discovery

doi: 10.1016/j.xcrm.2026.102646

Figure Lengend Snippet: Antiviral drug profiling identified molnupiravir as a potent inhibitor of OROV infection (A) Schematic diagram of profiling RdRp inhibitors, prioritizing clinically used drugs, in OROV-infected liver organoids. (B) Fold changes of OROV RNA levels in infected organoids after 48-h treatment with the selected RdRp inhibitors. (C–E) (C) Potential docking pocket of RdRp protein predicted by CurPocket. RNA-protein contact active site cavity is indicated as a dotted circle (D and E). Dose-response curves of molnupiravir (D) and NHC (E) in infected organoids, showing the 50% inhibitory concentration (IC 50 ) and 50% cytotoxic concentration (CC 50 ) values ( n = 4 biological replicates). (F) Intracellular OROV RNA levels ( n = 5 biological replicates) and corresponding cell viability ( n = 8 biological replicates) following 48- and 96-h treatment with NHC (0.1 or 1 μM). (G) Infectious viral titers in culture supernatant from organoids treated with NHC at 48 and 96 h ( n = 4 biological replicates). (H) Schematic of the time-of-addition experiment to assess the timing of NHC-mediated antiviral activity. (I) OROV RNA levels under virucidal treatment (virucidal, n = 7 biological replicates), pre-treatment (pre, n = 6 biological replicates), treatment during virus inoculation (during, n = 7 biological replicates), and post-treatment conditions (post, n = 5 biological replicates). Data are shown as relative levels compared to matched untreated controls in each group. The post-treatment group is same as the condition of 48-h treatment with 1 μM concentration in (F). (J) OROV RNA levels in the supernatant of cultured organoids without or with NHC treatment (1 or 5 μM) over a 15-day period. Data are shown as relative levels normalized to untreated controls ( n = 5 biological replicates). Statistical comparisons between treatment groups were performed using two-tailed Mann-Whitney U tests (F, G, and I). Statistical analysis for longitudinal data were performed using mixed-effects model (REML) (J). Data represent mean ± SEM; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001.

Article Snippet: Molnupiravir (EIDD-2801) , MedChemExpress , Cat#HY-135853.

Techniques: Infection, Concentration Assay, Activity Assay, Virus, Cell Culture, Two Tailed Test, MANN-WHITNEY

Journal: Cell Reports Medicine

Article Title: Human liver-derived organoids recapitulate Oropouche virus infection and manifestation, enabling antiviral drug discovery

doi: 10.1016/j.xcrm.2026.102646

Figure Lengend Snippet: Molnupiravir attenuated virus-induced cell death and modulates host antiviral responses in OROV-infected liver organoids (A) Representative images of OROV-2024-infected organoids without (top) or with (bottom) 96-h NHC (1 μM) treatment by immunostaining with the antibody against OROV Gc glycoprotein (red), phalloidin for F-actin (green), and DAPI for nuclei (blue). Scale bars: 50 , 25, and 10 μm. (B) Representative images of OROV-2024 infected organoids without (top) or with (bottom) 96-h NHC (1 μM) treatment by immunostaining with the antibody against OROV Gc glycoprotein (red), cleaved-caspase-3 (green), and nuclei (DAPI, blue). Scale bars: 35 and 10 μm. (C) LDH release in culture supernatant of organoids at 48 and 96 h post-inoculation, comparing uninfected, OROV-infected, and NHC-treated groups (1 μM) ( n = 4 biological replicates). (D) GSEA in OROV-infected organoids with NHC treatment for 96 h compared to organoids with OROV infection alone. (E) Heatmap of the top 50 significantly regulated genes comparing uninfected controls, OROV-infected organoids at 96 h post-inoculation, and OROV-infected organoids with 96-h NHC treatment. (F) Heatmap of selected ISGs comparing OROV-infected organoids at 96 h post-inoculation and OROV-infected organoids with 96-h NHC treatment. ISGs were sourced from a published large-scale ISG screen and mapped to our RNA-seq dataset. The color scale represents row-wise Z score of gene expression across samples. Data are representative of four biological replicates from one RNA-seq experiment (D–F). Data represent mean ± SEM; statistical analysis by Mann-Whitney U test; ∗ p < 0.05; ns, not significant.

Article Snippet: Molnupiravir (EIDD-2801) , MedChemExpress , Cat#HY-135853.

Techniques: Virus, Infection, Immunostaining, RNA Sequencing, Gene Expression, MANN-WHITNEY

Journal: Cell Reports Medicine

Article Title: Human liver-derived organoids recapitulate Oropouche virus infection and manifestation, enabling antiviral drug discovery

doi: 10.1016/j.xcrm.2026.102646

Figure Lengend Snippet: Harnessing host interferon response for developing molnupiravir-based antiviral combination (A) Schematic illustration of serial passaging with ( n = 3 biological replicates) or without ( n = 3 biological replicates from one passaging experiment series) NHC treatment of escalating concentrations in Vero cells. (B) Identification of mutations by whole-genome sequencing of OROV using Nanopore sequencing technology. (C) Sankey-bubble plot showing significantly enriched immune and interferon-related pathways in OROV-infected human adult liver-derived organoids at 96 h post-inoculation. Data are representative of four biological replicates from one RNA-seq experiment. (D) Quantification of OROV RNA levels in organoids with or without JAK inhibitor treatment ( n = 5 biological replicates). (E) Infectious viral titers in culture medium from organoids treated with or without JAK inhibitor ( n = 5 biological replicates). (F) Relative expression of selected ISGs in OROV-infected organoids treated with or without 10 μM JAK inhibitor ( n = 4 biological replicates). (G) Intracellular OROV RNA levels and corresponding cell viability following IFN-α treatment for 48 h with increasing concentrations (0, 1, 10, 100, 1,000 IU/mL) ( n = 4 biological replicates). (H) Infectious viral titers in culture medium following IFN-α treatment ( n = 4 biological replicates). (I) Heatmap showing induced expression of representative ISGs after 48-h treatment of IFN-α at different concentrations. Values represent log 2 (fold change) relative to untreated controls. (J and K) Dose-response matrix (J) and ZIP synergy landscape (K) of IFN-α and NHC combination treatment in OROV-infected organoids, generated by SynergyFinder 3.0. Data represent mean ± SEM; statistical analysis by two-tailed Mann-Whitney U test unless otherwise indicated; ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Molnupiravir (EIDD-2801) , MedChemExpress , Cat#HY-135853.

Techniques: Passaging, Sequencing, Nanopore Sequencing, Infection, Derivative Assay, RNA Sequencing, Expressing, Generated, Two Tailed Test, MANN-WHITNEY