Journal: Viruses

Article Title: Molnupiravir and Its Active Form, EIDD-1931, Show Potent Antiviral Activity against Enterovirus Infections In Vitro and In Vivo

doi: 10.3390/v14061142

Figure Lengend Snippet: The molecular formula of EIDD−1931 and EIDD−2801 and the in vitro antiviral effects against EV−A71. ( A ) The molecular formula of EIDD−1931 and EIDD−2801. ( B – G ) The antiviral activities of EIDD−1931 ( B – D ) and EIDD2801 ( E – G ) against EV−A71 in different cells lines. RD cells, Vero cells, and Huh7 cells were infected with the EV−A71 H strain at 100 × TCID 50 . Different doses of the test compounds were then added. At 72 h.p.i, the antiviral parameters were measured. The antiviral effects and cytotoxicity of EIDD−1931 and EIDD−2801 were measured using a CellTiter−Glo cell viability assay kit. The EC 50 and CC 50 were calculated using Origin 9.0 software. SI = CC 50 /IC 50 .

Article Snippet: EIDD-1931(cat no. T8498) and EIDD-2801 (cat no. T8309) powders were purchased from TargetMol (Shanghai, China), and NITD008 (7-Deaza-2′-C-acetylene-adenosine, cat no. HY-12957) powders were purchased from MCE (Shanghai, China).

Techniques: In Vitro, Infection, Viability Assay, Software

Journal: Viruses

Article Title: Molnupiravir and Its Active Form, EIDD-1931, Show Potent Antiviral Activity against Enterovirus Infections In Vitro and In Vivo

doi: 10.3390/v14061142

Figure Lengend Snippet: Antiviral activities of EIDD−1931 and EIDD−2801 against EV−A71 in vitro. ( A – D ) EV−A71 virus particle yields and viral RNA reduction assay. RD cells were infected with EV71 at an MOI of 0.1 PFU/mL in the presence of different concentrations of EIDD−1931 or EIDD−2801. After 30 h, the virus particle yields in the supernatant were measured by TCID 50 . Total cellular RNA was extracted and subjected to qRT−PCR to measure viral RNA expression. All data are shown as means ± standard deviation from three independent experiments. Statistical significance was calculated using one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. ( E – G ) Inhibitory effects of EIDD-1931 and EIDD−2801 on viral VP1 protein. ( E ) RD cells were infected with EV−A71 at an MOI of 1 PFU/mL in the presence of EIDD−1931 or EIDD−2801. At 16 h.p.i., the cells were fixed for immunofluorescence assays. EV−A71 VP1 proteins were stained using mouse anti−EV−A71 antibody (12D7) (red), and cell nuclei were stained using Hoechst 33,342 (blue), scale bar: 50 µm. ( F , G ) RD cells infected with EV−A71 (MOI = 1) were treated with the test compounds at the indicated concentrations. At 24 h post infection, the cells were collected for Western blot analysis using anti−EV−A71VP1 antibodies (12D7). ( H , I ) Proteins levels of EV−A71 VP1 were quantified by ImageJ software. * p < 0.05, **** p < 0.0001.

Article Snippet: EIDD-1931(cat no. T8498) and EIDD-2801 (cat no. T8309) powders were purchased from TargetMol (Shanghai, China), and NITD008 (7-Deaza-2′-C-acetylene-adenosine, cat no. HY-12957) powders were purchased from MCE (Shanghai, China).

Techniques: In Vitro, Infection, Quantitative RT-PCR, RNA Expression, Standard Deviation, Immunofluorescence, Staining, Western Blot, Software

Journal: Viruses

Article Title: Molnupiravir and Its Active Form, EIDD-1931, Show Potent Antiviral Activity against Enterovirus Infections In Vitro and In Vivo

doi: 10.3390/v14061142

Figure Lengend Snippet: EIDD-1931 and EIDD-2801 exert in vivo antiviral effects against EV-A71 infection. Four groups of 1-day-old ICR suckling mice were i.p. infected with 10 6 PFU of EV-A71 virus (H strain), followed by i.p. treatment with EIDD-1931, EIDD-2801, or vehicle at the indicated dosages for 7 consecutive days. Survival ( A , C ) and body weight ( B , D ) were recorded every day until 21 d.p.i. Data for B and D are presented as means ± standard deviations. Survival data were analyzed with a log-rank test. ** p < 0.01, **** p < 0.0001.

Article Snippet: EIDD-1931(cat no. T8498) and EIDD-2801 (cat no. T8309) powders were purchased from TargetMol (Shanghai, China), and NITD008 (7-Deaza-2′-C-acetylene-adenosine, cat no. HY-12957) powders were purchased from MCE (Shanghai, China).

Techniques: In Vivo, Infection

Journal: Viruses

Article Title: Molnupiravir and Its Active Form, EIDD-1931, Show Potent Antiviral Activity against Enterovirus Infections In Vitro and In Vivo

doi: 10.3390/v14061142

Figure Lengend Snippet: EIDD-1931 and EIDD-2801 reduce the viral loads in various tissues of 1-day-old ICR suckling mice. Three groups of 1-day-old ICR suckling mice were i.p. infected with 10 6 PFU of EV-A71 virus (H strain), followed by i.p. treatment with EIDD-1931 and EIDD-2801 at a dosage of 200 mg/kg for 4 consecutive days. Then, the mice were euthanized, and brain ( A ), heart ( B ), intestine ( C ), liver ( D ), limb muscles ( E ) and lung ( F ) were separately harvested to determine the viral RNA loads using qRT-PCR. Data are presented as means ± standard deviations, and Student’s unpaired t test was performed for statistical analysis. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: EIDD-1931(cat no. T8498) and EIDD-2801 (cat no. T8309) powders were purchased from TargetMol (Shanghai, China), and NITD008 (7-Deaza-2′-C-acetylene-adenosine, cat no. HY-12957) powders were purchased from MCE (Shanghai, China).

Techniques: Infection, Quantitative RT-PCR

Journal: Communications Biology

Article Title: PIKfyve-specific inhibitors restrict replication of multiple coronaviruses in vitro but not in a murine model of COVID-19

doi: 10.1038/s42003-022-03766-2

Figure Lengend Snippet: a Groups of mice were treated intraperitoneally with PIKfyve inhibitors WX8 or NDF once daily beginning 1 day pre-intranasal-challenge with 1 × 10 5 plaque forming units SARS-CoV-2 (B.1.351). EIDD-2801 dosed twice a day was used as a positive treatment control and uninfected treatment controls were included to assess cytotoxicity. Image created using BioRender. b Weight changes were determined for 4 days post-challenge, plotted as the group mean with error bars indicating the ±SD. c Infectious viral loads from lung homogenates at 2 (black) or 4 (gray) days post SARS-CoV-2 challenge. d Lungs were collected at 2- or 4-days post-challenge and stained with hematoxylin and eosin to assess bronchiolar and alveolar damage and immune cell infiltration (500-μm scale bar shown at bottom left, representative for all panels). Mixed-effects analysis was used to compare differences in weight change or viral loads from lung homogenates between infection treatment groups and the vehicle-treated control group; ** p ≤ 0.01, **** p ≤ 0.0001. dpi, days post-infection; PO, oral dosing; IP, intraperitoneal; IN, intranasal.

Article Snippet: EIDD-2801 (150 mg/kg, WuXi AppTec) was dosed orally, twice daily, starting one day prior to infection was used as a positive control for all experiments, as it has been shown to be efficacious in animal models previously , .

Techniques: Staining, Infection

Journal: Communications Biology

Article Title: PIKfyve-specific inhibitors restrict replication of multiple coronaviruses in vitro but not in a murine model of COVID-19

doi: 10.1038/s42003-022-03766-2

Figure Lengend Snippet: a Groups of mice were treated intraperitoneally with PIKfyve inhibitors Apilimod, WX8, or NDF once daily beginning 1 day post-intranasal-challenge with 1 × 10 3 plaque forming units SARS-CoV-2 (MA-10). EIDD-2801 dosed twice a day was used as a positive treatment control and uninfected treatment controls were included to assess cytotoxicity. Image created using Biorender. b Weight changes were determined for 4 days post-challenge, plotted as the group mean with error bars indicating the ±SD. c Infectious viral loads from lung homogenates at 2- (black) or 4- (gray) days post SARS-CoV-2 challenge. d Survival curves for treatment groups. e Lungs were collected at 2- or 4-days post-challenge and stained with hematoxylin and eosin to assess bronchiolar and alveolar damage and immune cell infiltration (500-μm scale bar shown at bottom left, representative for all panels). Mixed-effects analysis was used to compare differences in weight change or viral loads from lung homogenates between infection treatment groups and the vehicle-treated control group; * p ≤ 0.1, *** p ≤ 0.001, **** p ≤ 0.0001. dpi, days post-infection; PO, oral dosing; IP, intraperitoneal; IN, intranasal.

Article Snippet: EIDD-2801 (150 mg/kg, WuXi AppTec) was dosed orally, twice daily, starting one day prior to infection was used as a positive control for all experiments, as it has been shown to be efficacious in animal models previously , .

Techniques: Staining, Infection

Journal: Frontiers in Pharmacology

Article Title: Preclinical Pharmacokinetics and In Vitro Properties of GS-441524, a Potential Oral Drug Candidate for COVID-19 Treatment

doi: 10.3389/fphar.2022.918083

Figure Lengend Snippet: Anti-SARS-CoV-2 activity of GS-441524 and Remdesivir. (A) GS-441524 inhibited SARS-CoV-2 CPE in Vero E6 cells with an EC 50 of 1.86 μM compared to remdesivir (EC 50 of 7.43 μM). Neither compounds showed cytotoxicity at the concentrations used. (B) GS-441524 reduced viral load of SARS-CoV-2 infection in the 3D EpiAirway HAE tissue model. TCID50/mL was determined for remdesivir, GS-441524 and EIDD-2801 after (B) 24 h or (C) 96 h of SARS-CoV-2 infection. All graphs show mean ± standard error of the mean (SEM).

Article Snippet: For Vero E6 CPE assay and EpiAirway efficacy study, GS-441524 and remdesivir were purchased from MedChem Express (Monmouth Junction, NJ), and EIDD-2801 was purchased from Cayman Chemical (Ann Arbor, MI).

Techniques: Activity Assay, Infection