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: Viruses

Article Title: Azelnidipine Exhibits In Vitro and In Vivo Antiviral Effects against Flavivirus Infections by Targeting the Viral RdRp

doi: 10.3390/v14061228

Figure Lengend Snippet: Enzymatic inhibition evaluation of ALP against flavivirus RdRp protein. ( A ) Real-time fluorescence changes in DENV RdRp enzymatic system inhibited by different concentrations of ALP (50–3.125 μΜ). ( B ) Dose–response inhibitory curve for ALP’s activity against the DENV RdRp protein. The nucleotide 3′-dATP was used as the positive control. ( C ) Surface plasmon resonance assay was conducted to determine the direct interactions between ALP and DENV RdRp protein. Immobilized DENV RdRp protein was coated onto a CM5 chip and incubated with gradient-diluted ALP for response unit monitoring. Representative results are as shown.

Article Snippet: Azelnidipine (MCE, Monmouth Junction, NJ, USA, cat #HY-B0023), 7-deaza-2′-C-acetylene-adenosine (NITD008; MCE, Monmouth Junction, NJ, USA, cat #HY-12957), 2′-C-methyladenosine (2′-CMA; TargetMol, Boston, MA, USA, cat #T16325), and 3′-dATP (Sigma, St. Louis, MO, USA, cat # C9137) were dissolved in DMSO and stored at −20 °C until used.

Techniques: Inhibition, Fluorescence, Activity Assay, Positive Control, SPR Assay, Incubation

Journal: Frontiers in Pharmacology

Article Title: The different effects of four adenosine receptors in liver fibrosis

doi: 10.3389/fphar.2024.1424624

Figure Lengend Snippet: NECA inhibited HSC activation and alleviated liver fibrosis. (A) Flowchart of animal experiments. (B,C) Serum ALT and AST levels in control, CCl 4 -induced mice, and NECA-treated mice. (D–F) Representative liver tissue sections of control, CCl 4 -, and NECA-treated mice were detected by HE, Masson, and Sirius Red staining. Scale bar: 50 μm; magnification: ×20. (G–J) qRT-PCR and IHC analysis showed the expression levels of α-SMA and Col1α1 in control, CCl 4 -, and NECA-treated mice. Scale bar: 50 μm; magnification: ×20. The positive area statistics of Masson and Sirius Red staining were measured using ImageJ software. (K,L) qRT-PCR analysis showed the expression levels of α-SMA and Col1α1 in LX2 cells. (M) Effect of NECA treatment on LX2 cell proliferation. (N) Expression of A1R, A2AR, A2BR, and A3R in LX2 cells with or without NECA treatment. * p < 0.05, ** p < 0.01, *** p < 0.0005, and **** p < 0.0001; ns: no significance.

Article Snippet: The mice were randomly divided into different groups: control group—olive oil (MACKLIN, China, Cat: 8001-25-0), 1 mL/kg i.p.; CCl 4- induced group [25% CCl 4 (MACKLIN, China, Cat: 56-23-5) (CCl 4 : olive oil = 1:3), 1 mL/kg i.p.]; NECA treatment group [1 mL/kg i.p. 25% CCl 4 plus 0.1 mg/kg i.p. NECA (GlpBio, Montclair, CA, United States, Cat: GC15304)]; A1R agonist group [1 mL/kg i.p. 25% CCl 4 plus 0.5 mg/kg i.p. CCPA (GlpBio, Montclair, CA, United States, Cat: GC45773)]; A2AR agonist group [1 mL/kg i.p. 25% CCl 4 plus 1 mg/kg i.p. CGS21680 (GlpBio, Montclair, CA, United States, Cat: GC10172)]; A2BR agonist group [1 mL/kg i.p. 25% CCl 4 plus 4 mg/kg i.p. BAY-606583 (TargetMol, China, Cat: 910487-58-0)]; and A3R agonist group [1 mL/kg i.p. 25% CCl 4 plus 200 μg/kg i.p. namodenoson (TargetMol, China, Cat: 163042-96-4)].

Techniques: Activation Assay, Control, Staining, Quantitative RT-PCR, Expressing, Software