Journal: Virus Research

Article Title: Imidazonaphthyridine effects on Chikungunya virus replication: Antiviral activity by dependent and independent of interferon type 1 pathways

doi: 10.1016/j.virusres.2022.199029

Figure Lengend Snippet: RO8191 activity on CHKV -nanoluc replication. A) Representative structure of CHIKV- nanoluc genome [PDB: 6NK5]( Basore et al., 2019 ; Metz and Pijlman, 2016 ). B) Imidazonaphthyridine chemical structure (RO4948191 or RO8191) (CAS No.: 691,868–88–9; https://www.medchemexpress.com/ro8191.html ). C) Representative scheme of the infection assays. BHK-21 cells were treated with two-fold serial dilutions of RO8191 at concentrations ranging from 0.46 to 60 µM. CHIKV- nanoluc replication was quantified by measuring nanoluciferase activity (indicated by a black square) and cell viability using MTT assay (indicated by a gray circle). Mean ± SD values from a minimum of three independent experiments, each measured in triplicate, are represented. All images were generated using GraphPad Prism 8 and GIMP 2.1v.

Article Snippet: B) Imidazonaphthyridine chemical structure (RO4948191 or RO8191) (CAS No.: 691,868–88–9; https://www.medchemexpress.com/ro8191.html ).

Techniques: Activity Assay, Infection, MTT Assay, Generated

Journal: Virus Research

Article Title: Imidazonaphthyridine effects on Chikungunya virus replication: Antiviral activity by dependent and independent of interferon type 1 pathways

doi: 10.1016/j.virusres.2022.199029

Figure Lengend Snippet: RO8191 activity on different stages of CHIKV- nanoluc replicative cycle in BHK-21 cells. A) BHK-21 cells were treated with RO8191 for 1 h, cells were washed with PBS to remove the compound, and were infected with CHIKV- nanoluc at MOI 0.1 for 1 h. Then, the medium was removed, cells were washed to remove unbound virus, and fresh medium was added. B) BHK-21 cells were infected with CHIKV- nanoluc (MOI 0.1) and simultaneously treated with RO8191 for 1 h. Then, cells were washed to completely remove the inoculum, and a fresh medium was added. C) RO8191 and CHIKV- nanoluc at MOI 5 were incubated for 1 h (representative inoculum). Then, the inoculum was added to the cells for an additional hour. Cells were washed to remove the inoculum and fresh medium was added. D) BHK-21 cells were infected with CHIKV- nanoluc (MOI 0.1) for 1 h, cells were washed with PBS to remove unbound virus, and were treated with RO8191. All infection assays were quantified 16 h.p.i through the measurement of luminescence levels. Schematic representation of each time-based assay as indicated by BHK-21 cells (black bars), RO8191 (gray bars), CHIKV- nanoluc (orange bars), and CHIKV-RO8191 inoculum (blue tube). E) Schematic representation of the time-based BHK-CHIKV cell assay as indicated by the BHK-CHIKV-NCT cells (black bars) and RO8191 (gray bars). BHK-CHIKV-NCT cells were incubated for 72 h with the RO8191 compound at concentrations ranging from 60 to 0.46 µM. Then, cells were washed with PBS and lysed with Passive Lysis Buffer to measure the activity of Renilla . The reduction of Renilla activity in BHK-CHIKV-NCT cells is shown by 50% of the highest concentrations (EC 50 ) of RO8191 (black squares) and an effect in cell viability (CC 50 ) using MTT assay (gray circles). Mean ± SD values of a minimum of three independent experiments, each measured in triplicate. (***) P <0.001, (**) P <0.01, and (*) P <0.05 are represented. Images were generated using GraphPad Prism 8 and GIMP 2.1v.

Article Snippet: B) Imidazonaphthyridine chemical structure (RO4948191 or RO8191) (CAS No.: 691,868–88–9; https://www.medchemexpress.com/ro8191.html ).

Techniques: Activity Assay, Infection, Virus, Incubation, Lysis, MTT Assay, Generated

Journal: Virus Research

Article Title: Imidazonaphthyridine effects on Chikungunya virus replication: Antiviral activity by dependent and independent of interferon type 1 pathways

doi: 10.1016/j.virusres.2022.199029

Figure Lengend Snippet: BHK-21 cells express type-1 IFN (α/β) during RO8191 treatment. A) Percentage of CHIKV WT infection by measuring mRNA-CHIKV WT in supernatant of BHK-21 cells. B) CHIKV WT RNA titers by measuring mRNA-CHIKV WT in supernatant of BHK-21 cells. C) Percentage of CHIKV WT infection by measuring mRNA-CHIKV WT per mL in cell lysates of BHK-21 cells. D) CHIKV WT RNA titers by measuring mRNA-CHIKV WT per mL detected in cell lysates of BHK-21 cells. The CHIKV infection is presented in black bars while CHIKV in the presence of RO8191 treatment is demonstrated in gray bars. E) mRNA means of IFN-α expressed by BHK-21 during infection and/or treatment with RO8191. F) mRNA mean of IFN-β expressed by BHK-21 during infection and/or treatment with RO8191. BHK-21 cells were infected with CHIKV WT (MOI 0.1 PFU/well) in the presence or absence of RO8191 at 5 µM for 16 h. The supernatant and cell lysates were collected and processed to RNA extraction. cDNA was produced and mRNA levels was quantified by RT-qPCR. Mean values of three independent experiment each measured in triplicate, are represented. (****) P <0.0001. Images were generated using GraphPad Prism 8 and GIMP 2.1v.

Article Snippet: B) Imidazonaphthyridine chemical structure (RO4948191 or RO8191) (CAS No.: 691,868–88–9; https://www.medchemexpress.com/ro8191.html ).

Techniques: Infection, RNA Extraction, Produced, Quantitative RT-PCR, Generated

Journal: Virus Research

Article Title: Imidazonaphthyridine effects on Chikungunya virus replication: Antiviral activity by dependent and independent of interferon type 1 pathways

doi: 10.1016/j.virusres.2022.199029

Figure Lengend Snippet: RO8191 activity against CHIKV- nanoluc infection in Vero-E6 cells. A) Representative scheme of the infection assays. Vero-E6 cells were treated with three-fold serial dilutions of RO8191 at concentrations ranging from 0.02 to 60 µM. CHIKV replication was quantified by measuring nanoluciferase activity (indicated by a black square) and cell viability using an MTT assay (indicated by a gray circle). Values of EC 50 , CC 50, and SI were obtained from the treatment of Vero-E6 cells treated with RO8191. B) Vero-E6 cells were treated with RO8191 for 1 h, then the cells were washed with PBS and infected with CHIKV- nanoluc virus at MOI 0.1 for 1 h. The medium was removed, cells were washed with PBS and fresh medium was added. C) Vero-E6 cells were infected with CHIKV- nanoluc (MOI 0.1) and simultaneously treated with RO8191 for 1 h. Then, cells were washed, and fresh medium was added. D) RO891 and CHIKV- nanoluc at MOI 5 were incubated for 1 h, then the inoculum was added to cells for an extra 1 h, cells were washed, and fresh medium was added. E) Vero-E6 cells were infected with CHIKV- nanoluc (MOI 0.1) for 1 h, cells were washed with PBS and treated with RO8191. Schematic representation of each time-based assay as indicated by Vero-E6 cells (blue bars), RO8191 (gray bars), CHIKV- nanoluc (orange bars), and CHIKV- nanoluc /RO8191 inoculum (blue tube). Mean ± SD values of a minimum of three independent experiments, each measured in triplicate, are represented. (****) P <0.0001, (***) P <0.001 and (*) P <0.05. Images were generated using GraphPad Prism 8 and GIMP 2.1v.

Article Snippet: B) Imidazonaphthyridine chemical structure (RO4948191 or RO8191) (CAS No.: 691,868–88–9; https://www.medchemexpress.com/ro8191.html ).

Techniques: Activity Assay, Infection, MTT Assay, Virus, Incubation, Generated

Journal: Virus Research

Article Title: Imidazonaphthyridine effects on Chikungunya virus replication: Antiviral activity by dependent and independent of interferon type 1 pathways

doi: 10.1016/j.virusres.2022.199029

Figure Lengend Snippet: Localization and ChemPLP values showed between RO8191 and CHIKV proteins by molecular docking calculations.

Article Snippet: B) Imidazonaphthyridine chemical structure (RO4948191 or RO8191) (CAS No.: 691,868–88–9; https://www.medchemexpress.com/ro8191.html ).

Techniques:

Journal: Virus Research

Article Title: Imidazonaphthyridine effects on Chikungunya virus replication: Antiviral activity by dependent and independent of interferon type 1 pathways

doi: 10.1016/j.virusres.2022.199029

Figure Lengend Snippet: 2D and 3D interactions between RO8191 and CHIKV proteins showed by molecular docking analysis. A) RO8181 interacts with CHIKV glycoproteins mainly through Hydrogen Bond (green lines) and alkyl ligands (pink lines). B) RO8191 showed interactions with nsP1 protein mainly through Conventional Hydrogen Bonds (green circles) interactions. C) RO8191 interacts with nsP2 mainly through Alkyl (pink lines) and Hydrogen Bonds (green lines) interactions. D) RO8191 interacts with nsP3 mainly through Hydrogens Bonds ligands and interactions (green lines and circles). E) RO8191 interactions with nsP4 mainly through Conventional Hydrogens Bonds (green lines) and Halogen (blue lines) ligands. RO8191 (PDB: 3R0L) and the CHIKV proteins (glycoprotein complex [PDBid: 3N42], nsP1 [PDBid: 7DOP], nsP2 [PDBid:4ZTB], nsP3 [PDBid:6W8Z], and database representative sequence of the nsP4 extracted from the virus polyprotein [uniport-id: Q8JUX6]) were docked employing the GOLD program site using a genetic algorithm (GA) and the score ChemPL. Images were generated using GraphPad Prism 8 and GIMP 2.1v.

Article Snippet: B) Imidazonaphthyridine chemical structure (RO4948191 or RO8191) (CAS No.: 691,868–88–9; https://www.medchemexpress.com/ro8191.html ).

Techniques: Sequencing, Virus, Generated

Journal: Virus Research

Article Title: Imidazonaphthyridine effects on Chikungunya virus replication: Antiviral activity by dependent and independent of interferon type 1 pathways

doi: 10.1016/j.virusres.2022.199029

Figure Lengend Snippet: Infrared spectroscopy indicates molecular interactions between CHIKV virions with RO8191. A) Representative scheme of the ATR-FTIR technology with CHIKV virions (black line), RO8191 (blue line), and CHIKV virion plus incubation with RO8191 (red line). B) The representative infrared average spectrum of normalized mean spectra of CHIKV virion (black line), RO8191 (blue line), and CHIKV virion plus RO8191 (red line) employing ATR-FTIR platform from 1800 to 800 cm −1 . C-F) The representative mean spectra of second derivative analysis to the vibrational modes at 1496 cm −1 ( C ), 1465 cm −1 ( D ), 1448 cm −1 ( E ), 1333 cm −1 ( F ), and 1078 cm −1 ( G ). Images were generated using GraphPad Prism 8 and GIMP 2.1v.

Article Snippet: B) Imidazonaphthyridine chemical structure (RO4948191 or RO8191) (CAS No.: 691,868–88–9; https://www.medchemexpress.com/ro8191.html ).

Techniques: Spectroscopy, Incubation, Generated

Journal: Advanced Science

Article Title: hESC‐Derived Epicardial Cells Promote Repair of Infarcted Hearts in Mouse and Swine

doi: 10.1002/advs.202300470

Figure Lengend Snippet: The IFN‐I receptor agonist RO8191 attenuates the anti‐inflammatory and cardiac reparative effects of hEPs. A) Cardiac performance measured by echocardiography. All mice received intragastric administration of 10 mg kg −1 RO8191 or vehicle control (corn oil) 30 min before MI surgery. n = 6, 8, 6, and 8 in the MI, MI + hEPs, MI + RO8191, and MI + hEPs + RO8191 groups. ** p < 0.01, *** p < 0.001, and n.s.(no significant difference) versus the MI group. # p < 0.05, ## p < 0.01, and n.s. as indicated. B) Representative images and quantitative analysis of scar sizes stained with Masson's trichrome at day 28 post‐MI. n = 6 to 7. Scale bar: 1 mm. C,D) Representative images and quantitative data of immunofluorescent staining for CD31 + endothelial cells (C, n = 5 to 6) and LYVE1 + lymphatic vessels (D, n = 4 to 7) in the border zones at day 28 post‐MI. Scale bar: 50 µm. E) Representative images and quantitative data of immunofluorescent staining for TUNEL + CMs in the border zones at day 3 post‐MI. n = 4 to 5. Scale bar: 20 µm. F) Representative images and quantitative data of immunofluorescent staining for CD68 + cells and CD68 + CD206 + macrophages in the border zones at day 3 post‐MI. n = 5 to 6. Scale bar: 50 µm. Data are means ± S.E.M. Two‐way ANOVA followed by Bonferroni's multiple analysis in A to F. * p < 0.05, ** p < 0.01, and *** p < 0.001. n.s.: no significant difference.

Article Snippet: RO8191 (APExBIO) was diluted with corn oil and intragastrically administrated at 10 mg kg −1 within 30 min before MI injury.

Techniques: Control, Staining, TUNEL Assay