ledipasvir  (MedChemExpress)

Journal: PLoS ONE

Article Title: The combination of the NS5A and cyclophilin inhibitors results in an additive anti-HCV inhibition in humanized mice without development of resistance

doi: 10.1371/journal.pone.0251934

Figure Lengend Snippet: Humanized-liver mouse model for HCV infection. A. Experimental design for the creation and validation of the HCV infection model in humanized-liver chimeric MUP-uPA-SCID/Beige mice. B. Chemical structures of the NS5Ai Velpatasvir and Ledipasvir, and the CypI CRV431 and Alisporivir.

Article Snippet: DrugsCRV431 was manufactured by chemical modification of cyclosporin A. Alisporivir, Velpatasvir, Ledipasvir and Sofosbuvir were purchased from MedChemExpress.

Techniques: Infection, Mouse Assay

Journal: PLoS ONE

Article Title: The combination of the NS5A and cyclophilin inhibitors results in an additive anti-HCV inhibition in humanized mice without development of resistance

doi: 10.1371/journal.pone.0251934

Figure Lengend Snippet: Individual analysis of anti-HCV efficacy of Velpatasvir, Ledipasvir, CRV431 and Alisporivir in humanized-liver chimeric mice. MUP-uPA-SCID-Beige mice implanted with human hepatocytes were infected i.v. with plasma from HCV-infected chimpanzee (100 infectious doses (CID 50 )/mL of GT1a, GT2a, GT3a and GT4a). Drugs (50 mg/kg) were given orally 9 weeks post-infection. Blood was collected retro-orbitally every week until week 15 post-HCV infection. Ten mice per treatment. Viral replication (HCV RNA copies/ml of serum) was quantified by real-time reverse transcription PCR. Error bars corresponds to the standard deviation (SD) between 10 animals. At week 12 and 15 post-infection, SD are small due to the minimal viral loads due to the drug treatment efficacy. Data are representative of two independent experiments.

Article Snippet: DrugsCRV431 was manufactured by chemical modification of cyclosporin A. Alisporivir, Velpatasvir, Ledipasvir and Sofosbuvir were purchased from MedChemExpress.

Techniques: Mouse Assay, Infection, Polymerase Chain Reaction, Standard Deviation

Journal: PLoS ONE

Article Title: The combination of the NS5A and cyclophilin inhibitors results in an additive anti-HCV inhibition in humanized mice without development of resistance

doi: 10.1371/journal.pone.0251934

Figure Lengend Snippet: HCV rebound analysis in humanized-liver chimeric mice. MUP-uPA-SCID-Beige mice implanted with human hepatocytes were infected i.v. with plasma from HCV-infected chimpanzee (100 infectious doses (CID 50 )/mL of GT1a, GT2a, GT3a and GT4a). Drugs—Velpatasvir, Ledipasvir, CRV431, Alisporivir—(50 mg/kg) were orally given alone or in combination 2 months post-infection. Ten mice per treatment. Blood was collected until month 9 post-HCV infection. Viral replication (HCV RNA copies/ml of serum) was quantified by real-time reverse transcription PCR. At week 12 and 15 post-infection, SD are small due to the minimal viral loads due to the drug treatment efficacy. Data are representative of two independent experiments.

Article Snippet: DrugsCRV431 was manufactured by chemical modification of cyclosporin A. Alisporivir, Velpatasvir, Ledipasvir and Sofosbuvir were purchased from MedChemExpress.

Techniques: Mouse Assay, Infection, Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: The combination of the NS5A and cyclophilin inhibitors results in an additive anti-HCV inhibition in humanized mice without development of resistance

doi: 10.1371/journal.pone.0251934

Figure Lengend Snippet: Combinational analysis of anti-HCV efficacy of Velpatasvir, Ledipasvir, Alisporivir and CRV431 in humanized-liver chimeric mice. A. MUP-uPA-SCID-Beige mice implanted with human hepatocytes were infected i.v. with plasma from HCV-infected chimpanzee (100 infectious doses (CID 50 )/mL of GT1a, GT2a, GT3a and GT4a). Drugs were orally given alone (50 mg/kg) or in combination (50/50 mg/kg) 9 weeks post-infection. Blood was collected until week 15 post-HCV infection. Ten mice per treatment. Viral replication (HCV RNA copies/ml of serum) was quantified by real-time reverse transcription PCR. Error bars corresponds to the SD between 10 animals. At week 12 and 15 post-infection, SD are small due to the minimal viral loads due to the drug treatment efficacy. B. Statistical analyses for drug combination effect between HCV genotypes. C. Statistical analyses for Ledipasvir combination effect on HCV GT1a, GT2a, GT3a and GT4a. D. Statistical analyses for Velpatasvir combination effect on HCV GT1a, GT2a, GT3a and GT4a. For each treatment/genotype combination, we simulated 1,000 draws using the mean and standard deviation from the experimental results to account for the uncertainty in estimating the Area Under the Curve for the level of viremia. Area Under the Curve was estimated using the trapezoidal method. Groups were compared using t-tests and the permutation test. P-values were adjusted for multiple comparisons using the Bonferroni correction. Data are representative of two independent experiments.

Article Snippet: DrugsCRV431 was manufactured by chemical modification of cyclosporin A. Alisporivir, Velpatasvir, Ledipasvir and Sofosbuvir were purchased from MedChemExpress.

Techniques: Mouse Assay, Infection, Polymerase Chain Reaction, Standard Deviation

Journal: Viruses

Article Title: Visualization of Positive and Negative Sense Viral RNA for Probing the Mechanism of Direct-Acting Antivirals against Hepatitis C Virus

doi: 10.3390/v11111039

Figure Lengend Snippet: Early impact of different classes of HCV inhibitors on HCV RNA and NS5A. ( A ) Huh-7.5.1 cells were infected with HCV Jc1/Gluc2A at an MOI of 0.5. After 48 h, the cells were treated with various inhibitors (Danoprevir, 0.32 μM; Ledipasvir, 3 μM; Daclatasvir, 3.2 nM; Sofosbuvir, 20 μM), then after a further 8 h cells were fixed and probed sequentially for (−)RNA, (+)RNA and NS5A. Finally, nuclei were stained with DAPI. Cells were imaged on a Leica SP8 confocal microscope using a 63× oil-immersion objective. ( B ) Representative images from each inhibitor showing (−)RNA in green, (+)RNA in red, NS5A in gray and nuclei in blue. Boxed regions shown enlarged. Scale bars represent 10 µm. Early impact of different classes of HCV inhibitors on HCV RNA and NS5A. (C and D) Fields of view were captured as described in Figure 3 , then analyzed using Gen5 software (BioTek). Abundance of (+) and (−)RNA strands was quantified by: ( C ) number of RNA foci per cell; ( D ) RNA fluorescence intensity per cell in the field of view; and ( E ) RT-qPCR to determine copy numbers of the of (+) and (−)RNA using specific primers. All values are expressed as a percentage of the DMSO-treated sample. Error bars indicate standard error of the mean for three independent experiments. Significance of differences between DMSO and drug treatments was calculated using one-way ANOVA and Tukey’s post-test for multiple comparisons. Differences were not significant unless otherwise indicated. *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001. P values shown on columns indicate comparison to DMSO. Lines indicate comparisons between test samples. Lines group samples, and are color coordinated for (+)RNA and (−)RNA. Flat lines group all samples beneath them, inverted Vs indicate comparison of the samples under the ends of the lines. DNV, Danoprevir; LDV, Ledipasvir; DCV, Daclatasvir; SOF, Sofosbuvir. ( F ) Comparison of intracellular HCV transcript number under control (DMSO) conditions as determined by strand-specific RT-qPCR and bDNA FISH. Error bars indicate standard error of the mean; no additional statistical analyses were performed.

Article Snippet: Ledipasvir (LDV, GS-5885) was purchased from MedChem Express.

Techniques: Infection, Staining, Microscopy, Software, Fluorescence, Quantitative RT-PCR, Fluorescence In Situ Hybridization

Journal: bioRxiv

Article Title: A visualizable hepatitis A virus and hepatitis C virus coinfection model in vitro: coexistence of two hepatic viruses under limited competition in viral RNA synthesis

doi: 10.1101/709071

Figure Lengend Snippet: Extra-added rNTPs increases both HAV and HCV replication in coinfection. rNTPs and other drugs were added at the time of four hours post HAV(HM175/18f) infection. The working concentrations of rNTP, IFN-α, Simeprevir, and Ledipasvir were 0.1mM, 1000IU/ml, 200nM and 200nM respectively; Sofosbuvir concentration (nM) was used as indicated. Mock represents 0.2% DMSO. 48 hours after HAV infection, cells were washed once with 1×PBS and harvested. RT-qPCR was used to quantify the intracellular viral RNA amounts and relative change was created by dividing the mock. (A) Effect of rNTPs on HAV replication and (B) Effect of rNTPs on HCV replication. (C) Effect of sofosbuvir on HAV replication and (D) Effect of sofosbuvir on HAV replication. (E) Effect of indicated drugs on HAV replication. (F) Effect of indicated drugs on HCV replication. Data represent the mean of three independent assays.

Article Snippet: Sofosbuvir, Simeprevir, and Ledipasvir were purchased from MedChem Express.

Techniques: Infection, Concentration Assay, Quantitative RT-PCR