1. Volume 68, Issue 3, Pages 412-420 (March 2018)
A novel orally available small molecule that inhibits hepatitis B virus expression 
Henrik Mueller, Steffen Wildum, Souphalone Luangsay, Johanna Walther, Anaïs Lopez, Philipp Tropberger, Giorgio Ottaviani, Wenzhe Lu, Neil John Parrott, Jitao David Zhang, Roland Schmucki, Tomas Racek, Jean-Christophe Hoflack, Erich Kueng, Floriane Point, Xue Zhou, Guido Steiner, Marc Lütgehetmann, Gianna Rapp, Tassilo Volz, Maura Dandri, Song Yang, John A.T. Young, Hassan Javanbakht 
Journal of Hepatology 
Volume 68, Issue 3, Pages (March 2018)
DOI: /j.jhep
Copyright © 2017 European Association for the Study of the Liver Terms and Conditions

2. Journal of Hepatology 2018 68, 412-420DOI: (10. 1016/j. jhep. 2017. 10
Copyright © 2017 European Association for the Study of the Liver Terms and Conditions

3. Fig. 1
Antiviral activities of RG7834, RO0321 and ETV in HBV-infected dHepaRG cells. Shown are the chemical structures of the test compounds, and mean dose-response curves of HBV DNA (blue line and squares), HBsAg (red line and circles) and HBeAg (black line and diamonds) inhibition (error bars represent standard deviations from at least four independent experiments). Mean EC50 and CC50 values ± standard deviation from at least four independent experiments are summarized in the table. ETV, entecavir; HBV, hepatitis B virus; HBeAg, hepatitis B e antigen; HBsAg, hepatitis B surface antigen; n.d., not determined; SD, standard deviation.
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2017 European Association for the Study of the Liver Terms and Conditions

4. Fig. 2
RNA deep sequencing analysis of the effect of RG7834 on HBV RNA transcripts and on host gene expression in HBV-infected dHepaRG cells. (A) Viral RNA transcript levels in the presence and absence of RG7834 (100 nM) at different time points of post-treatment initiation (error bars represent standard deviations). (B) HBV transcript-level quantification by nucleotide mapping in the presence and absence of RG7834 (100 nM) after 6 h (upper panel) and 24 h (lower panel) of treatment. Localization of HBV RNA transcripts mapped to the genome is shown above. (C) Host gene expression quantification in the presence of RG7834 after 6 h, 9 h, 12 h and 24 h compared to non-treated samples. Fold changes are shown as log2 scale. (D) Heatmap of expression of genes that are preferentially expressed in liver compared with other human organs and tissues. These ‘liver-enriched genes’ were identified from an integrative analysis of multiple large-scale transcriptomic datasets in our previous study.37 Mean expression of liver-enriched genes in three biological replicates are visualized with heatmap using units of log2-transformed Fragments Per Kilobase Million (FPKM). Each row represents one liver-enriched gene and each column one condition, i.e. 6 h, 9 h, 12 h, or 24 h after vehicle or RG7834 treatment, respectively. HBV, hepatitis B virus.
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2017 European Association for the Study of the Liver Terms and Conditions

5. Fig. 3
RG7834 preferentially degrades subgenomic HBV RNA transcripts. (A) Differential expression scores between RG7834 (100 nM) and non-treated, after 6 h (upper panel) and 24 h (lower panel) of treatment, are visualized in grey. A cubic smoothing line with 10 degrees of freedom is fitted for each time point and is shown in red. P, HBV polymerase; S, HBsAg; X, HBx; C, HBV core. (B) Northern blot analysis of 2 μg of RNA extracted from PHH treated five days after HBV infection with RG7834, HBx siRNA (siHBx) or control siRNA (siControl) for the number of hours indicated above the blot. HBV RNA was specifically detected with a genome length HBV (-) strand RNA probe. The migrations of the pgRNA, preS1 and preS2 RNA species are indicated on the right. rRNA was stained on the blotting membrane with methylene blue, the 18S rRNA band served as a loading control. HBV, hepatitis B virus; HBsAg, hepatitis B surface antigen; pgRNA, pregenomic RNA; PHH, primary human hepatocytes; rRNA, ribosomal RNA.
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2017 European Association for the Study of the Liver Terms and Conditions

6. Fig. 4
Effect of RG7834 and ETV on HBV viral markers in serum and liver from HBV genotype C infected uPA/SCID mice. Kinetics of HBV DNA, HBsAg, HBeAg and blood human albumin were determined in serum from vehicle or drug treated HBV-infected uPA/SCID mice during the treatment and follow-up period up to 35 days. Shown are mean values ± standard errors from four to six mice. At day 21, two animals from each group were sacrificed and the livers were harvested for hepatic HBV DNA and cccDNA determination. The remaining animals were monitored for the follow-up period. Log10 reduction p values were calculated using a linear mixed effect model using R software and p values were globally adjusted for all performed comparisons (Holm’s method). cccDNA, covalently closed circular DNA; ETV, entecavir; HBV, hepatitis B virus; HBeAg, hepatitis B e antigen; HBsAg, hepatitis B surface antigen; uPA/SCID, urokinase-type plasminogen activator/severe combined immunodeficiency; n.a., not applicable; QD, once daily; BID, twice daily. aMean log10 reduction ± standard error from baseline; ***p <0.001; *p <0.05; n.s., not significant. bMean log10 reduction ± standard error from vehicle; **p <0.01; n.s., not significant.
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2017 European Association for the Study of the Liver Terms and Conditions

7. Fig. 5
Effect of RG7834, ETV and PegIFNα alone and in combination on HBV viral markers in serum and liver from HBV-infected uPA/SCID mice. Kinetics of HBV DNA, HBsAg, HBeAg and blood human albumin were determined in serum from vehicle or drug treated HBV-infected uPA/SCID mice during the treatment (five mice per group) and follow-up period (three mice per group) up to 49 days. Mean values ± standard errors are shown. ETV, entecavir; HBV, hepatitis B virus; HBeAg, hepatitis B e antigen; HBsAg, hepatitis B surface antigen; PegIFNα, pegylated interferon α-2a; uPA/SCID, urokinase-type plasminogen activator/severe combined immunodeficiency.
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2017 European Association for the Study of the Liver Terms and Conditions

8. Fig. 6
Effect of RG7834, ETV and PegIFNα alone or in combination on HBV viral markers in livers from HBV-infected uPA/SCID mice. HBsAg (red) and cell nuclei (blue) (upper panel), and HBcAg (green), cell nuclei (blue) and human hepatocytes (red) (lower panel) were detected by immunohistochemistry in the mouse liver samples at the end of treatment (day 35). ETV, entecavir; HBV, hepatitis B virus; HBcAg, hepatitis B core antigen; HBsAg, hepatitis B surface antigen; PegIFNα, pegylated interferon α-2a; uPA/SCID, urokinase-type plasminogen activator/severe combined immunodeficiency.
Journal of Hepatology  , DOI: ( /j.jhep )
Copyright © 2017 European Association for the Study of the Liver Terms and Conditions