Fig 2: Overexpression of ASGPR-1/-2 Increases Antiviral Activity of Anti-HBV LNA-SSO upon GalNAc Conjugation(A and B) Inhibition of HBsAg and HBeAg secretion and intracellular mRNA levels in HBV-infected dHepaRG-WT (A) and dHepaRG-ASGPR-1/-2 (B) cells after treatment with anti-HBV LNA-SSO (SSO-1), GalNAc-conjugated anti-HBV LNA-SSO (SSO-2), and control LNA-SSO (SSO-3). Error bars, SD (n = 3). (C) Table represents the average EC50 values for all measured anti-HBV markers in both cell lines of three independent experiments.

Fig 3: The GalNAc-Conjugated SSO-2 Antiviral Activity Is Target Sequence Dependent(A) Inhibition of HBsAg secretion was measured by CLIA from supernatant of HBV-infected dHepaRG ASGPR1/2 after treatment with GalNAc-conjugated HBV LNA-SSO as SSO-2 and its variants harboring either one or two mismatches, SSO-4 and SSO-5, respectively. Error bars, SD (n = 3). (B) Table represents the average EC50 values for inhibition of HBsAg production from three independent experiments. (C) AAV-HBV mice were treated with saline (control), SSO-2, SSO-4, or SSO-4 at 0.18 µmol/kg on days 0, 3, 6, and 9. Serum HBsAg was determined during the treatment and follow-up period. Error bars, SEM (n = 4). (D) The reduction in circulating HBsAg and HBV DNA at day 16 (one week after treatment was stopped) is summarized in a table. Dose level of 0.18 µmol/kg corresponds to 1.4 mg/kg of SSO-2, SSO-4, and SSO-5.