Fig. 5. Influence of diseased state on ADME profile of RO6889678. (A) Albumin production of hepatocytes in the coculture HepatoPac model in healthy and (hepatitis B virus HBV)-infected states. No influence of HBV infection on albumin production of hepatocytes could be observed with time (infected hepatocytes, circles; and mock-infected hepatocytes, squares). In the controls, mock-infected fibroblasts (downward triangles), and HBV-infected fibroblasts (upward triangles), only very low albumin levels (background signal) were observed. (B and C) Levels of HBeAg (B) and HBsAg (C) versus time in the HBV-infected coculture HepatoPac model (circles). In the controls, mock-infected hepatocytes (squares), mock-infected fibroblasts (downward triangles), and HBV-infected fibroblasts (upward triangles), no antigen levels (background signal) were observed. (D) Immunostaining of HBV infection in HepatoPac (day 12 postinfection). Nuclear staining was performed with Hoechst 33342 (blue). Immunostaining of HBsAg protein (green) and HB core (red) with secondary Alexa Fluor antibodies. (E and F) Induction potential of RO6889678 in healthy and diseased states (HBV-infected hepatocytes). The fold changes of mRNA expression levels are shown in diseased versus healthy states and after multiple treatments with RO6889678 compound (at day 12 postinfection). (E) Transporter mRNA expression levels of OATP and ABCG2 (BCRP) were increased 2- to 3-fold in healthy and diseased states after RO6889678 treatment. (F) Metabolic enzyme mRNA levels. Two- to 3-fold elevation of CYP3A4, CYP2B6, CYP2C8, CYP2C9, and CYP1A2 expression was observed relative to untreated cells in healthy and disease states. Changes in expression levels (≤2-fold) were found for CYP2D6, UGTs (UGT1A1, UGT1A3), FMOs (FMO3, FMO4, and FMO5), NAT monooxygenases (NAT1 and NAT2), aldehyde oxidase, and aldo-keto reductase. Thus, no changes in induction potential of RO6889678 could be observed comparing healthy and diseased states. D, diseased; H, healthy; RO, RO6889678.