Evaluation of the drug disposition of RO7049389 with in vitro data and human mass balance supported by physiologically based pharmacokinetic modeling.

RO7049389 (linvencorvir) is a developmental oral treatment for chronic hepatitis B virus infection. The aim of this work was to conduct mass balance (MB) and absolute bioavailability (BA) analyses in healthy volunteers, alongside in vitro evaluations of the metabolism of RO7049389 and a major circulating active metabolite M5 in human hepatocytes, and physiologically based pharmacokinetic (PBPK) modeling to refine the underlying drug disposition paradigm. Participants in the clinical study (MB: Caucasian, male, n=6; BA: Caucasian and Asian, male and female, n=16, 8 in each ethnic groups) received oral [ 14 C] or unlabeled RO7049389 (600/1000 mg) followed by 100 μg intravenous [ 13 C]RO7049389. Metabolic pathways with fractions metabolized - obtained from the in vitro incubation results of 10 μM [ 14 C]RO7049389 and 1 μM M5 with (long-term co-cultured) human hepatocytes in the absence and presence of the cytochrome P450 3A4 (CYP3A4) inhibitor itraconazole - were used to complement the PBPK models, alongside the clinical MB and BA data. The model performance in predicting the pharmacokinetic profiles of RO7049389 and M5 aligned with clinical observations in Caucasians and was also successfully applied to Asians. Accordingly, the drug disposition pathways for RO7049389 were postulated with newly characterized estimates of the fractions: biliary excretion by P-glycoprotein (~41%), direct glucuronidation via uridine 5'-diphosphoglucuronosyltransferase 1A3 (~11%), hexose conjugation (~6%), oxidation by CYP3A4 (~28%) and other oxidation reactions (~9%). These results support the ongoing clinical development program for RO7049389 and highlight the broader value of PBPK and MB analyses in drug development.