Evaluation of the genetic toxicity of sofosbuvir and simeprevir with and without ribavirin in a human-derived liver cell line.

Carina S Librelotto,Rafael R Dihl,Mário R Álvares-Da-Silva,Ana Paula De Souza,Daniel Simon

doi:10.1590/0001-3765202120200632

Abstract

Direct-acting antivirals have revolutionized the treatment of chronic hepatitis C. Sofosbuvir and simeprevir are prescribed worldwide. However, there is a scarcity of information regarding their genotoxicity. Therefore, the present study assessed the cytotoxic and genotoxic effects of sofosbuvir and simeprevir, alone and combined with ribavirin. HepG2 cells were analyzed using the in vitro cytokinesis-block micronucleus cytome assay. Cells were treated for 24 h with sofosbuvir (0.011-1.511 mM), simeprevir (0.156-5.0 µM), and their combinations with ribavirin (0.250-4.0 mM). No significant differences were observed in the nuclear division cytotoxicity index, reflecting the absence of cytotoxic effects associated to sofosbuvir. However, the highest concentration of simeprevir showed a significant difference for the nuclear division cytotoxicity index. Moreover, significant results were observed for nuclear division cytotoxicity index in two combinations of sofosbuvir plus ribavirin and only in the highest combination of simeprevir plus ribavirin. Additionally, our results showed that sofosbuvir did not increase the frequency of chromosomal damage, but simeprevir significantly increased the frequency of micronuclei at the highest concentrations. The combination index demonstrated that both sofosbuvir and simeprevir produced antagonism to the genotoxic effects of ribavirin. In conclusion, our results showed that simeprevir, but not sofosbuvir, has genotoxic effects in HepG2 cells.

Highlights

The combined treatment of SOF + RBV (Figure 2a) was able to reduce nuclear division cytotoxicity index (NDCI) values compared to the negative control, and this reduction was statistically significant for two combined treatments (SOF + RBV: 0.755 + 2 and 1.511 + 4 mM)
MNi, nucleoplasmic bridges (NPBs), and nuclear buds (NBUDs) frequencies were investigated in HepG2 cells using the CBMNCyt assay (Tables I and II)
When comparing the induction frequencies of both treatments at each concentration, it can be seen that the combined treatment (SIM + RBV) led to significant increases in frequency of MNis compared with the treatment with SIM alone

Summary

INTRODUCTION

Chronic hepatitis C affects approximately 71 million people worldwide, or about 1% of the world’s population (WHO 2017). The approval of SOF by the Food and Drug Administration (FDA) represented a milestone in the development of new therapeutic options, and created new perspectives in the development of potent treatment regimens for chronic hepatitis C. These new DAAs and their combinations revolutionized HCV treatment, with shorter treatment regimens, lower toxicity, and more effective response (SVR > 90%) (Geddawy et al 2017). The present study assessed the cytotoxic and genotoxic potentials of two DAAs (SOF and SIM), with and without RBV, using the in vitro cytokinesis-block micronucleus cytome (CBMN-Cyt) assay in human hepatoma cells (HepG2)

MATERIALS AND METHODS

RESULTS

DISCUSSION