Fluorofenidone is an Effective Drug for Cholestasis and Fibrosis in a Preclinical Study

Background: Cholestasis is characterized by intrahepatic accumulation of bile acids (BAs), resulting in liver injury, fibrosis, and liver failure. To date, only ursodeoxycholic and obeticholic acids have been approved for the treatment of cholestasis. As fluorofenidone (AKF-PD) was previously reported to play significant anti-fibrotic and anti-inflammatory roles in various diseases, we investigated whether AKF-PD ameliorates cholestasis. Methods: A mouse model of cholestasis was constructed by administering a 0.1% 3,5-diethoxycarbonyl-1,4-dihydroxychollidine (DDC) diet for 14 days. Male C57BL/6J mice were treated with either AKF-PD or pirfenidone (PD) orally in addition to the DDC diet. Serum and liver tissues were subsequently collected and analyzed. Findings: We found that AKF-PD significantly reduced the levels of serum ALT, AST, and TBA, as well as hepatic BA levels. Hepatic histological analyses demonstrated that AKF-PD markedly attenuated hepatic inflammation and fibrosis. Further preliminarily mechanistic analyses revealed that AKF-PD markedly inhibited expression of Cyp7a1, an enzyme key to BA synthesis, by increasing FXR nuclear translocation, and decreased hepatic inflammation by attenuating Erk/Egr-1-mediated cytokine and chemokine expression. Moreover, AKF-PD was found to substantially reduce liver fibrosis via inhibition of TGFβ1 expression in our mouse model. Interpretation: Here, we found that AKF-PD effectively attenuates cholestasis and hepatic fibrosis in a mouse model of DDC-induced cholestasis. As such, AKF-PD warrants further investigation as a candidate drug for treatment of cholestasis. Funding The National Natural Science Foundation of China; the Outstanding Youth Science Foundation of Chongqing; the Project of Chongqing Universities Innovation Research Group.Declaration of Interest: The authors declare no competing interests.Ethical Approval: All animal experiments were approved by the animal care and use committees at the Medical Research Center (Southwest Hospital, Chongqing, China; the approvednumber: AMUWEC20223018).