Study Of Utility And Mechanism Of Sorafenib Analogues In Mouse Liver Fibrosis

Lacking of effective treatment,hepatic fibrosis has a high rate of morbidity and mortality worldwide.It is increasingly recognized that epithelial-mesenchymal transition?EMT?is a key step for fibrogenesis,in which transforming growth factor-??TGF-??is a pivotal inducer.As a star molecule in cancer therapy,sorafenib is the first oral multi-kinase inhibitor approved by the Food and Drug Administration?FDA?for the clinical treatment of a variety of tumor types.Aside from the established clinical benefits of sorafenib,this drug exerts its anti-fibrotic property in several animal models of organ fibrosis.However,the clinical application of sorafenib is limited due to high cost and multiple adverse drug reactions.Herein,we aim to initially design a number of sorafenib analogues with high activity,low toxicity,and subsequently evaluate their effects both in vitro and in vivo.Certainly,this study will shed light on the therapeutic potential of sorafenib analogues in the clinical treatment of hepatic fibrosis.Initially,we designed a number of sorafenib analogues and found the inhibitory effects on TGF-? signaling were preserved when the kinase binding sites were destroyed.In the modification of the structure of sorafenib,we subsequenyly found carbanilide significantly reduced the TGF-? signaling.Eventually,we found out triclocarban with carbanilide from the list of FDA approved drugs and demonstrated that triclocarban blocked TGF-? signaling in a dose-dependent manner.When applied an ideal cell model in vitro,we showed that treatment with triclocarban counteract TGF-?1-induced concomitant EMT and apoptosis in mouse hepatocytes.Triclocarban reversed the expression profiles of EMT markers,as epithelial marker E-cadherin was up-regulated and the expression level of the mesenchymal gene fibronectin was decreased.We further showed that triclocarban negatively regulated the expression of other EMT related transcription factors,including Snail,ZEB1,and ZEB2 as well as TGF-?1.In addition,we observed that triclocarban significantly inhibited the activation of LX-2hepatic stellate cells induced by TGF-?1 and markedly decreased the expression of TGF-?1?Col I?Col? and TIMP1 in LX-2 cells.To explore the therapeutic potential of triclocarban for liver fibrosis in vivo,we established a hepatic fibrosis model induced by CCl₄ in BALB/c mouse.The rate of mouse liver weight to body weight was decresed and the levels of ALT and AST were significantly lower in triclocarban-treated group.As determined by the H&E and Sirius red staining,the deposition of collagen fibers was largely reduced after treatment with triclocarban.Likewise,a profound decrease ina-SMA expression was observed in the triclocarban-treated group compared with the CCl₄-treated group.In summary,we demonstrate that triclocarban inhibits TGF-?1-induced EMT and apoptosis in mouse hepatocytes and ameliorates CCl₄-induced liver fibrosis in mouse,suggesting an attractive pharmacological tool for the treatment of liver fibrosis and other fibrotic disorders.