Ex vivo Effects of Sorafenib and Regorafenib on Murine Hepatocytes

Abstract

Sorafenib and regorafenib are structurally-related small-molecular-weight inhibitors of cellular kinases. Regorafenib has a Boxed Warning stating: "Severe and sometimes fatal hepatotoxicity has been observed in clinical trials", while sorafenib is considered less hepatotoxic. This ex vivo study assessed the effects of sorafenib (2.5 and 50 µM) and regorafenib (5.0 and 50 µM) on liver structure, ultrastructure, cellular respiration (mitochondrial O 2 consumption), ATP, caspase activity, urea synthesis, and glutathione. Liver fragments from Taylor Outbred mice were incubated in Krebs-Henseleit buffer (continuously gassed with 95% O 2 :5% CO 2 ) with and without the drugs for 3 to 4 h. The presence of sorafenib or regorafenib had insignificant effects on liver structure, cellular respiration, ATP, caspase-3 activity, urea synthesis, and glutathione. At 3 h, liver histology with and without 2.5 µM sorafenib or 5.0 µM regorafenib was similar. Liver histology with 50 µM sorafenib was slightly worse than untreated tissue at 3 h, showing single hepatocyte necrosis and cellular disintegration. With 50 µM regorafenib, the histology was closely mirroring untreated tissue at 3 h. Similarly, caspase-3, caspase-9, cytochrome c, BAX and annexin A2 immunostains showed no significant drug effects at 4 h (2.5 µM sorafenib or 5.0 µM regorafenib). Electron microscopy revealed a more prominent loss of rough endoplasmic reticulum (rER) integrity with regorafenib treatment compared with sorafenib treatment. Thus, derangements in the rER were more prominent with regorafenib. Otherwise, the studied hepatic surrogate biomarkers did not distinguish between the two compounds.

Isomorphous Crystals Formed by the Similar Supramolecular Motifs in Sorafenib Hydrochloride and Regorafenib Hydrochloride Salts

“…Sorafenib and Regorafenib have been used as the target therapy for the treatment of a large range of tumor types [6]. They inhibit tumor cell growth, proliferation, and tumor angiogenesis [7,8].…”