Characterization of the New Pentafluorosulfanyl-Substituted Chalcone 246TMP-3SF5 as Potential New Treatment Option Against Hepatocellular Carcinoma

Background/Objectives: Advanced-stage hepatocellular carcinoma is characterized by a very poor prognosis; thus, highly effective medication is still needed. Often overexpressed heat shock protein 90 is a promising target due to its pivotal role in carcinogenesis. Methods: Antiproliferative effects of novel synthesized inhibitor 246TMP-3SF5 on HepG2 and HuH-7 were analyzed through crystal violet staining. Apoptosis was assessed by measurement of subG1 peak, caspase-3 activity and cleavage of PARP. Ferroptosis was evaluated through reactive oxygen species, glutathione and malondialdehyde levels. Scratch assays were used to assess cancer cell migration and in-ovo models to analyze effects of 246TMP-3SF5 on angiogenesis and microtumors. Molecular docking and molecular dynamics simulation into heat shock protein 90 were carried out using Autodock Vina and Gromacs respectively. Results: Profound dose- and time-dependent antiproliferative effects of 246TMP-3SF5 against HCC cell lines were observed, revealing low micromolecular IC50 values. A significant increase in subG1 peak, key effector caspase-3 activity as well as cleavage of PARP strongly suggested apoptosis playing a crucial role in the antiproliferative effects. Additionally, HuH-7 cells revealed an elevation of reactive oxygen species and both cell lines showed significant glutathione depletion concomitant with malondialdehyde concentration increased upon treatment. The observed effect could be partially reversed applying ferrostatin-1, suggesting ferroptosis as an additionally relevant mode of action. Changes in the cell cycle as well as impaired tumor cell migration were observed. Upon treatment angiogenesis was impaired and mass of microtumors was significantly reduced. Molecular docking revealed interaction with catalytic site of heat shock protein 90. Conclusions: 246TMP-3SF5 is a promising novel inhibitor meriting further research as potential treatment against hepatocellular carcinoma.