MF1, a novel robustic acid derivative, exerts antitumor effects in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) extensive genomic heterogeneity and immunosuppressive tumor immune microenvironment (TIME) are major factors contributing to treatment resistance. This study aims to elucidate the molecular mechanisms by which Robustic Acid 4-O-amino substituted synthetic derivatives (MF1), a novel robustic acid derivative, derived from structural modifications of natural products from Dalbergia benthamii Prain, targets carcinogenic signaling and synergistically regulates the crosstalk between autophagy and ferroptosis to induce programmed cell death in HCC. In vitro functional assays confirmed that MF1 exhibits significant antitumor activity. Mechanistically, MF1 significantly upregulates cellular oxidative stress levels, inducing characteristic changes in mitochondrial ultrastructure and apoptosis; simultaneously, by promoting intracellular Fe²⁺ accumulation and downregulating the expression of key ferroptosis regulators GPX4 and SLC7A11, it potently activates the ferroptosis process. Concurrently, MF1 continuously activates the cytotoxic autophagy pathway by regulating the LC3 I/II conversion ratio and upregulating the expression of core autophagy proteins ATG5 and Beclin-1. Network pharmacology identified 7 core targets of MF1 that are common to both ferroptosis and autophagy regulation. qPCR validation showed that MF1 significantly downregulated the mRNA expression of these 7 core oncogenes in HepG2 cells. Molecular docking confirmed strong binding affinities between MF1 and five core targets, 100 ns all-atom molecular dynamics simulations further validated the conformational stability of all five top-ranked MF1-protein complexes, with the higher binding stability for AKT1, SRC and MTOR. In summary, this study clarifies MF1 induces apoptosis and ferroptosis in HepG2 cells, suggesting its potential as a new therapeutic option for HCC.