A novel benzofuran derivative of β-elemene (ZT-22) inhibits hepatocellular carcinoma cell growth via directly targeting HSPA6.

Hepatocellular carcinoma (HCC) is the fifth most common cancer globally and is associated with a poor prognosis. Current therapies for HCC have limited efficacy and require improvement. In our study, the benzofuran derivative of β-elemene (ZT-22) demonstrated enhanced anti-HCC efficacy compared to β-elemene, both in vitro and in vivo. Using network pharmacology, RNA sequencing, and western blot analysis, the crucial role of the p38 MAPK signaling pathway in the anti-HCC activity of ZT-22 cells was highlighted. Using drug affinity-responsive target stability (DARTS) combined with mass spectrometry (MS), HSPA6 was identified as the target for ZT-22. Techniques such as the cellular thermal shift assay (CETSA), surface plasmon resonance (SPR) analysis, microscale thermophoresis (MST), molecular docking and molecular dynamics (MD) simulations were used for further validation, confirming that ZT-22 directly binds to HSPA6. Knocking down HSPA6 diminished p38 MAPK signaling and reversed the anti-HCC effects of ZT-22. These findings suggest that ZT-22 exerts its anti-HCC activity by targeting HSPA6, which in turn activated the p38-MAPK signaling pathway. Our results support the development of ZT-22 as a potential therapeutic agent for HCC and highlight HSPA6 as a promising therapeutic target for HCC treatment.