Design, synthesis and biological evaluation of novel β-caryophyllene derivatives as potential anti-cancer agents through the ROS-mediated apoptosis pathway.

As a top-three cancer in global incidence and mortality, colorectal cancer (CRC) urgently demands novel treatments. β-Caryophyllene (β-CP) and its derivatives, a class of sesquiterpenoids with broad anticancer potential, were structurally optimized in this study to enhance efficacy against CRC. Among the synthesized derivatives, AC-7 exhibited potent cytotoxicity and selectivity in HT-29 cells (IC(50) = 3.09 μM, SI = 6.1), comparable to 5-fluorouracil (5-FU, IC(50) = 3.63 μM, SI = 0.4). Network pharmacology and gene enrichment analyses indicated that apoptosis, autophagy, ROS, and NF-κB were key downstream pathways of AC-7, which were later validated experimentally. AC-7 arrested the cell cycle in the G0/G1 phase, promoted autophagy and apoptosis. ROS were identified as having a central role in regulating these related pathways. In vivo studies revealed the significant antitumor and DNA damage activity of AC-7 in a nude mouse model. These findings suggest that AC-7 is a promising candidate for anti-CRC therapy, acting through the ROS-mediated apoptosis pathway.