Design, synthesis, and experimental evaluation of rupestonic acid derivatives as novel anti-tumor agents guided by network pharmacology and molecular docking

This integrated study elucidates the antitumor potential of Artemisia rupestris L.’s primary bioactive component, rupestonic acid (RA), and advances a superior derivative through a systematic, multi-stage strategy. The investigation commenced with network pharmacology, which predicted RA’s polypharmacology mechanism by associating it with 55 potential cancer-related targets—including pivotal nodes like the androgen receptor (AR) and protein kinase C eta (PKCη)—and enriched pathways such as PI3K/AKT and MAPK signaling. In vitro screening across 30 human cancer cell lines validated this prediction, identifying HCT116 and HepG2 as highly sensitive. To enhance efficacy, a structure-based design yielded 27 novel heterocyclic derivatives. Among these, compound 15 emerged as the optimal candidate, demonstrating potent cytotoxicity with IC₅₀ values of 6.203 µM (HCT116) and 9.955 µM (HepG2), significantly surpassing cisplatin. Molecular docking revealed the structural basis for this activity, showing compound 15’s strong binding to key targets like 17β-HSD1 and p38 MAPK via hydrogen bonds and π-π stacking. The stability of these complexes was confirmed through molecular dynamics simulations, which demonstrated convergent RMSD, low binding-site flexibility (RMSF), and sustained favorable interaction energies. Complementing this, a comprehensive in silico ADMET profile established the promising drug-like character of compound 15, predicting high gastrointestinal absorption, no blood–brain barrier permeation, compliance with major drug-likeness rules, and a low toxicity risk. In conclusion, this work from predictive modeling to experimental validation and pharmacokinetic assessment not only deciphers RA’s multi-target mechanism but also identifies compound 15 as a stable, absorbable, and potent lead compound, providing a validated foundation for the development of novel natural product-derived anticancer agents.