Preparation and anti-tumor activity of natural product genistein imidazole derivatives

Genistein, a natural isoflavone, exhibits potential anti-tumor properties but suffers from weak cytotoxicity that limits its clinical application. To improve its antitumor activity, five genistein imidazole/triazole derivatives (G1–G5) were designed and synthesized via molecular hybridization method in this study, four of them are new compounds that have not been reported in the literature. The in vitro antiproliferative effects of all target compounds against seven human cancer cell lines (HepG2, HeLa, A549, SH-SY5Y, DU145, PC-3 and MCF-7) were evaluated by MTT assay, with 5-fluorouracil (5-Fu) and the parent genistein as positive controls. The results demonstrated that all synthesized derivatives possessed significantly enhanced tumor-suppressive activity compared with genistein, and most compounds have better inhibitory activity against tested cancer cells than 5-Fu. Among them, compound G5 displayed the strongest broad-spectrum antitumor capacity with the lowest IC₅₀ values. Further experimental study on Mechanism of action confirmed that G5 induced HeLa cell apoptosis in a dose-dependent manner and triggered typical apoptotic morphological changes. Molecular docking analysis revealed that G5 could tightly bind to CDK2 and FAK with binding free energies (−8.14 and − 9.37 kcal/mol, respectively), forming stable complexes to inhibit enzyme activities. Structure-activity relationship analysis indicated that the introduction of triazole moiety contributed more to the antitumor potency than imidazole groups. This work proves that structural modification with imidazole/triazole heterocyclic groups is an effective strategy to enhance the antitumor activity of genistein. Compound G5 may serve as a promising dual-target lead candidate targeting CDK2 and FAK for the development of novel anti-cancer agents.