Integrative Computational and Bioactivity-Guided Strategies for Isolating Cytotoxic Phytochemicals in Ginger ( <i>Zingiber officinale</i> Roscoe)

Objective Breast cancer stands as one of the most prevalent cancers globally, representing around 31% of new cancer diagnoses in women. The emergence of drug resistance undermines the efficacy of chemotherapy, highlighting the urgent need for alternative treatments. Ginger ( Zingiber officinale ) is a dietary plant known for its different biological effects. Our goal was to isolate the cytotoxic compounds from the petroleum ether extract of dried ginger. Methods Through bioactivity-guided isolation and comprehensive phytochemical analyses, the cytotoxic effects of different concentrations (ranging from 3.91 to 500 µg/mL) of ginger petroleum ether extract and its fractions were examined against MCF-7, SW 480, and A549 cancer cell lines, as well as 3T3 normal cells using the MTT assay. Fractions were obtained using column chromatography, and those showing significant toxicity were further fractionated using a bioactivity-guided separation method. The most cytotoxic component was purified using plate chromatography, and its toxicity was evaluated. Additionally, molecular docking was performed on gamma-secretase, PPARγ, MMP-9, tubulin, proteins Notch1, HES1, and cyclin D1. Results Fractions showed toxicity against all the tested cell lines, with a greater effect on MCF-7. A subfraction, FsA7, exhibited an impressive IC 50 value of 1.68 ± 0.36 µg/mL against MCF-7. It also showed selective toxicity towards cancer cells compared to 3T3 cells. Upon analysis using GC/MS, it was identified as 6-shogaol based on the retention index (RI) and library search programs. Molecular docking studies indicated that 6-shogaol interacts with critical cancer-related targets, especially MMP-9, shedding light on its mechanism of action. Conclusion These findings highlight the therapeutic potential of 6-shogaol as a complementary treatment in breast cancer. This research provides foundational insights into the use of ginger-derived compounds in enhancing current cancer treatment strategies. Pre-clinical and clinical evaluation of cytotoxic effects of 6-shogaol in combination with anti-cancer drugs can be considered for future studies.