Muhammad Taupik, Nanang Fakhruddin, Mohamad A. Mustapa, Endah N. Djuwarno, Qamar U. Ahmed
Natural products continue to provide valuable leads for anticancer drug discovery, yet limited studies have addressed the bioactive constituents of Spigelia anthelmia Linn. This research aimed to isolate and characterize cytotoxic metabolites of S. anthelmia using a bioassay-guided isolation approach. Sequential extraction, vacuum liquid chromatography (VLC), and preparative thinlayer chromatography (PTLC) yielded a semi-purified fraction designated as Isolate A. Cytotoxicity was evaluated against T47D breast cancer and WiDr colon cancer cells using the MTT assay. Isolate A significantly reduced cell viability in a dose-dependent manner (ANOVA, p < 0.05), with IC₅₀ values of 166.92 ± 5.10 µg/mL (T47D) and 245.24 ± 6.25 µg/mL (WiDr), whereas doxorubicin exhibited IC₅₀ values of 40.05 ± 2.30 µg/mL and 3.57 ± 0.45 µg/mL, respectively. Flow cytometry with Annexin V–FITC/PI staining revealed a visible shift of cell populations toward early and late apoptotic quadrants, indicating apoptosis as the predominant mechanism of cell death. Spectroscopic characterization by FTIR, LC–MS, and NMR suggested that Isolate A is a phenolic/alkaloid-type compound (m/z 218 [M+H]⁺) containing hydroxyl, carbonyl, and heteroaromatic groups. Collectively, these findings demonstrate that S. anthelmia contains apoptosis-inducing metabolites with measurable cytotoxicity, supporting its potential as a natural source of anticancer leads and warranting further purification and mechanistic investigation. To our knowledge, this is the first study to combine bioassay-guided isolation, apoptosis profiling, and spectroscopic characterisation of S. anthelmia metabolites in T47D and WiDr cancer cells. © 2025 Taupik et al.
Department of Pharmacy, Faculty of Sports and Health, Gorontalo State University, Gorontalo, 96211, Indonesia; Department of Pharmaceutical Biology, Faculty of Pharmacy, Gadjah Mada University, Yogyakarta, Indonesia; Department of Pharmaceutical Chemistry, Faculty of Pharmacy, International Islamic University Malaysia, Kuantan, Malaysia
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