Hinokinin, a lignan from <i>Piper cubeba</i> fruits induces apoptosis via reactive oxygen species-mediated mitochondrial dysfunction in MCF-7 breast cancer cells

The phytochemical investigation of fruits of <i>P. cubeba</i> led to the isolation of sixteen compounds (<b>1-16</b>), including two <b>8</b> and <b>10</b> for the first time. The structures were established using spectroscopic analysis, including (¹H, ¹³C, 2D NMR data and MS analysis). The crude extract, fractions and isolated compounds were tested for cytotoxicity against four human cancer cell lines, Viz. MDA-MB-231, MCF-7, HCT-116 and Mia-PaCa-2. Chloroform fraction exhibited significant cytotoxicity against MCF-7 cancer cell line, whereas compounds (<b>2</b>), hinokinin, and (<b>6</b>) 9S-O-methyl cubebin showed maximum cytotoxicity of 50-40% inhibition at 10 µM concentration against MCF-7, MDA-MB231, and Mia-PaCa-2. Hinokinin (<b>2</b>) didn't show any toxicity up to 100 μM concentration. The mechanistic assays revealed its role in inducing apoptosis in MCF-7 cells via ROS generation and MMP disruption. These results highlight <i>P. cubeba</i> as a source for anticancer compounds, warranting further research into the therapeutic applications of these bioactive substances in cancer treatment.