Amorfrutin C Induces Apoptosis and Inhibits Proliferation in Colon Cancer Cells through Targeting Mitochondria

A known (<b>1</b>) and a structurally related new natural product (<b>2</b>), both belonging to the amorfrutin benzoic acid class, were isolated from the roots of <i>Glycyrrhiza foetida</i>. Compound <b>1</b> (amorfrutin B) is an efficient agonist of the nuclear peroxisome proliferator activated receptor (PPAR) gamma and of other PPAR subtypes. Compound <b>2</b> (amorfrutin C) showed comparably lower PPAR activation potential. Amorfrutin C exhibited striking antiproliferative effects for human colorectal cancer cells (HT-29 and T84), prostate cancer (PC-3), and breast cancer (MCF7) cells (IC₅₀ values ranging from 8 to 16 μM in these cancer cell lines). Notably, amorfrutin C (<b>2</b>) showed less potent antiproliferative effects in primary colon cells. For HT-29 cells, compound <b>2</b> induced G0/G1 cell cycle arrest and modulated protein expression of key cell cycle modulators. Amorfrutin C further induced apoptotic events in HT-29 cells, including caspase activation, DNA fragmentation, PARP cleavage, phosphatidylserine externalization, and formation of reactive oxygen species. Mechanistic studies revealed that <b>2</b> disrupts the mitochondrial integrity by depolarization of the mitochondrial membrane (IC₅₀ 0.6 μM) and permanent opening of the mitochondrial permeability transition pore, leading to increased mitochondrial oxygen consumption and extracellular acidification. Structure–activity-relationship experiments revealed the carboxylic acid and the hydroxy group residues of <b>2</b> as fundamental structural requirements for inducing these apoptotic effects. Synergy analyses demonstrated stimulation of the death receptor signaling pathway. Taken together, amorfrutin C (<b>2</b>) represents a promising lead for the development of anticancer drugs