In silico and in vitro investigation of anticancer effect of novel anthraquinone compound on MDA-MB-231 cells

Background The serious adverse effects of chemotherapeutics forced to produce new bioactive substances such as anthraquinone derivatives. Objective This study focuses on evaluating the antiproliferative effects of a newly developed anthraquinone compound, referred to as Compound 3, against MDA-MB-231 breast cancer cells. Methods To assess the pharmacokinetic properties of Compound 3, an in silico ADME (Absorption, Distribution, Metabolism, and Excretion) evaluation was carried out. To investigate its binding behavior with cancer-relevant proteins, molecular docking simulations and interaction analyses were performed. For assessing its antitumor activity, a series of biological assays were applied. Cell viability was determined using the MTT assay, while intracellular ROS levels were quantified with the DCFH-DA probe. Apoptotic cell death was evaluated through Annexin V/PI staining, and cell motility was studied via wound healing assays. In addition, alterations in cytoskeletal architecture were analyzed using immunofluorescence microscopy. Results Compound 3 exhibited stronger binding affinities (indicated by lower docking energy values) compared to reference compounds for several targets, including PCNA, CK2, LC3, and MMP2. The in vitro analysis revealed a significant selective inhibitory effect on cellular proliferation along with a marked increase in intracellular ROS levels. Apoptosis was induced by certain doses of Compound 3. Additionally, treatment resulted in decreased migratory behavior and visible disruption of actin cytoskeleton organization. Conclusion The overall findings emphasize the diverse biological effects of Compound 3 on breast cancer cells, revealing promising therapeutic mechanisms for further development as a compelling candidate of an anticancer agent.