N‐Propargylic β‐enaminones in breast cancer cells: Cytotoxicity, apoptosis, and cell cycle analyses

Abstract

Breast cancer is one of the most common cancers worldwide and the discovery of new cytotoxic agents is needed. Enaminones are regarded to be a significant structural motif that is found in a variety of pharmacologically active compounds however the number of studies investigating the anticancer activities of N-propargylic β-enaminones (NPEs) is limited. Herein we investigated the potential cytotoxic and apoptotic effects of 23 different NPEs (1-23) on human breast cancer cells. Cytotoxicity was evaluated via MTT assay. Apoptotic cell death and cell cycle distributions were investigated by flow cytometry. CM-H2DCFDA dye was used to evaluate cellular ROS levels. Expression levels of Bcl-2, Bax, p21, and Cyclin D1 were measured by quantitative real-time PCR. ADME properties were calculated using the ADMET 2.0 tool. NPEs 4, 9, 16, and 21 showed selective cytotoxic activity against breast cancer cells with SI values >2. NPEs induced apoptosis and caused significant changes in Bcl-2 and Bax mRNA levels. The cell cycle was arrested at the G0/G1 phase and levels of p21 and Cyclin D1 were upregulated in both breast cancer cells.ROS levels were significantly increased by NPEs, suggesting that the cytotoxic and apoptotic effects of NPEs were mediated by ROS. ADME analysis revealed that NPEs showed favorable distributions in both breast cancer cell lines, meaning good lipophilicity values, low unfractionated values, and high bioavailability. Therefore, these potential anticancer compounds should be further validated by in vivo studies for their appropriate function in human health with a safety profile, and a comprehensive drug interaction study should be performed.

Evaluation of Novel Spiro-pyrrolopyridazine Derivatives as Anticancer Compounds: In Vitro Selective Cytotoxicity, Induction of Apoptosis, EGFR Inhibitory Activity, and Molecular Docking Analysis

“…By assessing fluorescence patterns, viable cells (annexin V- and PI-) and apoptotic cells (annexin V+ and PI+) can be distinguished, providing insights into their respective percentages. We used FITC Annexin V Apoptosis Detection Kit I (BD Pharmingen) . Briefly, cells were seeded at a density of 10 6 cells per well in a six-well plate and treated with the most effective concentrations of SPPs for 72 h. Subsequently, cells were washed with cold PBS, resuspended in 1 mL of 1× Binding Buffer, and stained with 5 μL of Annexin V FITC and 5 μL of PI.…”

“…We used FITC Annexin V Apoptosis Detection Kit I (BD Pharmingen). 15 Briefly, cells were seeded at a density of 10 6 cells per well in a six-well plate and treated with the most effective concentrations of SPPs for 72 h. Subsequently, cells were washed with cold PBS, resuspended in 1 mL of 1× Binding Buffer, and stained with 5 μL of Annexin V FITC and 5 μL of PI. After vortexing, the solution was incubated at room temperature (25 °C) for 15 min in the dark.…”