Abstract 4748: Molecular targets of quinolinone derivatives with anticancer activity

Currently, we are testing chemical libraries of quinolinone derivatives, compounds with high selective cytotoxic activity. These compounds, derived from quinolone antibiotics, are well known for their antibacterial, antiprotozoal, cytotoxic and immunosuppressive effects. 2-phenyl-3-hydroxy-4(1H)-quinolinones were tested by MTT assay for cytotoxic activity and some of them proved IC50 values in sub-micromolar concentrations. Cell cycle and DNA/RNA synthesis were further analysed by flow cytometry. The accumulation of cells was observed predominantly in G1 phase together with significant DNA and RNA synthesis inhibition. The most active derivative was tested in in vivo cancer mice model with using hollow fibres. Hollow fibres were filled by leukemic CEM cell line and were implanted subcutaneously and intraperitoneally to mice. Afterwards, mice were treated by tested compound. Hollow fibres were after 14 days taken from mice and surviving cells were tested by cytotoxic MTT assay. activity of tested quinolinone was comparable to doxorubicin. For protein targets identification, we have synthesized biotinylated molecules of original derivatives. Biotinylated derivatives were immobilized on the surface of streptavidin coated magnetic beads and whole complex (magnetic bead-streptavidin-biotin-quinolinone) was incubated with total cellular lysate of the CEM cell line. Thereafter non-bonded proteins were washed-out and specifically bonded proteins were separated on SDS-PAGE gel. Following silver-staining of the gel we identified sample lines with specifically bonded proteins to biotinylated compound and negative control. Specific proteins were cut off from the gel and trypsinized to get peptides for protein identification by MS analysis (LC-MS/MS and LC-MALDI). Identified targets are proteins associated with translational regulation, glucose metabolism and cytoskeleton. Potential targets were validated by western blot and by other assays (reporter cell models, kinase and enzymatic assays, evaluation in vitro transcription and translation). The work was supported by grants: LC07017, CZ.1.07/2.3.00/20.0009 coming from European Social Fund. Infrastructural part of this project (Institute of Molecular and Translational Medicine) was supported from the Operational Programme Research and Development for Innovations (project CZ.1.05/2.1.00/01.0030). Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4748. doi:1538-7445.AM2012-4748