This study presents the synthesis, structural characterization, and in vitro evaluation of anticancer activity of some newly benzo quinoline derivatives. The synthesis is facile and efficient, involving two steps: quaternization of nitrogen heterocycle followed by a 3+2 dipolar cycloaddition reaction. The synthesized compounds were characterized by FTIR, NMR, and X-ray diffraction on monocrystal in the case of compounds and . An in vitro single-dose anticancer assay of eighteen benzo quinoline compounds, quaternary salts, and cycloadducts, was performed. The results showed that the most active compounds were quaternary salts and with aromatic R substituents. Quaternary salt revealed non-selective activity against all types of cancer cells, while salt exhibited a highly selective activity against leukemia cells. Compound also presented remarkable cytotoxic efficiency against four distinct types of cancer cells-namely, non-small cell lung cancer HOP-92, melanoma LOX IMVI, melanoma SK-MEL-5, and breast cancer MDA-MB-468. Compound was selected for five-dose screening. The study also includes SAR correlations.