For the development of new anticancer agents, dibenzo[(γ-aryl)pyridino]aza-17-crown-5 ethers containing 2,4,6-triarylpyridine were synthesized successfully by one-step domino-condensation of 1,8-bis(2-acetylphenoxy)-3,6-dioxaoctane, arylaldehydes and ammonium acetate according to the conditions of Hantzsch reactions. The synthesized γ-arylpyridine derivatives show high cytotoxic activity against human cancer cell lines: Hep-G2, RD, FL, Lu1. Compounds (3b,c) showed significant cytotoxicity against all four human cell lines whereas the similar synthesized compound (3d) possessed cytotoxicity against HepG2, Lu1 and RD cell lines. Meantime, both compounds (3f,g) containing γ-heteroaryl only exhibited cytotoxicity against RD and FL cell lines. Azacrown ethers (3b-d) exhibited low activity on the Vero cell line, meaning that they can be evaluated for their potential as promising anticancer agents. X-Ray structure study was performed to determine the structure of the representative compound 3a. © ISUCT Publishing.