Chromanone derivatives demonstrate remarkable cytotoxity against a varieties of cancer cell lines. Novel 9- [hydroxy (substitutedphenyl) methyl] -2, 2-dimethyl-2, 3, 8, 9- tetrahydro- 4H, 10H-pyrano [2, 3-f] chromene-4, 10-diones as Glyasperin analogues were synthesized in four steps from known 4-chromone 1. The key step was the preparation of chromane dione 5a by regioselective intramolecular cyclization reaction in 85% yield. Condensation of 5a with substituted aromatic aldehydes afforded corresponding alpha hydroxybenzyl analogues 6a-6e. The cytotoxic study of the synthesized compounds against breast cancer human cell line (T47D) showed moderate cytotoxic activities (IC₅₀=16-40mM) compared to the positive control drug vincristin (IC₅₀=2.5 mM).