Spirothiazolidines were versatile synthetic scaffold processing wide spectrum of biological interests involving potential anticancer activity.Report the first synthesis of Bis Spiro-thiazolidene as a novel heterocyclic ring system.One-pot three-component reaction include condensation of p-phenyllene diamine; cyclohexanone and thioglycolic acid produced Spiro-thiazolidine 4 which undergo further condensation with cyclohexanone and thioglycolic acid with eqiumolar ratio to introduce Bis-Spiothiazolidine 5 as a first synthesis. Also;bis spiro-thiazolidine arylidene derivatives 6‒13 were synthesized by reaction of Bis-Spiothiazolidine 5 with different aromatic benzaldehyde.Four compounds 13, 12, 9 and 11 have showed high significantly anticancer activity compared to Doxorubicin® (positive control) against Human liver carcinoma (HepG2) and human normal Retina pigmented epithelium (RPE-1) cell lines.The novel bis-spirothiazolidine deriviatives synthesized for the first time and showed excellent anticancer activities.