Four novel dinuclear Cu(II) complexes (1a-4a) have been prepared from ONO-tridentate chiral Schiff base derivatives of chloralose (1-4). All the new complexes (1a-4a) have been characterized by Fourier transform infrared spectroscopy, UV-visible spectroscopy and elemental analysis. The complex 2a was further characterized by X-Ray crystallography. And also, the magnetic susceptibility measurement of this complex (2a, mu(eff) 2.61 BM/dimer) was measured at room temperature. The activities of ONO-tridentate chiral Schiff base derivatives of chloralose (1-4) and their dinuclear Cu(II) complexes (1a-4a) on viability of human cervical cancer cell line (HeLa) were investigated by MIT assay. The obtained results indicated that the Schiff base ligand 1 and its dinuclear Cu(II) complex (1a) have excellent cytotoxic activity. Also, dinuclear Cu(II) complexes of the galactochloralose and alpha-chloralose derivative Schiff base ligands were screened for their in vitro antimicrobial activity against eight bacterial species as well as yeast species Candida albicans ATCC 10231 using disc diffusion method. Complex 4a exhibited promising in vitro antimicrobial activity with inhibition zone diameter from 10 to 22 mm against the tested Gram-positive bacterial strains and yeast C. albicans.