The manuscript encompasses the synthesis of four water soluble copper(II) complexes, their structural characterization and a study of biological activities such as binding with CT-DNA, BSA/HSA, antimicrobial and anticancer. Results implied that the complexes showed better activity than the ligands. The antiproliferative activities of both the ligands and complexes on MCF-7 (human breast cancer) and A549 (human lung carcinoma) cell lines were seen to be greater than the standard drug cisplatin. The outcomes suggested complex 3 to be a potential antitumor agent. Display omitted 7-Hydroxy-4-oxo-4H-chromene Schiff bases and their corresponding water soluble copper(II) complexes were synthesized and characterized by various spectral techniques. The structural confirmation of the complexes 2 and 3 has been provided by X-ray crystallographic analysis. The binding affinities of the ligands and complexes with Calf Thymus DNA (CT-DNA) and proteins (Bovine Serum Albumin (BSA) and Human Serum Albumin (HSA)) were studied by using absorption and emission titration methods. An intercalative mode of binding with CT-DNA and a static quenching mechanism with albumin (BSA and HSA) was confirmed from the obtained results. The efficiency of the compounds in inhibiting the microbial growth was tested against various pathogenic bacteria Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pneumoniae and Acinetobacter baumannii by using Gentamicin as standard and fungi Candida albicans, Aspergillus niger, Trichophyton rubrum, Candida tropicalis and Aspergillus fumigatus by using ketoconazole as standard and a significant activity has been found for the complexes. The anticancer potential of the ligands and complexes were tested against A549 and MCF-7 cells by MTT assay using cisplatin as a standard. The complexes 1–4 showed better cytotoxic profile than cisplatin, when incubated for a period of 48 h and 72 h. The results of all the biological analyses indicated the higher activity of the complexes over the ligands. Out of the complexes, complex 3 with the more electron donating ethyl group exhibited the higher activity.