Some new transition metal chelates ([M(FSH)2] and [M(FSH) (CH3COO− or NO3−)], where M = Pd2+, VO2+, Mn2+, Zn2+, and cations) incorporating ferrocenyl thiophenol imine ligand (FSH) were prepared. Various spectral and physicochemical studies were performed to elucidate the geometric structure of the investigated compounds. The spectral data of FSH imine ligand and its metal chelates were explained concerning the structural changes due to complex formation. From the electronic spectra and the magnetic moments, the information about geometric structures can be concluded. The activation thermodynamic parameters of the thermal degradation for FSH complexes were calculated utilizing the method of Coats–Redfern. Correlation of all physicochemical tools employed in the investigation and DFT calculation, FSH imine ligand acts as a bidentate ligand and coordinates to Mn2+ in octahedral geometry, VO2+ in square pyramidal geometry, Pd2+ in square planner geometry and Zn2+ in distorted octahedral geometry. in vitro antimicrobial, anticancer and antioxidant activities of FSH ligand and its complexes were checked. All complexes exhibited superiority on the free ligand in successful treatment, specifically the FSHPd complex. The new complexes were subjected to study their DNA binding via various methods such as spectrophotometry, viscosity and gel electrophoresis. Their binding feature to ctDNA was proposed to be electrostatic, intercalation or replacement mode. Finally, the molecular docking studies were performed to understand the essence of the protein synthesized compounds' binding and binding affinity (PDB:3hb5).