Two imine compounds HS1 and HS2 derived from Disperse black 9 were prepared and screened for their DNA binding properties using spectrophotometric and in-silico computational studies. To prepare the compounds HS1 and HS2, the 2,4- and 2,5-dihydroxy benzaldehyde compounds containing the propargyl group have been obtained in accordance with the known methods. The compounds have been characterized by the 1H(13C)NMR, mass spectra, UV-vis and FTIR. The electrochemical properties of the compounds have been investigated in the 100-1000 mVs−1 range. The compounds indicate irreversible anodic redox processes at 0.2 V. Single crystals of the compounds have obtained from the methanol solution and their molecular structures have solved by the X-ray method. Thermal behaviours of the compounds have investigated by the thermal analyses methods. The compound HS2 has higher thermal stability than the compound HS1. Both compounds were screened their DNA and BSA binding properties by using spectrophotometric methods. The compounds bind to the minor groove site of DNA with comparable binding constants. Finally, binding interactions and modes of the compounds with DNA and BSA were investigated by molecular docking studies.