Study of interaction between synthetic spiropyrimidines (SP) with bovine serum albumin (BSA) was investigated by spectroscopy. The fluorescence of BSA was quenched by SP by means of static quenching mechanism. Using Stern–Volmer analysis of the fluorescence quenching data number of binding sites (n) and binding constants (K) at different temperatures were computed. The thermodynamic parameters enthalpy change (ΔH) and entropy change (ΔS) were calculated using Van’t Hoff equation (ΔH = -95.16 kJ/mol and ΔS = -251.49 J/mol/ K) and the results clearly indicated binding process is enthalpy-driven but entropically disfavored. The weak force van der Waals interaction and hydrogen bonding is responsible for BSA-SP complexation. Synchronous fluorescence, UV-Vis and circular dichroism (CD) spectra proposed possibility of alteration in conformation of BSA in the presence of SP. Using Forster’s non-radiation energy transfer (FRET) theory distance between the donor and the acceptor were found to be <7 nm.