Thymoquinone, a potential therapeutic agent of Nigella sativa, binds to site I of human serum albumin

Abstract 

Thymoquinone (TQ) is the main constituent of Nigella sativa essential oil which shows promising in vitro and in vivo antineoplastic growth inhibition against various tumor cell lines. Because of the increasing interest to test it in pre-clinical and clinical researches for assessing its health benefits, we here evaluate the interactions between TQ and human serum albumin (HSA), a possible carrier of this drug in vivo. Binding to HSA was studied using different spectroscopic techniques. Fourier transform infrared (FT-IR) and circular dichroism (CD) spectroscopies suggest that the association between TQ and HSA does not affect the secondary structure of HSA. Using fluorescence spectroscopy, one mole of TQ was found to bind one mole of HSA with a binding constant of 2.39 ± 0.2 10⁴M⁻¹. At 25°C (pH 7.4), van’t Hoff’s enthalpy and entropy that accompany the binding were found to be −10.24kJ/mol⁻¹ and 45J/mol⁻¹K⁻¹ respectively. The thermodynamic analysis of the TQ-HSA complex formation shows that the binding process is enthalpy driven and spontaneous, and that hydrophobic interactions are the predominant intermolecular forces stabilizing the complex. Furthermore, displacement experiments using warfarin and ibuprofen indicate that TQ could bind to site I of HSA, which is also in agreement with the results of the molecular modeling study.

Keywords: Thymoquinone, Nigella sativa, Human serum albumin, Drug-protein complex, FT-IR, Fluorescence quenching, Circular dichroism, Molecular docking

Abbreviations: TQ, thymoquinone, DHTQ, dihydrothymoquinone, DHTQ-GS, glutathionyl-dihydrothymoquinone, HSA, Human serum albumin, CD, Circular Dichroism, FT-IR, Fourier transform infrared, GSH, glutathione, NADH, nicotinamide adenine dinucleotide reduced form, NADPH, nicotinamide adenine dinucleotide phosphate reduced form.