Evaluation of molecular interaction studies of khellin on bovine serum albumin through various biophysical approaches

The understanding of how drugs interact with carrier proteins is crucial in the field of pharmacology and the life sciences, especially in the field of drug invention. In the present work described the molecular interaction of pharmaceutically important phyto-molecule khellin on bovine serum albumin. Khellin is recognized for its ability to widen blood vessels, making it useful for heart health. It's a major component of the plant Ammi visnaga and Dioscorea species helps to protect the heart. Bovine serum albumin (BSA) is a model protein of Human serum albumin hence, BSA has been used for drug-binding properties studies. Various biophysical techniques to examine the interactions between khellin and BSA. The biophysical techniques such as fluorescence quenching by fluorescence spectroscopy studies, micro-environmental changes by synchronous fluorescence, protein structural changes by circular dichroism spectroscopy, molecular docking, ADMET properties studies, SWISS Target Prediction for target analysis and pharmacokinetic analysis by insilico . The Khellin-BSA interaction was examined using fluorescence analysis, which showed that the binding constant was 1.29 ± 0.2 × 10¹² M⁻¹ and binding free energy was −7.99 kcal/mol by in vitro . The binding energy was compared with computation molecular docking studies of ligand and protein interaction showed the binding energy of −5.1 kcal/mol. It is nearer to the in vitro binding energy values of khellin on BSA. The micro-environmental changes of the ligand-protein complex were observed with peak shifts at Δλ15 for tyrosine, Δλ60 for tryptophane, and Δλ90 for phenylalanine. Also, the secondary structural changes of BSA after titrating the khellin were observed and found that there were secondary structural changes in the free BSA after adding the khellin. With possible targets found through SWISS Target Prediction, khellin is a promising druggable candidate, according to ADMET analysis, which revealed zero violations. Finally, we concluded that the Phyto-active constituent khellin possesses good binding affinity on BSA. Further, it can be taken for drug discovery experiments on clinical trials.