Behenic Acid as a multi-target inhibiting antibacterial phytochemical against Vibrio parahaemolyticus and Aeromonas hydrophila for effective management of aquaculture infections: an in-silico, in-vitro & in-vivo experimentation.

Multi-Target Inhibitors are the upcoming frontrunners of the antibiotic world as they provide significant advantage over drug resistance development. Antibacterial drug discovery research, requires more robust and innovative approaches such as multi-target inhibiting drugs, which over comes the innate hurdles in the field of antibiotics. In this current study, a curated set of 5,112 phytochemical molecules were virtually screened for its multi-target inhibition potential against 7 antibacterial protein drug-targets. Behenic Acid was identified to be the most significant phytochemical molecule with potential to inhibit Catalase Peroxidase (KatG), Adenylosuccinate Synthetase (ADSS) and Pyridoxine 5'-Phosphate Synthase (PdxJ), based on SeeSAR and AutoDock Vina results. Further, the inhibition potential of Behenic Acid was validated using 500 ns Molecular Dynamics (MD) Simulation based on Desmond analysis. Behenic Acid was further investigated in-vitro using agar-well-diffusion and Minimal Inhibitory Concentration (MIC) assay, where it demonstrated 20 ± 1mm zone-of-inhibition and 50 µg/ml MIC value against both Vibrio parahaemolyticus and Aeromonas hydrophila. Zebrafish based investigations was carried to confirm the in-vivo antibacterial efficacy of Behenic Acid. It was observed that, there is a progressive dose-dependent recovery from the bacterial infection, with highest recovery and survival observed in fishes fed with 100 µg/day of Behenic Acid. Results of the in-vitro and in-vivo assays strongly support the in-silico prediction of the antibacterial activity of Behenic Acid. Based on the results presented in this study, it is concluded that, Behenic Acid is a strong multi-target antibacterial phytochemical, that exerts antagonism against aquaculture bacterial pathogens such as V. parahaemolytics and A. hydrophila.Communicated by Ramaswamy H. Sarma.

Stearyl palmitate a multi-target inhibitor against breast cancer: in-silico, in-vitro & in-vivo approach.

Multi-target inhibitors are currently trending in the pharmaceutical research, as they possess increased efficacy and reduced toxicity. In this study multi-target inhibitors for breast cancer are explored from a curated list of natural products, i.e. 4,670 phytochemicals belonging to 360 medicinal plants. In-silico screening of phytochemicals using SeeSAR and AutoDock Vina resulted in identification of Stearyl Palmitate as a potential drug molecule that inhibits three drug targets, i.e. HER-2, MEK-1 and PARP-1 proteins. Molecular Dynamics Simulation for 100 ns each for these three protein-ligand complexes using Desmond, Maestro platform also confirmed the prediction of multi-target inhibition by Stearyl Palmitate. Further in-vitro MTT assay demonstrated that Stearyl Palmitate has a significant IC50 value of 40 µM against MCF-7 cells and >1000 µM against L929 cells. This confirmed that Stearyl Palmitate is having selective cytotoxicity towards breast cancer cells in comparison to non-cancerous cells. Fluorescence staining and flow cytometry analysis confirmed that, Stearyl Palmitate is inducing apoptosis in MCF-7 cells at IC50 concentration. Finally, in-vivo efficacy and toxicity studies were performed using zebrafishes (Danio rerio). It was observed that the fishes treated with IC50 concentration of Stearyl Palmitate demonstrated 2x folds reduction in tumour size, while double dose resulted in 4x folds reduction in tumour size. Stearyl Palmitate did not demonstrate any toxicity or side effects in the zebrafishes. It is concluded that, Stearyl Palmitate, a phytochemical reported to be present in Althea officinalis is a potential anti-breast cancer agent, with ability to inhibit multiple targets such as HER-2, MEK-1 and PARP-2 proteins.Communicated by Ramaswamy H. Sarma.