In Vitro, In Silico and Network Pharmacology Mechanistic Approach to Investigate the a-Glucosidase Inhibitors Identified by Q-ToF-LCMS from Phaleria macrocarpa Fruit Subcritical CO2 Extract

The fruit of Phaleria macrocarpa have been traditionally used as an antidiabetic remedy in Malaysia and neighbouring countries. Despite its potential for diabetes treatment, no scientific study has ever been conducted to predict the inhibitor interaction of the protein alpha-glucosidase identified in an extract prepared with a non-conventional extraction technique. Hence, the major aim of this research was to evaluate the in vitro antioxidant, the alpha-glucosidase inhibitors, and the molecular dynamic simulations of the alpha-glucosidase inhibitors identified by Quadrupole Time-of-Flight Liquid Chromatography Mass Spectrometry (Q-ToF-LCMS) analysis. Initially, dry fruit were processed using non-conventional and conventional extraction methods to obtain subcritical carbon dioxide extracts (SCE-1 and SCE-2) and heating under reflux extract (HRE), respectively. Subsequently, all extracts were evaluated for their in vitro antioxidative and alpha-glucosidase inhibitory potentials. Subsequently, the most bioactive extract (SCE-2) was subjected to Q-ToF-LCMS analysis to confirm the presence of alpha-glucosidase inhibitors, which were then analysed through molecular dynamic simulations and network pharmacology approaches to confirm their possible mechanism of action. The highest inhibitory effects of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and alpha-glucosidase on SCE-2 was found as 75.36 +/- 0.82% and 81.79 +/- 0.82%, respectively, compared to the SCE-1 and HRE samples. The Q-ToF-LCMS analysis tentatively identified 14 potent alpha-glucosidase inhibitors. Finally, five identified compounds, viz., lupenone, swertianolin, m-coumaric acid, pantothenic acid, and 8-C-glucopyranosyleriodictylol displayed significant stability, compactness, stronger protein-ligand interaction up to 100 ns further confirming their potential as alpha-glucosidase inhibitors. Consequently, it was concluded that the SCE-2 possesses a strong alpha-glucosidase inhibitory effect due to the presence of these compounds. The findings of this study might prove useful to develop these compounds as alternative safe alpha-glucosidase inhibitors to manage diabetes more effectively.

Automated summary

This study evaluated the potential therapeutic effects of Phaleria macrocarpa fruit extracts on diabetes, specifically focusing on their antioxidant properties and their ability to inhibit the protein alpha-glucosidase. The study confirmed the presence of potent alpha-glucosidase inhibitors in the extracts and analyzed their mechanisms of action using molecular dynamic simulations and network pharmacology approaches. The findings suggest that these compounds have the potential to be developed as alternative alpha-glucosidase inhibitors for more effective diabetes management.