Neohesperidin, a flavanone glycoside resourced from Curcuma amada rhizome, holds the key to the remediation of diabetes mellitus: An integrated experimental and computational study

Curcuma spp. is renowned for its therapeutic properties and holds prominence in traditional Chinese medicine (TCM). Popularly, the plant is referred to as Ézhú (莪术) in China. Traditional Chinese medicine holds enormous potential in addressing several diseases, including Diabetes Mellitus (DM), with a multipronged approach. The present study evaluates the anti-diabetic potency of Curcuma amada Roxb. While C. amada is not explicitly cited in traditional Chinese medicine (TCM) texts, TCM integrates various Curcuma spp. with comparable properties for their potential health benefits. In-vitro α -amylase and α -glucosidase enzyme inhibition was performed on bioactivity-guided fractions of crude ethyl acetate C. amada rhizome extract to determine the most bioactive fraction. Furthermore, a 2-NBDG (2-(7-Nitro-2,1,3-benzoxadiazol-4-yl)- d -glucosamine) mediated glucose uptake assay was carried out through a flow cytometric approach to corroborate the anti-diabetic property of the lead fraction. GC-MS-based metabolomic profiling was considered to determine the possible bioactive phytochemicals in the most effective bioactive fraction. Extensive in-silico computational and bioinformatics studies, including molecular dynamic simulations, were performed to depict the possible mechanism behind this anti-diabetic efficacy. The comprehensive in-vitro enzyme inhibition screening of fractions identified fraction 43 as the most potent anti-diabetic fraction, corroborated by glucose uptake studies. An extensive in-silico molecular docking study against 50 target receptors associated with DM, with the 21 unique phytocompounds obtained through the GC–MS-based metabolomic profiling study, identified neohesperidin as the most potent bioactive phytochemical. Computational chemistry employing DFT (Density Functional Theory) analysis identified neohesperidin as the strongest chemically reactive compound species in the most bioactive fraction. The molecular dynamic study highlighted that neohesperidin possibly remediated diabetes through TNF α (Tumor Necrosis Factor α) and IGF1R (Insulin-like Growth Factor 1 Receptor) pathways, as represented by the stable dynamic simulations, RMSD (Root Mean Square Deviation), and δG (Gibbs Free Energy) values. The in-vitro enzyme inhibition, glucose uptake, in-silico computational and bioinformatics studies reveal that neohesperidin resourced from Curcuma amada Roxb. has enormous potential as a future anti-diabetic drug.