Computation and Experimental Approaches to Uncover the Antidiabetic Potential of Insulin Leaf (Tithonia diversifolia) Extract Based on Flavonoid Constituents

Insulin leaves (Tithonia diversifolia) are traditionally used by the community to lower blood sugar, although this has not been scientifically proven. Tithonia diversifolia is known to contain flavonoids, which are responsible for the antidiabetic activity of these leaves. Therefore, this research was conducted to investigate the antidiabetic effects of T. diversifolia, specifically through its flavonoid content. Tithonia diversifolia was extracted with 90% methanol and analyzed for flavonoid content by Liquid Chromatography-Mass Spectrometry (LC-MS). To investigate the antidiabetic activity, the sample extract was tested for its effect on the in vitro amylase enzyme and inhibition of glycated haemoglobin. Moreover, advanced techniques such as in silico analysis and molecular docking are used to elucidate the molecular mechanism behind the antidiabetic effects of T. diversifolia. The results of our study revealed that plant extracts, particularly those rich in flavonoids such as luteolin, genistein, and hispidulin, are potent inhibitors of the α-amylase enzyme and the formation of glycated haemoglobin. The Inhibition Concentration 50% (IC₅₀) values of flavonoids were lower than those of glibenclamide, suggesting their superior efficacy in stabilizing blood sugar levels. Docking analysis further confirmed the strong interactions between flavonoids and key enzyme residues and haemoglobin, while Absorption, Distribution, Metabolism, Excretion, and Toxicity (ADMET) predictions indicated favourable absorption and distribution profiles. Both experimental and computational approaches show the importance of flavonoids in the antidiabetic activity of T. diversifolia extracts, highlighting their contribution to its effectiveness.