The chemical and biological profiling of the root extracts of Rubia tinctorum was performed. The activities of different extracts were determined considering the antidiabetic effect against type II diabetes mellitus together with anti-obesity and hepatoprotective effects and lipid profile. The methanolic extract of Rubia tinctorum exhibited significant results in decreasing body weight, improving lipid profile, normalizing hyperglycaemia, insulin resistance, hyperinsulinemia. Additionally, it showed enhancement of liver tissue structure and function. The methanolic extract, being the most significant one, was subjected to LC-HRMS analysis to determine its chemical constituents. Finally, the chemical constituents were evaluated by molecular docking study that was carried out to identify the interaction of a panel of 45 compounds in silico and to correlate the structures to their anti-diabetic activity. Among the tested compounds, 1-hydroxy-2-hydroxymethyl anthra-quinone and naringenin-7-O-glucoside showed the most potent activity as α-amylase inhibitors.
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