Abstract
Objective: This investigation includes characterization of phytochemicals from acetone extract of Helianthus annuus L. seeds responsible for α-amylase and α-glucosidase inhibition revealed from in vitro and in silico approaches.Methods: Seed extract was qualitatively and quantitatively analysed for the presence of bioactive molecules. In vitro α-amylase and α-glucosidase inhibition assays and kinetics studies for α-glucosidase were done. Thin layer chromatography (TLC) autography of extract was done to screen potent inhibitors and characterized by high-resolution liquid chromatography-mass spectrometry (HR LC-MS). Characterized molecules were further used for in silico studies.Results: Qualitative investigation reveals the presence of flavonoids, glycosides, alkaloids, terpenoids, and steroids. Quantitative analysis for total phenolic content and total flavonoid content of the extract was 0.1±0.005 mg/ml GAE and 0.025±0.003 mg/ml QE respectively. Percent inhibition of α-amylase and α-glucosidase ascertained in presence of extract was 60.42±0.6 and 83.22±0.18 at 0.01 mg while 36.24±0.81 and 37.67±0.15 at 0.005 mg of extracts for both enzymes respectively. Kinetics studies of α-glucosidase inhibition illustrated the non-competitive type of inhibition. TLC autography inhibition patterns were characterized by HR LC-MS. Characterized molecules on docking revealed (6RS)-22-hydroxy-23,24,25,26,27-pentanor-vitamin-D3-6,19-sulfurdioxide-adduct, manoalide and 5β-cholestane-3α,7α,12α,24,25,26-hexol as the best docked molecules with lowest binding energies of-12.5,-11 and-10.2 kcal/mol for α-amylase and-14.2,-11 and-11.2 kcal/mol for α-glucosidase respectively.Conclusion: Results clearly suggested that (6RS)-22-hydroxy-23,24,25,26,27-pentanor-vitamin-D3-6,19-sulfurdioxide-adduct, manoalide and 5β-cholestane-3α,7α,12α,24,25,26-hexol could be considered as lead molecules for the discovery of potent antidiabetic agents.
Highlights
Helianthus annuus L. (Sunflower) is an important oilseed crop of the world at the third position in production next to groundnut and soybean [1, 2]
The total phenolic content of the extract was determined by FolinCiocalteu reagent method and measured in terms of gallic acid equivalent (GAE) showed 0.1 mg/ml concentration whereas, the total flavonoid content measured in terms of QE showed 0.025 mg/ml concentration as depicted in the table 2
Screening of phytoconstituents as α-amylase and α-glucosidase inhibitors with low or no side effects was the primary aim of this investigation
Summary
Helianthus annuus L. (Sunflower) is an important oilseed crop of the world at the third position in production next to groundnut and soybean [1, 2]. (Sunflower) is an important oilseed crop of the world at the third position in production next to groundnut and soybean [1, 2]. The importance of Helianthus annuus L. as a source of edible oil as well as high-quality protein is continuously increasing [3]. In addition to its nutritional benefits, sunflower seeds are reported to have pharmacological activities which may be the result of the phytochemicals they possess. Apart from these, phytochemicals like flavonoids, tannins, alkaloids, saponins, phytosterols, steroids and fixed oils were reported in methanolic extracts of seeds of Helianthus annuus L. Clorogenic acid and dicaffeoylquinic acid isolated from aqueous methanol extracts of sunflower are an addition to the list [5]
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