Assessing Spectral Analysis of Phytoconstituents and Their In Silico Interactions with Target Proteins in Plant Seed Extracts

Abstract

The pharmacological and preventive attributes of extracts from vegetable seeds have garnered widespread recognition within the scientific community. This study systematically assessed the in vitro antibacterial, antioxidant, and anti-breast cancer properties of phytochemicals present in various solvent-based vegetable seed extracts. We also conducted molecular docking simulations to ascertain their interactions with specific target proteins. Besides, nine distinct chemical constituents were identified using gas chromatography-mass spectrometry (GCMS). Remarkably, the ethyl acetate extract exhibited robust inhibitory effects against Gram-positive and Gram-negative bacterial strains. Furthermore, its capacity for 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging was found to be noteworthy, with an IC₅₀ value of 550.82 ± 1.7 µg/mL, representing a scavenging efficiency of 64.1 ± 2.8%. Additionally, the ethyl acetate extract demonstrated significant hydrogen peroxide (H₂O₂) scavenging activity, with a maximal scavenging rate of 44.1 ± 1.70% (IC₅₀) at a concentration of 761.17 ± 1.8 µg/mL. Intriguingly, in vitro cytotoxicity assays against human breast cancer (MCF-7) cells revealed varying levels of cell viability at different extract concentrations, suggesting potential anticancer properties. Importantly, these ethyl acetate extracts did not display toxicity to L929 cells across the concentration range tested. Subsequently, we conducted in-silico molecular docking experiments utilizing Discovery Studio 4.0 against the c-Met kinase protein (hepatocyte growth factor; PDB ID: 1N0W). Among the various compounds assessed, 3,4-Dihydroxy-1,6-bis-(3-methoxy-phenyl)-hexa-2,4-diene-1,6-dione exhibited a notable binding energy of -9.1 kcal/mol, warranting further investigation into its potential anticancer properties, clinical applications, and broader pharmacological characteristics.

Keywords: GC-MS; anti-breast cancer; antibacterial; antioxidant; in-silico molecular docking; vegetable seed.

Figure 1

Antioxidant activity of vegetable seed extracts of hexane, ethyl acetate, methanol extracts and ascorbic acid. (a) DPPH scavenging effect of different concentrations (200–1000 µg/mL). (b) H₂O₂ scavenging effect of different concentrations (200–1000 µg/mL) of vegetable seed extracts of hexane, ethyl acetate, methanol extracts, and ascorbic acid. Each value represents the mean ± SEM of triplicate experiments.

Figure 2

Gas chromatography–mass spectrometry chromatograms of the ethyl acetate extract of vegetable seed (Synergistic).

Figure 3

Mass spectrum for the major constituent methyl oleate from the ethyl acetate extract of vegetable seed (Synergistic).

Figure 4

Anticancer activity of L-929 (Fibroblast cells) ethyl acetate extracts (a) control, (b) 500 µg/ mL, (c) 250 µg/mL, (d) 100 µg/mL, (e) 50 µg/mL, and (f) 25 µg/mL.

Figure 5

Anticancer activity of ethyl acetate extracts on MCF-7 (a) control, (b) 25 µg/mL, (c) 50 µg/mL, (d) 100 µg/mL, (e) 250 µg/mL, and (f) 500 µg/ mL.

Figure 6

Anticancer activities of ethyl acetate extract from vegetable seed against MCF-7 and L929 cell line.

Figure 7

(a) Ligand–receptor hydrogen bond interactions of 3,4-Dihydroxy-1,6-bis-(3-methoxy-phenyl)-hexa-2,4-diene-1,6-dione. (b) Receptor side hydrogen bond interactions of3,4-Dihydroxy-1,6-bis-(3-methoxy-phenyl)-hexa-2,4-diene-1,6-dione. (c) 2D-ligand–receptor hydrogen bond interaction of 3,4-Dihydroxy-1,6-bis-(3-methoxy-phenyl)-hexa-2,4-diene-1,6-dione. (d) Ligand–receptor hydrogen bond interactions of Phenol,2,6-bis(1,1-dimethylethyl)-4-(4-hydroxy-3,5-dimethylphenyl)methyl). (e) Receptor side hydrogen bond interactions of Phenol,2,6-bis(1,1-dimethylethyl)-4-(4-hydroxy-3,5-dimethylphenyl)methyl). (f) 2D-ligand–receptor hydrogen bond interaction of Phenol,2,6-bis(1,1-dimethylethyl)-4-(4-hydroxy-3,5-dimethylphenyl)methyl). (g) Ligand–receptor hydrogen bond interactions of Isopropyl stearate. (h) Receptor side hydrogen bond interactions of Isopropyl stearate. (i) 2D-ligand–receptor hydrogen bond interaction of Isopropyl stearate.