The current study emphasizes on the utilization of the invasive allelopathic plant, Wedelia trilobata for its medicinal effect. It provides an elaborative phytochemical analysis of acetone extract (NALTW-1) and its hexane fraction (NALTW-H1) of Wedelia trilobata leaves, prepared by accelerated solvent extraction technique. The samples (NALTW-1 and NALTW-H1) were evaluated for their antioxidant potential in terms of2, 2´-azinobis[3-ethylbenzthiazoline]-6-sulfonic acid (ABTS) and 2,2- diphenyl-1-picrylhydrazyl(DPPH) assays and were also subjected to qualitative phytochemical analysis. They were characterized by high performance liquid chromatography (HPTLC) for quantifying antioxidant compounds (ascorbic acid, gallic acid, rutin and quercetin) and nucleobases (adenine, cytosine, guanosine, thymine and uracil). The results of the phytochemical analyses: ABTS and DPPH assays revealed substantial antioxidant potential of the samples. Considering these results, the samples were further evaluated for their anti-proliferative activity against the breast carcinoma cell line, MCF-7 using 3-(4,5-dimethylthiazol-2- yl)-2,5- diphenyltetrazolium (MTT) assay. In vitro, NALTW-1 and NALTW-H1 demonstrated moderate anti-proliferative activity against MCF-7 cells with IC50 values 570.78 μg/ml and 44.50.98 μg/ml, respectively. Thus, this study highlighted therapeutic benefit ofthe invasive plant, Wedelia trilobata.