Mimosa pudica Linn is a well-known perennial herb and is traditionally used in ayurvedic medicine for the treatment of various illnesses. Despite its abundance in nature, the therapeutic potential of this invasive weed is deemed to be underappreciated in Malaysia. Previous studies have found an abundance of bioactive compounds associated with potent antioxidant properties in all parts of the plant. However, the optimum parameters required for the extraction of antioxidant compounds are still unknown. Therefore, the present study aimed to optimize the solvent extraction parameters of M. pudica using response surface methodology to enrich the accumulation of antioxidant compounds in the extracts. The effects of the optimized M. pudica extracts were then evaluated on the cell viability and glucose uptake ability in a 3T3-L1 adipocyte cell line. The highest total phenolic (91.98 mg of gallic acid equivalent per g of the dry extract) and total flavonoid content (606.31 mg of quercetin equivalent per g of the dry extract) were recorded when using 100% ethanol that was five-fold and three-fold higher, respectively, as compared to using 50% ethanol. The extract concentration required to achieve 50% of antioxidant activity (IC50 value) was 42.0 µg/mL using 100% ethanol as compared to 975.03 µg/mL using 50% ethanol. The results indicated that the use of 100% ethanol solvent had the greatest impact on the accumulation of antioxidant compounds in the extract (p < 0.05). Cell viability assay revealed that all extract concentration treatments recorded a viability level of above 50%. Glucose uptake assay using 2-NBDG analog showed that the cells treated with 50 µg/mL extract combined with insulin were five-fold higher than the control group. Given the high antioxidant and antidiabetic properties of this plant, M. pudica can be easily highlighted as a plant subject of interest, which warrants further investigation for nutraceutical prospects.