Introduction: Corchorus olitorius L. is a significant plant in folk medicine, therefore, research on phytochemistry with biological activity may reveal its potential use in drug development. Medicinal plants are valuable sources of drug materials as they include bioactive compounds known as secondary metabolites. Determination of bioactive compounds and biological activity of plants lead to understanding their potential in drug development. Methods: A quantitative analysis of the bioactive compounds in leaf, stem, and root extracts of C. olitorius L. was carried out with Liquid Chromatography-Mass Spectrometry (LC-MS/MS). Acetylcholinesterase (AChE) and Glutathione-S-Transferase (GST) enzyme inhibition and antibacterial effects were investigated using Mueller Hinton broth microdilution assay. The anticancer activity of C. olitorius leaves was evaluated by MTT assay using HDF, U87, Skov-3, and Caco-2 cell lines. Results: In the quantitative study, leaf extract exhibited the largest concentration of bioactive compounds, and cynarin (6.680 mg/g extract) and chlorogenic acid (5.605 mg/g extract) were the major products. The leaf, stem, and root extracts showed significant AChE and GST inhibition activity. The leaf extract displayed antibacterial activity against P. aeruginosa with the value of 2.2 +/- 0.1 (IC50, mu g/ mL) in comparison to the standard (4.00 +/- 0.1 IC50, mu g/ mL). The leaf extract displayed good antiproliferative activity against sk-ov-3 cell lines with values of 37.1% and 42.1% at 250 mu g/mL and 500 mu g/mL concentrations, respectively. Conclusion: C. olitorius has the potential to be a drug agent against cancer and microbial-induced diseases due to its effective bioactive compound contents.