Diabetes mellitus is a chronic metabolic disorder characterized by elevated blood glucose levels due to impaired insulin production or insulin resistance. As a result, there is a growing interest in exploring natural alternatives for the treatment and prevention of diabetes. Gossypium herbaceum has been traditionally used in various traditional medicine systems, including Ayurveda, for its therapeutic benefits. Therefore, the aim of this study was to evaluate the in vitro antioxidant potentials, antidiabetic and neuroprotective properties of ethanolic extract of Gossypium herbaceum leaves as well as its HPLC-identified phytoconstituents. The results oh HPLC analysis showed the presence of thirteen bioactive compounds such as Gossypol, choline, catechin, epicatechin, gallocatechin, epigallocatechin, sitosterol, ergosterol, gossycaerulin, gossypupurin, flavan-3-ol, cyanidin, delphinidin with their heights and retention times. Also, according to the antioxidant potentials of ethanolic extract of Gossypium herbaceum, it revealed higher Total Phenolic Contents (TPC) than the Total Flavonoid Contents (TFC) as well as a significant (p<0.05) inhibitory effect against DPPH and ABTs in concentration dependent manner. In addition, ethanolic extract of Gossypium herbaceum demonstrated a significant (p<0.05) inhibitory effect against α-amylase and α-glucosidase in concentration-dependent manner. Similarly, the extract had a significant (p<0.05) inhibition against Acetylcholinesterase (AChE) and Butylrylcholinesterase (BChE) in concentration-dependent manner. Therefore, finding of this study suggest that ethanolic extract of Gossypium herbaceum could probably have demonstrated antidiabetic and neuroprotective effects owing to the presence of bioactive phytonutrients as revealed by the HPLC technique. Furthermore, this property could be explored in the management of these afore-mentioned human diseases and beneficial pharmaceutical products.