<jats:title>Abstract</jats:title><jats:p>Endophytic fungi, especially from mangrove plants, are rich source of secondary metabolites, which plays a major role in various pharmacological actions preferably in cancer and bacterial infections. To perceive its role in antidiabetic activity we isolated and tested the metabolites derived from a novel strain <jats:italic>Alternaria longipes</jats:italic> strain VITN14G obtained from mangrove plant <jats:italic>Avicennia officinalis</jats:italic>. The crude extract was analyzed for antidiabetic activity and subjected to column chromatography. The isolated fractions were screened in vitro for α‐glucosidase and α‐amylase inhibitory activities. The cytotoxicity of the isolated fractions was studied on L929 cell lines. Following which, the screened fraction 2 was allowed for structure elucidation using gas chromatography‐mass spectrometry, one‐dimensional, two‐dimensional nuclear magnetic resonance spectroscopy, ultraviolet, and Fourier‐transform infrared analysis. The binding energies of the isolated fraction 2 with glycolytic enzymes were calculated by molecular docking studies using AutoDock Vina. The isolated fraction 2 identified as 2,4,6‐triphenylaniline, showed no significant difference in α‐amylase inhibition rates and a significant difference of 10% in α‐glucosidase inhibition rates than that of the standard drug acarbose. Further, the cytotoxicity assay of the isolated fraction 2 resulted in a cell viability of 73.96%. Supportingly, in silico studies showed 2,4,6‐triphenylaniline to produce a stronger binding affinity toward the glycolytic enzyme targets. The compound 2,4,6‐triphenylaniline isolated from <jats:italic>A. longipes</jats:italic> strain VITN14G exhibited satisfactory antidiabetic activity for type 2 diabetes in vitro, which will further be confirmed by in vivo studies. Successful outcome of the study will result in a natural substitute for existing synthetic antidiabetic drugs.