The present study's objective was to identify the phytoconstituentspresent in n-hexane extract of Cissusquadrangularis(C.quadrangularis)with a help of Gas chromatography-Mass Spectrometry (GC-MS). The extract was subjected for the antibacterial (Escherichia coli ) and anthelmintic (adult Indian earthworms-Pheretimaposthuma)in-vitro activities followed by the validation of the same by subjecting the bioactive compounds obtained in GCMS analysis to molecular docking studies for anthelmintic and antibacterial protein targets.Threecompounds with molecular formula C20H42O2S, C14H25O3N2Cl and C30H50O were identified in GCMS analysis. Different extract concentrations viz 25, 50, 75 and 100 mg/ml and25, 50, 75 and 100μg/mlfor anthelmintic activity and antibacterial activityrespectively. For anthelmintic protein target, results of molecular docking (presented by 3D & 2D model) revealed that the identified compounds (binding energy -6.05 to -8.40 Kcal/mol) exhibited a comparable binding affinity as that of albendazole(binding energy -9.54 Kcal/mol) except C3 (binding energy +33.42 Kcal/mol) compound. Antibacterial protein target, compounds' binding affinities were found to be in the range -4.49 to -6.68 Kcal/mol, while Chlorobiocin was found to be -4.75 Kcal/mol. The extract exhibited dose-dependent in-vitroantihelmintic (75 and 100 mg/ml) and antibacterial activity (75 and 100 μg/ml). The minimum lethal amount of extract concentration for anthelmintic activity was found to be 100 μg/ml. At this concentration, Pheretimaposthumadied in31.09 ± 1.25min while with a standard dose of albendazole, the lethal time was found to be 38.54 ± 1.54 min. Finally,concluded that the identified bioactive compounds from C.quadrangularishave the potentialantihelmintic and antibacterial activity as justified by in-vitro biological evaluations and molecular docking studies.