The present work involves synthesis of 1-substitutedindole-2,3dione-2-thiosemicarbazone derivatives in order to evaluate their antimicrobial, anti-inflammatory, analgesic, and anticonvulsant activities. The target compounds were prepared through oxidative deacetylation of 1-substituted-2-acetyl-3-hydroxyindole, which followed by the condensation with different thiosemicarbazide derivatives. The purity of all the newly synthesized compounds was checked by TLC and elucidation of their structures was confirmed by IR, 1H-NMR, and some representatives by mass spectrometry along with elemental microanalyses. Preliminary in-vitro antimicrobial evaluation (MIC) against some gram-positive and gram-negative bacteria as well as some fungi revealed that the tested compounds showed variable degrees of antibacterial activity, with little antifungal activity in comparison to chloramphenicol and fluconazole as reference drugs respectively. In addition, some of the tested compounds showed anti-inflammatory activity comparable to that of indomethacin. The most active compounds were further evaluated for their analgesic activity; results showed that these compounds were more active than indomethacin. Three compounds were tested for their ulcerogenicity; two of them were safer than indomethacin. Furthermore, in-vivo anticonvulsant evaluation of the tested compounds at 2.8, 2.0 and 1.4 mmol/Kg concentrations showed comparable anticonvulsant activity to that of diazepam using pentylenetetrazole induced seizure protocol. Moreover, LD50 of the most active compounds 4b and 4h were found to be 250 and 300 mg/Kg; respectively in comparison to the reported one for indomethacin 13 mg/Kg (i.p.). Moreover, the study involved the docking of the most active compound in the active site of COX-2 enzyme.