The present study aims to explore the therapeutic efficacy of a polyherbal formulation derived from plants indigenous to Kandhamal District, Odisha, with potent anti-inflammatory, anti-microbial, and hepatoprotective properties. The formulation was developed and standardized using a combination of Cayratia trifolia, Sesbania grandiflora, Cordia dichotoma, and Teprosia purpurea, known for their traditional medicinal uses. The polyherbal formulation was subjected to rigorous evaluation to assess its antiinflammatory activity. In vitro assays revealed significant inhibition of TNF-α, IL-6, and COX-2, indicating its potential in modulating inflammation. Furthermore, the formulation exhibited remarkable anti-microbial activity against pathogens such as E. coli, S. aureus, and C. albicans, as evident from the inhibition zones observed. Hepatoprotective activity was also assessed, and the results demonstrated the formulation's ability to mitigate liver damage. Various parameters, including ALT, AST, LDH, ROS levels, MDA levels, SOD activity, and GSH levels, were measured, and the formulation showed promising hepatoprotective effects by maintaining their homeostasis. To further understand the composition of the polyherbal formulation, phytochemical analysis was performed. The presence of alkaloids, proteins and amino acids, carbohydrates, flavonoids, tannins, and saponins was confirmed, indicating the richness of bioactive compounds in the formulation. TLC analysis using a specific solvent system exhibited distinct spots for the marker compounds, gallic acid, and piperine, validating their presence in the formulation. Quantification of total phenolic and flavonoid content revealed substantial amounts of these compounds in the formulation, further supporting its antioxidant potential. The presence of polyphenolics and flavonoids contributes to the rationality behind the formulation's observed antioxidant activity. So, the developed polyherbal formulation from plants collected from Kandhamal District, Odisha, and exhibits significant therapeutic efficacy against inflammation, microbial infections, and liver damage. The comprehensive analysis of its phytochemical composition and validation of its antioxidant properties highlight its potential as a natural remedy for various health ailments.