The objective of the study was to evaluate the anti-biofilm activity of Indian medicinal plant Tylophora indica extracts against the biofilm forming Streptococcus pyogenes isolated from pharyngitis patients. Methods: The plant extracts (methanol, ethyl acetate, chloroform and petroleum ether) were screened for their preliminary phytochemical components. The solvent extract with higher phytochemical yield was subjected to quantitative analysis using the Gas Chromatography-Mass Spectrometry (GC-MS) technique. In vitro analysis of the anti-biofilm study was performed using the Minimal Inhibitory Concentration (MIC) assay, biofilm inhibitory concentration assay, growth curve analysis, anti-bacterial activity, and light microscopy analysis. Results: The methanol extract exhibited the strong phytochemical components. GC-MS analysis of the methanol extract showed a total of thirty seven phyto compounds among which most of the compounds were medicinally important. MIC assay showed that the inhibition of test pathogens was at an average concentration of 2 mg/ml. whereas the biofilms of the test pathogens were significantly reduced and were dose dependent at sub-MIC levels as confirmed by the light microscopic analysis. Conclusion: These preliminary results indicated that the methanol extract of T. indica leaves consisted of pharmacologically active components and could be used as an anti-biofilm agent at minimal concentrations thereby successfully preventing the formation of biofilms.