Neem (Azadirachta indica [AI]) is a unique and traditional source of antioxidant and antibacterial compounds. The GC-MS studies revealed that phytoextract of Azadirachta indica comprises a large number of phytocompounds that possess the efficacy of inhibiting the biofilm. It was observed that phytocompounds like catechin showed maximum eradication of biofilm along with the degradation of EPS structural components like carbohydrates and proteins compared to quercetin, nimbolide, nimbin, and azardirachtin, and hence, catechin was proved to be the best against dental plaque-forming bacteria. It was also observed that catechin was able to bring about a marked reduction in quorum sensing (QS) both in Alcaligenes faecalis and Pseudomonas gingivalis dental biofilm-forming strains. The extent of such reduction was maximum for catechin (94.56±2.56% in P. gingivalis & 96.56±2.5 in A. faecalis) in comparison to other bioactive compounds. It was further observed that the bioactive compounds possess the ability to quickly pass across the membrane and bring about inhibition in the DNA and RNA content of the sessile cells. This was further validated by microscopic and in silico studies. Thus, this study revealed that catechin obtained from the phytoextract of AI showed a marked ability to inhibit the dental biofilm and can be used as a natural drug-like compound in treating biofilm-associated chronic infections.