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An imbalance of homeostasis between the microbial communities and the host system leads to dysbiosis in oral micro flora. DMTU (1,3-di-m-tolyl-urea) is a biocompatible compound that was shown to inhibit <i>Streptococcus mutans</i> biofilm by inhibiting its communication system (quorum sensing). Here, we hypothesized that DMTU is able to inhibit multispecies biofilms. We developed a multispecies oral biofilm model, comprising an early colonizer <i>Streptococcus gordonii</i>, a bridge colonizer <i>Fusobacterium nucleatum</i>, and late colonizers <i>Porphyromonas gingivalis</i> and <i>Aggregatibacter actinomycetemcomitans.</i> We performed comprehensive investigations to demonstrate the effect of DMTU on planktonic cells and biofilms. Our findings showed that DMTU inhibits and disrupts multispecies biofilms without bactericidal effects. Mechanistic studies revealed a significant down regulation of biofilm and virulence-related genes in <i>P. gingivalis</i>. Taken together, our study highlights the potential of DMTU to inhibit polymicrobial biofilm communities and their virulence.