<p>The rapidly growing antimicrobial resistance (AMR) in pathogenic bacteria has become one of the most critical public health challenges, with high mortality; hence, the development of innovative antibacterial agents and therapeutic techniques is urgently needed. In this study, an adenosine–benzoic acid conjugate containing both ester and amide functional moieties was synthesized with a yield of 32%, and its structure was confirmed by proton nuclear magnetic resonance (¹H NMR), carbon-13 nuclear magnetic resonance (¹³C NMR), Fourier-transform infrared (FT-IR) spectroscopy, and mass spectrometry (MS), where a prominent peak at m/z 476.09 [M + H]⁺ confirmed the target dibenzoylated structure. The synthesized conjugate was evaluated for its in vitro antibacterial and antibiofilm activity against Gram-positive <i>Streptococcus mutans</i> (<i>S. mutans</i>) and Gram-negative <i>Escherichia coli (E. coli)</i> bacteria, exhibiting a minimum inhibitory concentration (MIC) value of 1024&#xa0;µg/mL against both strains. The conjugate exhibited a strain-dependent antibiofilm profile; specifically, it outperformed free adenosine against <i>S. mutans</i>, whereas free benzoic acid demonstrated more potent inhibition against <i>E. coli</i>, achieving 87.01% inhibition compared to 44.87% for the conjugate at a sub-MIC of 512&#xa0;µg/mL. Molecular docking results provided a molecular basis for the conjugate’s interaction with target proteins (PDB IDs: 4LFU and 4TQX), suggesting its potential as a quorum-sensing modulator. These findings highlight the conjugate’s potential as a scaffold for modulating bacterial communication pathways, though future research is needed to optimize its efficacy.</p>