<p>We have synthesized and generated a range of bioactive dispersed dyes that may be joined with pyrimidine moieties inside the same construct via an azo linker, thus taking advantage of both compounds’ bioactive properties. Through a straightforward process, the heterocyclic azo-disperse dyes <b>3a–l</b> were produced by diazo coupling aryldiazonium chloride derivatives with 2-(4-hydroxyphenyl)amino-6-oxo-4-phenyl-1,6-dihydropyrimidine-5-carbonitrile <b>(2)</b> in good yields. Mass spectroscopy, elemental analysis, FT-IR, ¹H-NMR, and ¹³C-NMR characterized and confirmed the newly synthesized compounds’ structures. The color characteristics of the dyes and their fastness properties, such as resistance to washing, perspiration, light, and rubbing, were evaluated. The antibacterial efficacy of the printed fabrics was assessed using the colony-forming unit (CFU) method. The bioassay outcomes showed that some targeted compounds have moderate to excellent antibacterial activity (in vitro). The activity of the synthesized azo dye molecules and their dyes-loaded polyester fabrics was evaluated against clinically important bacterial pathogens. According to our data, the electron-withdrawing groups in compounds <b>3b</b>, <b>3d</b>, <b>3i</b>, and <b>3&#xa0;k</b> showed comparatively great activity with significantly lower MIC values. Furthermore, the printed sample infused with azo dyes demonstrated an inhibitory efficacy against all bacterial infections, indicating their potential for use in textiles intended in medical materials.</p> Graphical abstract <p></p>