Antibiotic resistance caused by biofilm formation is a clinical challenge. Nitric oxide (NO) can effectively disperse a mature biofilm and can also synergistically influence the level of cyclic diguanylate (c-di-GMP), a universal secondary messenger that plays an important role in biofilm formation in bacteria. Based on our previous finding that c-di-GMP G-quadruplex inducers are effective biofilm formation inhibitors, we designed and synthesized a c-di-GMP G-quadruplex inducer-NO donor conjugate () as a bifunctional antibiofilm agent after obtaining the c-di-GMP G-quadruplex inducer (), which has an amino group capable of binding to a nitroso group (NO donor). The conjugate showed better biofilm inhibition efficiency than , and it can also eradicate mature biofilm. Additionally, it exhibited good antimicrobial synergism against and helped elevate the bactericidal efficiency of tobramycin against biofilm-formed bacteria. In combination with tobramycin, also improved the survival rate of in a hyperbiofilm environment.
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