Synergistic Effects of Thiosemicarbazides with Clinical Drugs against <i>S. aureus</i>

• Main Authors: Beata Chudzik-Rząd, Anna Malm, Nazar Trotsko, Monika Wujec, Tomasz Plech, Agata Paneth
• Format: Article
• Language: English
• Published: MDPI AG 2020-05-01
• Series: Molecules
• Subjects: thiosemicarbazides; antibacterial activity; synergistic effect; bacterial topoisomerases
• Online Access: https://www.mdpi.com/1420-3049/25/10/2302

Antimicrobial resistance spurred by the overuse and misuse of antibiotics is a major global health concern, and of the Gram positive bacteria, <i>S. aureus</i> is a leading cause of mortality and morbidity. Alternative strategies to treat <i>S. aureus</i> infections, such as combination therapy, are urgently needed. In this study, a checkerboard method was used to evaluate synergistic interactions between nine thiosemicarbazides (4-benzoyl-1-(2,3-dichloro-benzoyl)thiosemicarbazides <b>1</b>–<b>5</b> and 4-aryl-1-(2-fluorobenzoyl)thiosemicarbazides <b>6</b>–<b>9</b>) and conventional antibiotics against <i>S. aureus</i> ATCC 25923, which were determined as the fractional inhibitory concentration indices (FICIs). For these experiments, amoxicillin, gentamicin, levofloxacin, linezolid, and vancomycin were selected to represent the five antimicrobial classes most commonly used in clinical practice. With one exception of <b>7</b>-vancomycin combination, none of the forty-five thiosemicarbazide-antibiotic combinations tested had an antagonistic effect, showing promising results with respect to a combination therapy. The synergic effect was observed for the <b>2</b>-linezolid, <b>4</b>-levofloxacin, <b>5</b>-linezolid, <b>6</b>-gentamicin, <b>6</b>-linezolid, and <b>7</b>-levofloxacin combinations. No interactions were seen in combination of the thiosemicarbazide with gentamicin or vancomycin, whereas all combinations with linezolid acted in additive or synergism, except for <b>6</b>-gentamicin and <b>7</b>-linezolid. The 4-(4-chlorophenyl)-1-(2-fluorobenzoyl)thiosemicarbazide <b>6</b> showed a clear preference for the potency; it affected synergistically in combinations with gentamicin or linezolid and additively in combinations with amoxicillin, levofloxacin, or vancomycin. In further studies, the inhibitory potency of the thiosemicarbazides against <i>S. aureus</i> DNA gyrase and topoisomerase IV was examined to clarify the molecular mechanism involved in their synergistic effect in combination with levofloxacin. The most potent synergist <b>6</b> at concentration of 100 µM was able to inhibit ~50% activity of <i>S. aureus</i> DNA gyrase, thereby suggesting that its anti-gyrase activity, although weak, may be a possible factor contributing to its synergism effect in combination with linezolid or gentamycin.