Synergistic antimicrobial activities of epigallocatechin gallate, myricetin, daidzein, gallic acid, epicatechin, 3-hydroxy-6-methoxyflavone and genistein combined with antibiotics against ESKAPE pathogens

Abstract

Aim: To verify synergistic effects, we investigated the antimicrobial activity of seven phenolic phytochemicals (gallic acid; epicatechin; epigallocatechin gallate; daidzein; genistein; myricetin; 3-hydroxy-6-methoxyflavone) in combination with six antibiotics against multidrug-resistant isolates from the ESKAPE group. Methods and Results: To investigate single phytochemicals and combinations, initial microdilution and checkerboard assays were used, followed by time-kill assays to evaluate the obtained results. The research revealed that phenolic compounds on their own resulted in little or no inhibitory effects. During preliminary tests, most of the combinations resulted in indifference (134 [71.3%]). In all, 30 combinations led to antagonism (15.9%); however, 24 showed synergistic effects (12.8%). The main tests resulted in nine synergistic combinations for the treatment of four different bacteria strains, including two substances (3-hydroxy-6-methoxyflavone, genistein) never tested before in such setup. Time-kill curves for combinations with possible synergistic effects confirmed the results against Acinetobacter baumannii as the one with the greatest need for research. Conclusions: The results highlight the potential use of antibiotic–phytocompound combinations for combating infections with multi-resistant pathogens. Synergistic combinations could downregulate the resistance mechanisms of bacteria. Significance and Impact of the Study: The aim of this study is to demonstrate the potential use of phenolic natural compounds in combination with conventional antibiotics against multidrug-resistant bacteria of the ESKAPE group. Due to synergistic effects of natural phenolic compounds combined with antibiotics, pathogens that are already resistant to antibiotics could be resensitized as we were able to reduce their MICs back to sensitive. In addition, combination therapies could prevent the development of resistance by reducing the dose of antibiotics. This approach opens up the basis for future development of antimicrobial therapy strategies, which are so urgently needed in the age of multidrug-resistant pathogens.