Cinnamaldehyde derivatives act as antimicrobial agents against Acinetobacter baumannii through the inhibition of cell division

Abstract

<i>Acinetobacter baumannii</i> is a pathogen with high intrinsic antimicrobial resistance while multidrug resistant (MDR) and extensively drug resistant (XDR) strains of this pathogen are emerging. Treatment options for infections by these strains are very limited, hence new therapies are urgently needed. The bacterial cell division protein, FtsZ, is a promising drug target for the development of novel antimicrobial agents. We have previously reported limited activity of cinnamaldehyde analogs against <i>Escherichia coli</i>. In this study, we have determined the antimicrobial activity of six cinnamaldehyde analogs for antimicrobial activity against <i>A. baumannii</i>. Microscopic analysis was performed to determine if the compounds inhibit cell division. The on-target effect of the compounds was assessed by analyzing their effect on polymerization and on the GTPase activity of purified FtsZ from <i>A. baumannii</i>. <i>In silico</i> docking was used to assess the binding of cinnamaldehyde analogs. Finally, <i>in vivo</i> and <i>in vitro</i> safety assays were performed. All six compounds displayed antibacterial activity against the critical priority pathogen <i>A. baumannii</i>, with 4-bromophenyl-substituted <b>4</b> displaying the most potent antimicrobial activity (MIC 32 μg/mL). Bioactivity was significantly increased in the presence of an efflux pump inhibitor for <i>A. baumannii</i> ATCC 19606 (up to 32-fold) and significantly, for extensively drug resistant UW 5075 (greater than 4-fold), suggesting that efflux contributes to the intrinsic resistance of <i>A. baumannii</i> against these agents. The compounds inhibited cell division in <i>A. baumannii</i> as observed by the elongated phenotype and targeted the FtsZ protein as seen from the inhibition of polymerization and GTPase activity. <i>In silico</i> docking predicted that the compounds bind in the interdomain cleft adjacent to the H7 core helix. Di-chlorinated <b>6</b> was devoid of hemolytic activity and cytotoxicity against mammalian cells <i>in vitro</i>, as well as adverse activity in a <i>Caenorhabditis elegans</i> nematode model <i>in vivo</i>. Together, these findings present halogenated analogs <b>4</b> and <b>6</b> as promising candidates for further development as antimicrobial agents aimed at combating <i>A. baumannii</i>. This is also the first report of FtsZ-targeting compounds with activity against an XDR <i>A. baumannii</i> strain.