The alarming rise of bacterial antibiotic resistance requires the development of new compounds. Such compounds, lipophosphonoxins (LPPOs), were previously reported to be active against numerous bacterial species, but serum albumins abolished their activity. Here we describe the synthesis and evaluation of novel antibacterial compounds termed LEGO-LPPOs, loosely based on LPPOs, consisting of a central linker module with two attached connector modules on either side. The connector modules are then decorated with polar and hydrophobic modules. We performed an extensive structure-activity relationship study by varying the length of the linker and hydrophobic modules. The best compounds were active against both Gram-negative and Gram-positive species including multiresistant strains and persisters. LEGO-LPPOs act by first depleting the membrane potential and then creating pores in the cytoplasmic membrane. Importantly, their efficacy is not affected by the presence of serum albumins. Low cytotoxicity and low propensity for resistance development demonstrate their potential for therapeutic use.

Department of Analytical Chemistry Faculty of Science Charles University Albertov 6 128 43 Prague 2 Czech Republic

Department of Genetics and Microbiology Faculty of Science Charles University Viničná 5 128 43 Prague 2 Czech Republic

Department of Medical Chemistry and Biochemistry Faculty of Medicine and Dentistry Palacký University Olomouc Hněvotínská 3 775 15 Olomouc Czech Republic

Department of Microbiology Faculty of Medicine and Dentistry Palacký University Olomouc Hněvotínská 3 775 15 Olomouc Czech Republic

Institute of Microbiology Czech Academy of Sciences v v i Vídečská 1083 142 20 Prague 4 Czech Republic

Institute of Organic Chemistry and Biochemistry Czech Academy of Sciences v v i Flemingovo nám 2 166 10 Prague 6 Czech Republic

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