Article
Author: Kofoed, Eric M. ; Clark, Kevin ; Liang, Xiaorong ; Yen, Chun-Wan ; Harris, Seth F. ; Hoag, Bridget ; Liu, Zhiguo ; Wu, Ping ; Ma, Fang ; Chen, Jacob ; Liang, Jun ; Mao, Jialin ; Zhang, Hua ; Crawford, Terry ; Lorenzo, Maria ; Reeder, Janina ; Aliagas, Ignacio ; Xu, Yiming ; Skelton, Nicholas ; Chen, Liuxi ; Liu, Xingrong ; Xu, Min ; Sims, Jessica ; Wang, Shumei ; Yan, Zhengyin ; Li, Chun Sing ; Yang, Ying ; Ubhayakar, Savita ; Peng, Yutian ; Tan, Man Wah ; Wai, John ; Park, Summer ; Skippington, Elizabeth ; Baumgardner, Matt
ABSTRACT
Guanine nucleotides are required for growth and viability of cells due to their structural role in DNA and RNA, and their regulatory roles in translation, signal transduction, and cell division. The natural antibiotic mycophenolic acid (MPA) targets the rate-limiting step in
de novo
guanine nucleotide biosynthesis executed by inosine-5´-monophosphate dehydrogenase (IMPDH). MPA is used clinically as an immunosuppressant, but whether
in vivo
inhibition of bacterial IMPDH (GuaB) is a valid antibacterial strategy is controversial. Here, we describe the discovery of extremely potent small molecule GuaB inhibitors (GuaBi) specific to pathogenic bacteria with a low frequency of on-target spontaneous resistance and bactericidal efficacy
in vivo
against
Acinetobacter baumannii
mouse models of infection. The spectrum of GuaBi activity includes multidrug-resistant pathogens that are a critical priority of new antibiotic development. Co-crystal structures of
A. baumannii, Staphylococcus aureus
, and
Escherichia coli
GuaB proteins bound to inhibitors show comparable binding modes of GuaBi across species and identifies key binding site residues that are predictive of whole-cell activity across both Gram-positive and Gram-negative clades of Bacteria. The clear
in vivo
efficacy of these small molecule GuaB inhibitors in a model of
A. baumannii
infection validates GuaB as an essential antibiotic target.
IMPORTANCE
The emergence of multidrug-resistant bacteria worldwide has renewed interest in discovering antibiotics with novel mechanism of action. For the first time ever, we demonstrate that pharmacological inhibition of
de novo
guanine biosynthesis is bactericidal in a mouse model of
Acinetobacter baumannii
infection. Structural analyses of novel inhibitors explain differences in biochemical and whole-cell activity across bacterial clades and underscore why this discovery may have broad translational impact on treatment of the most recalcitrant bacterial infections.