Studies proposed in this R21 application address the urgent need to develop new classes of antibiotics against emerging infectious agents, as well as pathogenic bacteria with the potential to be used as bio-weapons. The long-term goal of the work proposed in this application is to develop a new class of inhibitors against glutamate racemases, which catalyze the stereo-inversion of L- to D-glutamate, an important metabolite for cell wall biosynthesis. Glutamate racemases are essential in several bacteria, but not found in mammals, and are thus predicted to be excellent antibiotic targets. Notably, at least three pharmaceutical companies are currently developing glutamate racemase inhibitors as potential antimicrobial drugs, thereby supporting the potential importance of the glutamate racemases as antimicrobial targets. However, our strategy for inhibitor design is entirely different than these companies, and is based on the transition state structure of glutamate racemase, which we predict will bind to the enzyme with higher affinity than do drugs based on the ground state enzyme-substrate complexes. In this R21 application, we propose exploratory studies towards the goals of (i) characterizing the importance and properties of the two Bacillus anthracis glutamate racemases, RacE1 and RacE2, in vitro and in vivo, and, (ii) generating models of the transition state structures of the reactions catalyzed by both enzymes. This highly interdisciplinary application consolidates considerable expertise in bioorganic and computational chemistry, biochemistry, and bacterial pathogenesis.
The Specific Aims are:
Specific Aim 1. To characterize the importance and roles of racE1 and racE2.
Specific Aim 2. To characterize the transition states of RacE1- and RacE2-catalyzed racemization. The anticipated outcomes of these specific aims will be validation of glutamate racemase as a drug-target in B. anthracis, and the generation of transition state models for both RacE1 and RacE2. From these models, we will identify small molecule transition state analogs that will be screened for inhibitory activities against RacE1 and/or RacE2 enzyme activities. The results from these studies will provide the experimental and conceptual framework for future work to optimize small molecule """"""""leads"""""""" into ultra-specific, reaction-based inhibitors with antimicrobial activity.
This application addressed an existing and urgent need to develop new classes of antibiotics against emerging infectious agents, as well as those agents that may potentially be used as bio-weapons. Completion of these studies will result in a new class of inhibitors with potential antimicrobial activity against Bacillus anthracis, which causes anthrax. The methodologies developed by work supported by this grant will also be potentially applicable to the development of antibiotics against other biomedically important pathogenic bacteria.
