Folate biosynthesis is an essential bacterial pathway that is absent in higher animals, and it has been a target of antibacterial agents for over 70 years. Sulfa drugs inhibit the enzyme dihydropteroate synthase (DHPS) in the pathway by acting as non-productive substrate analogs of p-aminobenzoic acid (pABA). However, the flexible pABA binding site is structurally susceptible to resistance mutations, and the sulfa drugs are rapidly becoming therapeutically ineffective. In contrast, the binding site of the second DHPS substrate, pterin-pyrophosphate, is buried in a conserved pocket that is less likely to tolerate mutations. We propose to generate new classes of potent DHPS inhibitors that specifically engage this pocket. These inhibitors can potentially be developed into novel therapeutic agents that still target folate synthesis but which avoid the problems of resistance. To generate effective inhibitors of any enzyme, it is crucial to understand the structure and mechanism of its active site. This information is largely absent for DHPS, and understanding how DHPS performs catalysis at the molecular level will be a central goal of the proposal. Another goal will be to understand the molecular and functional basis of sulfa drug resistance to avoid this problem in our future drug designs. This comprehensive approach will require the joint expertise of the two principal investigators in structural biology, medicinal chemistry, biochemistry, computational biology and microbiology. The project will also incorporate the relatively new technique of fragment based drug discovery to identify novel small inhibitory chemical scaffolds that can readily be elaborated by chemical synthesis. Exciting new data described in the application provide strong support that the project can realize its ultimate goal which is to rapidly develop new antimicrobials that minimize the emergence of drug resistance. Although we aim to develop broad-spectrum anti-infective agents, our microbiological screening will focus on S. aureus and P. jirovecii that have emerged as particularly problematical infectious disease agents in the U.S. in recent years.

Public Health Relevance

This proposal is aimed at structurally understanding and exploiting the enzyme target of the sulfonamide antibacterial agents, dihydropteroate synthase;to study enzyme mechanism, drug resistance and inhibitor design. It is expected that these studies will lead to new antimicrobial agents and a new understanding of target based sulfonamide resistance.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI070721-06
Application #
8245291
Study Section
Special Emphasis Panel (ZRG1-DDR-T (09))
Program Officer
Franceschi, Francois J
Project Start
2006-06-01
Project End
2016-06-30
Budget Start
2012-07-15
Budget End
2013-06-30
Support Year
6
Fiscal Year
2012
Total Cost
$387,209
Indirect Cost
$156,651
Name
St. Jude Children's Research Hospital
Department
Type
DUNS #
067717892
City
Memphis
State
TN
Country
United States
Zip Code
38105
Yun, Mi-Kyung; Hoagland, Daniel; Kumar, Gyanendra et al. (2014) The identification, analysis and structure-based development of novel inhibitors of 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase. Bioorg Med Chem 22:2157-65
Zhao, Ying; Hammoudeh, Dalia; Yun, Mi-Kyung et al. (2012) Structure-based design of novel pyrimido[4,5-c]pyridazine derivatives as dihydropteroate synthase inhibitors with increased affinity. ChemMedChem 7:861-70
Qi, Jianjun; Virga, Kristopher G; Das, Sourav et al. (2011) Synthesis of bi-substrate state mimics of dihydropteroate synthase as potential inhibitors and molecular probes. Bioorg Med Chem 19:1298-305
Zhao, Ying; Hammoudeh, Dalia; Lin, Wenwei et al. (2011) Development of a pterin-based fluorescent probe for screening dihydropteroate synthase. Bioconjug Chem 22:2110-7
Hevener, Kirk E; Yun, Mi-Kyung; Qi, Jianjun et al. (2010) Structural studies of pterin-based inhibitors of dihydropteroate synthase. J Med Chem 53:166-77
Hevener, Kirk E; Zhao, Wei; Ball, David M et al. (2009) Validation of Molecular Docking Programs for Virtual Screening against Dihydropteroate Synthase. J Chem Inf Model :
Hevener, Kirk E; Zhao, Wei; Ball, David M et al. (2009) Validation of molecular docking programs for virtual screening against dihydropteroate synthase. J Chem Inf Model 49:444-60
Yun, Mi-Kyung; Nourse, Amanda; White, Stephen W et al. (2007) Crystal structure and allosteric regulation of the cytoplasmic Escherichia coli L-asparaginase I. J Mol Biol 369:794-811