Description:
Researchers at MUSC have developed a high-throughput assay that uses fluorescence polarization (FP) to distinguish the fluorescent penicillin, Bocillin-FL, in free or PBP-bound form. The assay can be used to identify new compounds that inhibit penicillin binding proteins (PBPs), which are proven targets for β-lactam antibiotics. This assay was used to screen a 50,000 compound library for potential inhibitors of N. gonorrhoeae PBP 2. As a result, 32 compounds were identified that exhibited >50% inhibition of Bocillin-FL binding to PBP 2. These included a cephalosporin that provided validation of the assay. This assay paves the way for more comprehensive high-throughput screening of compound libraries for non-β-lactam antimicrobial compounds.
A high-throughput screen of 50,000 compounds was performed to identify PBP binding and inhibiting compounds.
Overview:
Over the past several years, the prevalence of gonorrhea infection within the US has increased, with the CDC reporting over 321,000 infections in 2011. In women, infections from gonorrhea are a major cause of pelvic inflammatory disease, which can have serious complications including infertility, ectopic pregnancy and chronic pelvic pain. Additionally, gonorrhea has been shown to facilitate HIV infection. The increasing prevalence of Neisseria gonorrhoeae strains exhibiting decreased susceptibility to third-generation cephalosporins and the recent isolation of two distinct strains with high-level resistance to cefixime or ceftriaxone heralds the possible demise of β-lactam antibiotics as effective treatments for gonorrhea. Due to steadily increasing resistance to the third generation of cephalosporins, treatment of gonorrhea is on its last line of defense, with the only recognized, suitable treatment a combination therapy.
Advantages: High-throughput assay for new antimicrobials
Key Words: Neisseria gonorrhoeae, gonorrhea, bacteria, antibiotics, antimicrobials, drug resistance, assay, penicillin binding protein, PBP
Publications: Fedarovich, Alena, et al. "High-throughput screening for novel inhibitors of Neisseria gonorrhoeae penicillin-binding protein 2." PLoS One 7.9 (2012): e44918.
Inventors: C. Davis and A. Fedarovich
Patent Status: PCT Application filed 08/01/2013, PCT/US2013/053127
MUSC-FRD Technology ID: P1254