A Strategy for Discovery of Novel Broad-Spectrum Antibacterials Using a High-Throughput Streptococcus pneumoniae Transcription/Translation Screen

Pratt, Steve D.; David, Caroline A.; Black-Schaefer, Candace; Dandliker, Peter J.; Xuei, Xiaoling; Warrior, Usha; Burns, David J.; Zhong, Ping; Cao, Zhongcheng; Saiki, Anne Y.; Lerner, Claude G.; Chovan, Linda E.; Soni, Niru B.; Nilius, Angela M.; Wagenaar, Frank L.; Merta, Philip J.; Traphagen, Linda; Beutel, Bruce A.

doi:10.1177/1087057103260876

Cited by 30 publications

(33 citation statements)

References 15 publications

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“…Structures of ligands for other ribosomal RNA targets (9 -16) and RNase P (17). 9) Erythromycin, a natural macrolide; 10) telithromycin (Ketek ® , Aventis); 11) linezolid (Zyvox ® , Pharmacia/Pfizer); 12) 6,11-bridged macrolides by Enanta Pharmaceuticals [239,242]; 13) synthetic thiostrepton fragment by Ribotargets [40]; 14) synthetic thiostrepton mimetic by Isis Pharmaceuticals [28]; 15) fluoroquinolone translation inhibitor by Abbott Laboratories [45]; 16) 14-membered macrocyclic translation inhibitor by Isis Pharmaceuticals [47]; 17) aromatic bis-guanylhydrazone RNase P inhibitor by Message Pharmaceuticals [258,259]. For discussion, see text.…”

Section: Other Ribosomal Targets and Ligandsmentioning

confidence: 99%

RNA as a target for small-molecule therapeutics

Hermann¹,

Tor²

2005

Expert Opinion on Therapeutic Patents

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Natural antibiotics that specifically target RNA components of the bacterial ribosome set precedence for RNA-directed small-molecule drugs. Structured domains of functional RNAs in bacteria and viruses have thus attracted attention as targets for the discovery of novel anti-infectives. Whereas a growing number of RNA-directed ligands have been reported in the literature, creating synthetic molecules that combine high affinity with selectivity for a specific RNA target remains a key challenge. Ongoing discovery efforts towards this goal, combined with the increasing knowledge of the factors that govern small-molecule recognition of RNA folds, will prepare the ground for the development of novel RNA-targeted therapeutics.

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Section: Other Ribosomal Targets and Ligandsmentioning

confidence: 99%

RNA as a target for small-molecule therapeutics

Hermann¹,

Tor²

2005

Expert Opinion on Therapeutic Patents

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“…Of the many possible applications for this technology, perhaps one of the most important is that it facilitates an ideal approach to the discovery of novel antibacterial agents. From a practical standpoint, researchers have had to choose between two extremes: cellular screening and molecular target screening, although pathway screens with cell extracts have also been pursued (12,24,35). Screening directly for inhibitors of bacterial proliferation results in compounds with antibacterial activity, but without a readily discovered molecular target to facilitate structure and/or mechanism-directed medicinal chemistry for lead optimization.…”

Section: Figmentioning

confidence: 99%

Novel Approach to Mapping of Resistance Mutations in Whole Genomes by Using Restriction Enzyme Modulation of Transformation Efficiency

Lerner

Kakavas

Wagner

et al. 2005

Antimicrob Agents Chemother

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Restriction enzyme modulation of transformation efficiencies (REMOTE) is a method that makes use of genome restriction maps and experimentally observed differences in transformation efficiencies of genomic DNA restriction digests to discover the location of mutations in genomes. The frequency with which digested genomic DNA from a resistant strain transforms a susceptible strain to resistance is primarily determined by the size of the fragment containing the resistance mutation and the distance of the mutation to the end of the fragment. The positions of restriction enzyme cleavage sites immediately flanking the resistance mutation define these parameters. The mapping procedure involves a process of elimination in which digests that transform with high frequency indicate that the restriction enzyme cleavage sites are relatively far away from the mutation, while digests that transform with low frequency indicate that the sites are close to the mutation. The transformation data are compared computationally to the genome restriction map to identify the regions that best fit the data. Transformations with PCR amplicons encompassing candidate regions identify the resistance locus and enable identification of the mutation. REMOTE was developed using Haemophilus influenzae strains with mutations in gyrA, gyrB, and rpsE that confer resistance to ciprofloxacin, novobiocin, and spectinomycin, respectively. We applied REMOTE to identify mutations that confer resistance to two novel antibacterial compounds. The resistance mutations were found in genes that can decrease the intracellular concentration of compounds: acrB, which encodes a subunit of the AcrAB-TolC efflux pump; and fadL, which encodes a long-chain fatty acid transporter.

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“…Recently, attempts have been made to screen chemical-compound libraries by using cell extracts containing native transcription and translation systems from Escherichia coli (37), Streptococcus pneumoniae (9,37), and Staphylococcus aureus (28). This approach has had only limited success.…”

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confidence: 99%

Discovery and Analysis of 4 H -Pyridopyrimidines, a Class of Selective Bacterial Protein Synthesis Inhibitors

Ribble

Hill

Ochsner

et al. 2010

Antimicrob Agents Chemother

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Bacterial protein synthesis is the target for numerous natural and synthetic antibacterial agents. We have developed a poly(U) mRNA-directed aminoacylation/translation protein synthesis system composed of phenyltRNA synthetases, ribosomes, and ribosomal factors from Escherichia coli. This system, utilizing purified components, has been used for high-throughput screening of a small-molecule chemical library. We have identified a series of compounds that inhibit protein synthesis with 50% inhibitory concentrations (IC 50 s) ranging from 3 to 14 M. This series of compounds all contained the same central scaffold composed of tetrahydropyrido[4,3-d]pyrimidin-4-ol (e.g., 4H-pyridopyrimidine). All analogs contained an ortho pyridine ring attached to the central scaffold in the 2 position and either a five-or a six-member ring tethered to the 6-methylene nitrogen atom of the central scaffold. These compounds inhibited the growth of E. coli, Staphylococcus aureus, Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis, with MICs ranging from 0.25 to 32 g/ml. Macromolecular synthesis (MMS) assays with E. coli and S. aureus confirmed that antibacterial activity resulted from specific inhibition of protein synthesis. Assays were developed for the steps performed by each component of the system in order to ascertain the target of the compounds, and the ribosome was found to be the site of inhibition.

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A Strategy for Discovery of Novel Broad-Spectrum Antibacterials Using a High-Throughput Streptococcus pneumoniae Transcription/Translation Screen

Cited by 30 publications

References 15 publications

RNA as a target for small-molecule therapeutics

RNA as a target for small-molecule therapeutics

Novel Approach to Mapping of Resistance Mutations in Whole Genomes by Using Restriction Enzyme Modulation of Transformation Efficiency

Discovery and Analysis of 4 H -Pyridopyrimidines, a Class of Selective Bacterial Protein Synthesis Inhibitors

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