{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Scharf NT"],"funding":["NIAID NIH HHS","NIGMS NIH HHS"],"pagination":["287-297"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC5323270"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["22(3)"],"pubmed_abstract":["Rifampin has been a cornerstone of tuberculosis (TB) treatment since its introduction. The rise of multidrug-resistant and extensively drug-resistant TB makes the development of novel therapeutics effective against these strains an urgent need. Site-specific mutations in the target enzyme of rifampin, RNA polymerase (RNAP) comprises the majority (~97%) of rifamycin-resistant (Rif<sup>R</sup>) strains of Mycobacterium tuberculosis (MTB). To identify novel inhibitors of bacterial RNAP, an in vitro plasmid-based transcription assay that uses malachite green (MG) to detect transcribed RNA containing MG aptamers was developed. This assay was optimized in a 384-well plate format and used to screen 150,000 compounds against an Escherichia coli homolog of the most clinically relevant Rif<sup>R</sup> RNAP (βS531L) containing a mutation (β'V408G) that compensates for the fitness defect of this Rif<sup>R</sup> mutant. Following confirmation and concentration-response studies, 10 compounds were identified with similar in vitro inhibition values across a panel of wild-type and Rif<sup>R</sup> E. coli and MTB RNAPs. Four compounds identified from the screen are active against MTB in culture at concentrations below 200 µM. Initial follow-up has resulted in the elimination of one scaffold due to potential pan-assay interference."],"journal":["SLAS discovery : advancing life sciences R & D"],"pubmed_title":["Novel Chemical Scaffolds for Inhibition of Rifamycin-Resistant RNA Polymerase Discovered from High-Throughput Screening."],"pmcid":["PMC5323270"],"funding_grant_id":["R01 AI110780","T32 GM007767"],"pubmed_authors":["Garcia GA","Molodtsov V","Kontos A","Murakami KS","Scharf NT"],"additional_accession":[]},"is_claimable":false,"name":"Novel Chemical Scaffolds for Inhibition of Rifamycin-Resistant RNA Polymerase Discovered from High-Throughput Screening.","description":"Rifampin has been a cornerstone of tuberculosis (TB) treatment since its introduction. The rise of multidrug-resistant and extensively drug-resistant TB makes the development of novel therapeutics effective against these strains an urgent need. Site-specific mutations in the target enzyme of rifampin, RNA polymerase (RNAP) comprises the majority (~97%) of rifamycin-resistant (Rif<sup>R</sup>) strains of Mycobacterium tuberculosis (MTB). To identify novel inhibitors of bacterial RNAP, an in vitro plasmid-based transcription assay that uses malachite green (MG) to detect transcribed RNA containing MG aptamers was developed. This assay was optimized in a 384-well plate format and used to screen 150,000 compounds against an Escherichia coli homolog of the most clinically relevant Rif<sup>R</sup> RNAP (βS531L) containing a mutation (β'V408G) that compensates for the fitness defect of this Rif<sup>R</sup> mutant. Following confirmation and concentration-response studies, 10 compounds were identified with similar in vitro inhibition values across a panel of wild-type and Rif<sup>R</sup> E. coli and MTB RNAPs. Four compounds identified from the screen are active against MTB in culture at concentrations below 200 µM. Initial follow-up has resulted in the elimination of one scaffold due to potential pan-assay interference.","dates":{"release":"2017-01-01T00:00:00Z","publication":"2017 Mar","modification":"2024-02-15T01:27:13.71Z","creation":"2019-03-26T23:04:08Z"},"accession":"S-EPMC5323270","cross_references":{"pubmed":["28027449"],"doi":["10.1177/2472555216679994"]}}