title: Identification and validation of small molecule modulators of the NusB-NusE interaction.
abstract: Formation of highly possessive antitermination complexes is crucial for the efficient transcription of stable RNA in all bacteria. A key step in the formation of these complexes is the protein-protein interaction (PPI) between N-utilisation substances (Nus) B and E and thus this PPI offers a novel target for a new antibiotic class. A pharmacophore developed via a secondary structure epitope approach was utilised to perform an in silico screen of the mini-Maybridge library (56,000 compounds) which identified 25 hits of which five compounds were synthetically tractable leads. Here we report the synthesis of these five leads and their biological evaluation as potential inhibitors of the NusB-NusE PPI. Two chemically diverse scaffolds were identified to be low micro molar potent PPI inhibitors, with compound (4,6-bis(2',4',3.4 tetramethoxyphenyl))pyrimidine-2-sulphonamido-N-4-acetamide 1 and N,N'-[1,4-butanediylbis(oxy-4,1-phenylene)]bis(N-ethyl)urea 3 exhibiting IC50values of 6.1μM and 19.8μM, respectively. These inhibitors were also shown to be moderate inhibitors of Gram-positive Bacillus subtilis and Gram-negative Escherichia coli growth.
authors.affiliation: Chemistry, School of Environmental and Life Sciences, University of Newcastle, University Drive, Callaghan, NSW 2308, Australia.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Cossar
authors.affiliation: Biology, Centre for Chemical Biology, School of Environmental and Life Sciences, University of Newcastle, University Drive, Callaghan, NSW 2308, Australia.
authors.forename: Cong
authors.initials: C
authors.lastname: Ma
authors.affiliation: Chemistry, School of Environmental and Life Sciences, University of Newcastle, University Drive, Callaghan, NSW 2308, Australia.
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Gordon
authors.affiliation: Chemistry, School of Environmental and Life Sciences, University of Newcastle, University Drive, Callaghan, NSW 2308, Australia.
authors.forename: Joseph I
authors.initials: JI
authors.lastname: Ambrus
authors.affiliation: Biology, Centre for Chemical Biology, School of Environmental and Life Sciences, University of Newcastle, University Drive, Callaghan, NSW 2308, Australia.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Lewis
authors.affiliation: Chemistry, School of Environmental and Life Sciences, University of Newcastle, University Drive, Callaghan, NSW 2308, Australia. Electronic address: Adam.McCluskey@newcastle.edu.au.
authors.forename: Adam
authors.initials: A
authors.lastname: McCluskey
mesh_terms: D000900:Anti-Bacterial Agents / Q000138:chemical synthesis / Q000494:pharmacology*; D001412:Bacillus subtilis / Q000187:drug effects; D004926:Escherichia coli / Q000187:drug effects; D029968:Escherichia coli Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry; D006860:Hydrogen Bonding; D009842:Oligopeptides / Q000737:chemistry / Q000494:pharmacology; D010647:Phenyl Ethers / Q000138:chemical synthesis / Q000494:pharmacology; D011485:Protein Binding; D011743:Pyrimidines / Q000138:chemical synthesis / Q000494:pharmacology; D012269:Ribosomal Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry; D013449:Sulfonamides / Q000138:chemical synthesis / Q000494:pharmacology; D014157:Transcription Factors / Q000037:antagonists & inhibitors* / Q000737:chemistry
chemical_list: D000900:Anti-Bacterial Agents; D029968:Escherichia coli Proteins; C074952:NusB protein, E coli; D009842:Oligopeptides; D010647:Phenyl Ethers; D011743:Pyrimidines; D012269:Ribosomal Proteins; D013449:Sulfonamides; D014157:Transcription Factors; C000622945:YDHRLLDQS-NH2; C029537:ribosomal protein S10
keywords: Antibiotic; In silico screening; NusB-NusE; Pharmacophore; Protein-protein interaction