title: Novel inhibitors of DNA gyrase: 3D structure based biased needle screening, hit validation by biophysical methods, and 3D guided optimization. A promising alternative to random screening.
abstract: Random screening provided no suitable lead structures in a search for novel inhibitors of the bacterial enzyme DNA gyrase. Therefore, an alternative approach had to be developed. Relying on the detailed 3D structural information of the targeted ATP binding site, our approach combines as key techniques (1) an in silico screening for potential low molecular weight inhibitors, (2) a biased high throughput DNA gyrase screen, (3) validation of the screening hits by biophysical methods, and (4) a 3D guided optimization process. When the in silico screening was performed, the initial data set containing 350 000 compounds could be reduced to 3000 molecules. Testing these 3000 selected compounds in the DNA gyrase assay provided 150 hits clustered in 14 classes. Seven classes could be validated as true, novel DNA gyrase inhibitors that act by binding to the ATP binding site located on subunit B: phenols, 2-amino-triazines, 4-amino-pyrimidines, 2-amino-pyrimidines, pyrrolopyrimidines, indazoles, and 2-hydroxymethyl-indoles. The 3D guided optimization provided highly potent DNA gyrase inhibitors, e. g., the 3,4-disubstituted indazole 23 being a 10 times more potent DNA gyrase inhibitor than novobiocin (3).
authors.affiliation: Preclinical Research, Pharmaceuticals Division, F. Hoffmann-La Roche Ltd., CH-4070 Basel, Switzerland.
authors.forename: H J
authors.initials: HJ
authors.lastname: Boehm
authors.forename: M
authors.initials: M
authors.lastname: Boehringer
authors.forename: D
authors.initials: D
authors.lastname: Bur
authors.forename: H
authors.initials: H
authors.lastname: Gmuender
authors.forename: W
authors.initials: W
authors.lastname: Huber
authors.forename: W
authors.initials: W
authors.lastname: Klaus
authors.forename: D
authors.initials: D
authors.lastname: Kostrewa
authors.forename: H
authors.initials: H
authors.lastname: Kuehne
authors.forename: T
authors.initials: T
authors.lastname: Luebbers
authors.forename: N
authors.initials: N
authors.lastname: Meunier-Keller
authors.forename: F
authors.initials: F
authors.lastname: Mueller
mesh_terms: D000890:Anti-Infective Agents / Q000138:chemical synthesis / Q000737:chemistry*; D003374:Coumarins / Q000138:chemical synthesis / Q000737:chemistry; D018360:Crystallography, X-Ray; D004250:DNA Topoisomerases, Type II / Q000737:chemistry*; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry*; D007191:Indazoles / Q000138:chemical synthesis / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D015394:Molecular Structure; D009675:Novobiocin / Q000737:chemistry; D011485:Protein Binding; D013329:Structure-Activity Relationship; D020349:Surface Plasmon Resonance; D059005:Topoisomerase II Inhibitors; D014461:Ultracentrifugation
chemical_list: C411457:7-(4-(4-tert-butylbenzyloxy)-1H-indazol-3-ylmethylsulfanyl)-4-methylcoumarin; D000890:Anti-Infective Agents; D003374:Coumarins; D004791:Enzyme Inhibitors; D007191:Indazoles; D059005:Topoisomerase II Inhibitors; D009675:Novobiocin; D004250:DNA Topoisomerases, Type II