title: Activity-guided screening of bioactive natural compounds implementing a new glucocorticoid-receptor-translocation assay and detection of new anti-inflammatory steroids from bacteria.
abstract: Using an in vitro cell-based assay in a flow-design, we have applied activity-guided screening to search for new bioactive compounds isolated from microorganisms. A first assay employs the stable expression of nuclear factor kappa B (NF-κB) while a second assay utilizes the glucocorticoid receptor (GR) coupled to green fluorescent protein. A specialized assay was implemented for both the translocation of NF-κB and to inhibit the translocation of cytokine-mediated NF-κB. In addition, we developed in a wide palette of cell lines used for a highly specialized GR-translocation assay to detect anti-inflammatory effects. This approach demonstrates the straight-forward combination of cell-based assays arranged with an automated fluorescence microscope. This allows for the direct sorting of extracts which are acting in a pharmaceutically interesting way. Initial results using this technique have led to the detection of new anti-inflammatory steroids from bacterial crude extracts.
authors.affiliation: Department of Pharmaceutical Biotechnology, Saarland University, Saarbrücken, Germany. kaufm@uni-trier.de
authors.forename: Katrin
authors.initials: K
authors.lastname: Kaufmann
authors.forename: Luke
authors.initials: L
authors.lastname: Simmons
authors.forename: Jennifer
authors.initials: J
authors.lastname: Herrmann
authors.forename: Gertrud
authors.initials: G
authors.lastname: Schwär
authors.forename: Nora
authors.initials: N
authors.lastname: Luniak
authors.forename: Rolf
authors.initials: R
authors.lastname: Müller
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000378:metabolism / Q000494:pharmacology*; D001681:Biological Assay / Q000379:methods*; D001688:Biological Products / Q000378:metabolism / Q000494:pharmacology*; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006914:Hydroxysteroids / Q000378:metabolism / Q000494:pharmacology*; D051379:Mice; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D000893:Anti-Inflammatory Agents; D001688:Biological Products; D006914:Hydroxysteroids; D016328:NF-kappa B; D017382:Reactive Oxygen Species; D011965:Receptors, Glucocorticoid; D049452:Green Fluorescent Proteins