title: Neuroprotection by small molecule activators of the nerve growth factor receptor.
abstract: There is a great deal of interest in neurotrophin therapy to prevent neuronal degeneration. However, the blood-brain barrier presents a major hurdle in the use of peptide therapeutics. The goal of this study was to identify small molecule, cell-permeable nerve growth factor (NGF) activators. Combinatorial libraries of asterriquinones (>300) and mono-indolyl-quinones (>60) were screened using a 96-well enzyme-linked immunosorbent assay that detects phosphorylated TrkA, the NGF receptor. The libraries were also screened for dose-dependent cytotoxicity. From these screens, we generated quantitative structure-activity relationship models for activity and toxicity, and then we selected two compounds, 2-(6-chloro-1H-indol-3-yl)-5-(2-cyclopropyl-1H-indol-3-yl)-3,6-dihydroxy-[1,4]benzoquinone (1H5) and 2,5-dimethoxy-3-(7-fluoro-1H-indol-3-yl)-[1,4]-benzoquinone (5E5), for further study based on high activity and low toxicity. Compound 1H5 (30 microM) is an asterriquinone that is a moderate TrkA activator (50% the activity of 100 ng/ml NGF), and it shows little toxicity at concentrations up to 100 microM. 1H5 can protect differentiated PC12 neurons from apoptotic cell death induced by NGF withdrawal. Compound 5E5 (30 microM) is a mono-indolyl-quinone that is a very strong activator of TrkA (>200% the activity of 100 ng/ml NGF), and it is nontoxic at concentrations up to 10 microM. Activation of TrkA can be detected at 1 microM 5E5, and 3 to 10 microM 5E5 activates TrkA and extracellular signal-regulated kinase as strongly as a maximal dose of NGF (100 ng/ml). A combination of a low dose of 5E5 (1 microM) with a submaximal dose of NGF (10 ng/ml) promotes neuronal differentiation of PC12 cells. These compounds represent a new class of TrkA activators that could have potential utility in the treatment of neurodegenerative diseases.
authors.affiliation: Veterans Medical Research Foundation and Veterans Affairs San Diego Healthcare System, San Diego, California, USA.
authors.forename: Bo
authors.initials: B
authors.lastname: Lin
authors.forename: Michael C
authors.initials: MC
authors.lastname: Pirrung
authors.forename: Liu
authors.initials: L
authors.lastname: Deng
authors.forename: Zhitao
authors.initials: Z
authors.lastname: Li
authors.forename: Yufa
authors.initials: Y
authors.lastname: Liu
authors.forename: Nicholas J G
authors.initials: NJ
authors.lastname: Webster
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D016466:CHO Cells; D002454:Cell Differentiation / Q000187:drug effects; D020650:Combinatorial Chemistry Techniques; D006224:Cricetinae; D003412:Cricetulus; D020935:MAP Kinase Signaling System; D018696:Neuroprotective Agents / Q000494:pharmacology*; D016716:PC12 Cells; D021281:Quantitative Structure-Activity Relationship; D051381:Rats; D020917:Receptor, trkA / Q000187:drug effects; D017475:Receptors, Nerve Growth Factor / Q000187:drug effects*
chemical_list: D018696:Neuroprotective Agents; D017475:Receptors, Nerve Growth Factor; D020917:Receptor, trkA