acid-phosphatase and 4-nitroimidazole

acid-phosphatase has been researched along with 4-nitroimidazole* in 1 studies

Other Studies

1 other study(ies) available for acid-phosphatase and 4-nitroimidazole

Article	Year
High throughput screening identified a substituted imidazole as a novel RANK pathway-selective osteoclastogenesis inhibitor. Assay and drug development technologies, 2006, Volume: 4, Issue:4 Receptor activator of nuclear factor-kappaB (NF-kappaB) (RANK) plays a key role in the differentiation, activation, and survival of osteoclasts. Upon activation of RANK with RANK ligand (RANKL), osteoclast precursor cells differentiate into tartrate-resistant acid phosphatase (TRAP)-positive, multinucleated osteoclasts. To identify compounds that block osteoclastogenesis, a cell-based assay was developed using RAW264.7 cells stably transfected with a TRAP promoter-dependent reporter gene as a surrogate readout for differentiation. Described herein is the strategy for high throughput screening and subsequent secondary biological assays for hit triage, which resulted in the identification of compound 1, a 4-nitroimidazole derivative, that specifically inhibited RANKL-induced TRAP gene and protein expression. Compound 1 did not affect the tumor necrosis factor-alpha- or lipopolysaccharide-induced TRAP-luciferase response, suggesting selective inhibition of the RANKL-induced pathway. Reverse transcription polymerase chain reaction analysis confirmed the inhibition of expression of osteoclast marker genes, such as TRAP, cathepsin K, and carbonic anhydrase type II. Compound 1 did not inhibit the RANKL-induced activation of a NF-kappaB reporter gene, or p38 kinase activity, suggesting a mechanism of action downstream of NF-kappaB. Together, these results suggest that we have identified a RANK pathway-specific inhibitor able to block the RANKL-induced osteoclast differentiation process. The hit identification strategy described here can be applied to other cell-based assays using an indirect surrogate readout to improve success rates. Topics: Acid Phosphatase; Animals; Cell Line; Cell Line, Tumor; Cell Survival; Dimethyl Sulfoxide; Dose-Response Relationship, Drug; Enzyme Inhibitors; Gene Expression; Humans; Imidazoles; Isoenzymes; Lipopolysaccharides; Luciferases; Nitroimidazoles; Osteoclasts; Osteoprotegerin; Pyridines; RANK Ligand; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Signal Transduction; Tartrate-Resistant Acid Phosphatase; Tumor Necrosis Factor-alpha	2006

Article

Year

High throughput screening identified a substituted imidazole as a novel RANK pathway-selective osteoclastogenesis inhibitor.

Assay and drug development technologies, 2006, Volume: 4, Issue:4

Receptor activator of nuclear factor-kappaB (NF-kappaB) (RANK) plays a key role in the differentiation, activation, and survival of osteoclasts. Upon activation of RANK with RANK ligand (RANKL), osteoclast precursor cells differentiate into tartrate-resistant acid phosphatase (TRAP)-positive, multinucleated osteoclasts. To identify compounds that block osteoclastogenesis, a cell-based assay was developed using RAW264.7 cells stably transfected with a TRAP promoter-dependent reporter gene as a surrogate readout for differentiation. Described herein is the strategy for high throughput screening and subsequent secondary biological assays for hit triage, which resulted in the identification of compound 1, a 4-nitroimidazole derivative, that specifically inhibited RANKL-induced TRAP gene and protein expression. Compound 1 did not affect the tumor necrosis factor-alpha- or lipopolysaccharide-induced TRAP-luciferase response, suggesting selective inhibition of the RANKL-induced pathway. Reverse transcription polymerase chain reaction analysis confirmed the inhibition of expression of osteoclast marker genes, such as TRAP, cathepsin K, and carbonic anhydrase type II. Compound 1 did not inhibit the RANKL-induced activation of a NF-kappaB reporter gene, or p38 kinase activity, suggesting a mechanism of action downstream of NF-kappaB. Together, these results suggest that we have identified a RANK pathway-specific inhibitor able to block the RANKL-induced osteoclast differentiation process. The hit identification strategy described here can be applied to other cell-based assays using an indirect surrogate readout to improve success rates.

Topics: Acid Phosphatase; Animals; Cell Line; Cell Line, Tumor; Cell Survival; Dimethyl Sulfoxide; Dose-Response Relationship, Drug; Enzyme Inhibitors; Gene Expression; Humans; Imidazoles; Isoenzymes; Lipopolysaccharides; Luciferases; Nitroimidazoles; Osteoclasts; Osteoprotegerin; Pyridines; RANK Ligand; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Signal Transduction; Tartrate-Resistant Acid Phosphatase; Tumor Necrosis Factor-alpha

2006