title: Development of an AlphaScreen-based high-throughput screening assay for inhibitors of human vascular endothelial growth factor receptor-3.
abstract: Vascular endothelial growth factor receptor 3 (VEGFR-3, also known as Flt4), the receptor for vascular endothelial growth factors (VEGFs) C and D, is expressed on lymphatic endothelium and plays an important role in lymphangiogenesis. Recent studies indicate that VEGFR-3 may be involved in tumor angiogenesis and growth, and so VEGFR-3 is an attractive target for researchers seeking to prevent tumor growth and metastasis. Hence we expressed and purified the CD (catalytic domain) of VEGFR-3 (VEGFR-3-CD) in Escherichia coli expression system and established a new simple and effective HTS (high-throughput screening) assay based on AlphaScreen strategy for VEGFR-3 inhibitors. Then we identified a novel cobalt complex that inhibited the phosphorylation of VEGFR-3 in a dose-dependent manner with an IC(50) value of 338 nM. This compound could be a useful new angiogenesis inhibitor, which would be investigated further for its therapeutic potentials.
authors.affiliation: Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu Province, PR China.
authors.forename: Ke
authors.initials: K
authors.lastname: Wang
authors.forename: Runlin
authors.initials: R
authors.lastname: Yang
authors.forename: Juan
authors.initials: J
authors.lastname: Xu
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.forename: Lan
authors.initials: L
authors.lastname: Zhu
authors.forename: Jianguo
authors.initials: J
authors.lastname: Lin
authors.forename: Biao
authors.initials: B
authors.lastname: Huang
mesh_terms: D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007211:Indoles / Q000032:analysis* / Q000494:pharmacology*; D011758:Pyrroles / Q000032:analysis* / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013329:Structure-Activity Relationship; D000077210:Sunitinib; D040321:Vascular Endothelial Growth Factor Receptor-3 / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology
chemical_list: D007211:Indoles; D011758:Pyrroles; D040321:Vascular Endothelial Growth Factor Receptor-3; D000077210:Sunitinib