title: Microplate-based screening for small molecule inhibitors of neuropilin-2/vascular endothelial growth factor-C interactions.
abstract: Vascular endothelial growth factor-C (VEGF-C) is a secreted growth factor essential for lymphangiogenesis. VEGF-C functions in both physiological and pathological lymphangiogenesis, particularly in tumor metastasis, making it an attractive therapeutic target. Members of two families of cell surface receptors transduce VEGF-C signals: neuropilin-2 (Nrp2) and VEGF-receptor (VEGFR)-2/3. Nrp2 is a promising target for inhibition because it is highly expressed in lymphatic vessels. Here we describe a microplate-based assay for discovery of VEGF-C/Nrp2 inhibitors. We optimize this assay for use in screening an inhibitor library and identify three novel Nrp2/VEGF-C binding inhibitors from the National Institutes of Health (NIH) Clinical Collection small molecule library.
authors.affiliation: Center for Structural Biology, Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, KY 40536, USA.
authors.forename: Matthew W
authors.initials: MW
authors.lastname: Parker
authors.affiliation: Center for Structural Biology, Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, KY 40536, USA. Electronic address: craig.vanderkooi@uky.edu.
authors.forename: Craig W
authors.initials: CW
authors.lastname: Vander Kooi
mesh_terms: D003609:Dactinomycin / Q000737:chemistry / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D055808:Drug Discovery; D057166:High-Throughput Screening Assays; D006801:Humans; D007211:Indoles; D009316:National Institutes of Health (U.S.); D039943:Neuropilin-2 / Q000737:chemistry / Q000378:metabolism*; D010617:Phenanthridines / Q000737:chemistry / Q000494:pharmacology; D048448:Phenylcarbamates; D011485:Protein Binding / Q000187:drug effects; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013449:Sulfonamides; D014105:Tosyl Compounds / Q000737:chemistry / Q000494:pharmacology; D014481:United States; D042582:Vascular Endothelial Growth Factor C / Q000737:chemistry / Q000378:metabolism*
chemical_list: D007211:Indoles; D039943:Neuropilin-2; D010617:Phenanthridines; D048448:Phenylcarbamates; D054852:Small Molecule Libraries; D013449:Sulfonamides; D014105:Tosyl Compounds; C467486:VEGFC protein, human; D042582:Vascular Endothelial Growth Factor C; C581308:neuropilin-2, human; D003609:Dactinomycin; C061532:dihydrexidine; C062735:zafirlukast
keywords: Assay; Inhibitor; Ligand; Neuropilin; Receptor; VEGF