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Breandan N Kennedy, Yolanda Alvarez, Claire Kilty, Orla Galvin, Clare Butler, Stephanie Merrigan, Temitope Sasore, Carmel McVicar, Alan W Stitt, Alison Reynolds; Phenotype-Based Discovery of Cysteinyl Leukotriene Receptor Antagonists as Novel Inhibitors of Developmental and Pathological Ocular Angiogenesis.. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):5848.
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© ARVO (1962-2015); The Authors (2016-present)
Developmental angiogenesis generates the ocular vasculature that nourishes the eye. In proliferative diabetic retinopathy and neovascular age-related macular degeneration, neovascularisation is part of the pathological processes leading to blindness. Our goal is to discover novel drugs that inhibit developmental and pathological angiogenesis in the eye, to better understand the molecular processes regulating ocular angiogenesis and to improve treatment options for proliferative ocular diseases.
Phenotype-based screens were performed using the Chembridge Diverset library. Drugs were screened for inhibition of hyaloid vessel (HV) angiogenesis in Tg(fli1:EGFP) zebrafish. Hits were screened for anti-angiogenic activity in human endothelial cells (HMEC-1), in rat aortic ring explants and in the mouse oxygen-induced retinopathy (OIR) model. Safety pharmacology was assessed by visual behaviour assays, light microscopy analysis of retinal sections and cytotoxicity profiling of human ARPE-19 and HMEC-1. Cysteinyl leukotriene receptor (cysLTR1/2) expression was analysed by Western blot and RT-PCR. CysLTR1/2 antagonism was quantified by lung strip contraction and cell-based flourescence assays.
A hit molecule, 2-[(E)-2-(Quinolin-2-yl)vinyl]phenol, designated Quininib robustly inhibits developmental angiogenesis at 5-10 uM in zebrafish and was subsequently shown to significantly inhibit angiogenic tubule formation in HMEC-1 cells, angiogeneic sprouting in aortic ring explants and retinal neovascularisation in OIR mice. 5-10 uM quininib is well tolerated in zebrafish visual behaviour assays and 200 uM intravitreal quininib has no adverse drug reaction on murine retinal morphology. Profiling screens of ~200 angiogenic and inflammatory targets revealed quininib not to target VEGF receptors, but to antagonise cysLTR1 with an IC50 of ~ 4 uM. CysLTR1/2 are expressed in the retina during neovascularisation.
Phenotype-based drugs screens identified Quininib, a novel anti-angiogenic small molecule. Quininib, a cysteinyl leukotriene receptor antagonist, inhibits developmental and pathological angiognesis. Quinininb reveals a novel role for cysLTR1 in ocular developmental angiogenesis and offers potential to treat patients non-responsive to current anti-VEGF treatments or to enhance efficacy in combination with anti-VEGF biologicals.
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