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G. Hoppe, Q. Ebrahem, J. E. Sears; A Novel Class of Small Molecules for Treatment of Retinopathy of Prematurity. Invest. Ophthalmol. Vis. Sci. 2009;50(13):3126.
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© ARVO (1962-2015); The Authors (2016-present)
To expand the search for a pharmaceutical treatment of retinopathy of prematurity based on our recent observation of preventing oxygen induced vascular obliteration by activation of hypoxia inducible factor 1 (HIF-1). In this study we have conducted high throughput large scale screening of the Chembridge small molecule library for compounds capable of upregulating HIF-1.
A Chemical DiverSet library of 34,000 chemical compounds consisted of handcrafted drug-like organic molecules with molecular weight in a range of 250 - 550. Primary screening was performed using NIH3T3 cell line stably expressing reporter gene of firefly luciferase under a HIF-1-inducible promoter. Hierarchical multi-step validation of the reporter gene activators was performed by assessing their effect on endogenous HIF-1 levels and erythropoietin (Epo) secretion in cell cultures.
We have identified and selected 23 potential HIF inducers that demonstrate ability to stimulate HIF-dependent transcription at list 3-fold. Six compounds increased Epo protein levels while only four compounds substantially induced HIF-1 levels in cultured cells. One compound lead to HIF-1 upregulation that did not lead to Epo expression and three other compounds caused Epo production, but only minimal induction of HIF-1. As a result of secondary screening we have selected 3 compounds that induce robust increase of intracellular HIF-1 and secreted Epo by cultures cells.
A striking number of compounds capable of HIF-1 upregulation were from a similar family of small molecules. Furthermore, newly identified HIF activators were potent at relatively low micromolar concentrations. Finally, their differential effects on HIF and Epo induction provide us with a molecular tool to dissect the pathways involved in the protection of the neurovascular unit.
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