Abstract
EWS-FLI1 is an oncogenic fusion protein implicated in the development of Ewing's sarcoma family tumors (ESFT). Using our previously reported lead compound 2 (YK-4-279), we designed and synthesized a focused library of analogues. The functional inhibition of the analogues was measured by an EWS-FLI1/NR0B1 reporter luciferase assay and a paired cell screening approach measuring effects on growth inhibition for human cells containing EWS-FLI1 (TC32 and TC71) and control PANC1 cell lines devoid of the oncoprotein. Our data revealed that substitution of electron donating groups at the para-position on the phenyl ring was the most favorable for inhibition of EWS-FLI1 by analogs of 2. Compound 9u (with a dimethylamino substitution) was the most active inhibitor with GI50 = 0.26 ± 0.1 μM. Further, a correlation of growth inhibition (EWS-FLI1 expressing TC32 cells) and the luciferase reporter activity was established (R(2) = 0.84). Finally, we designed and synthesized a biotinylated analogue and determined the binding affinity for recombinant EWS-FLI1 (Kd = 4.8 ± 2.6 μM).
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Aniline Compounds / chemical synthesis
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Aniline Compounds / pharmacology*
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Antineoplastic Agents / chemical synthesis*
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Antineoplastic Agents / pharmacology*
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Bone Neoplasms / drug therapy*
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Bone Neoplasms / metabolism
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Bone Neoplasms / pathology
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Cell Proliferation / drug effects
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Crystallography, X-Ray
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Gene Expression Regulation, Neoplastic / drug effects*
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Humans
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Indoles / chemical synthesis
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Indoles / pharmacology*
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Luciferases / metabolism
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Models, Molecular
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Molecular Structure
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Oncogene Proteins, Fusion / antagonists & inhibitors*
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Oncogene Proteins, Fusion / metabolism
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Proto-Oncogene Protein c-fli-1 / antagonists & inhibitors*
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Proto-Oncogene Protein c-fli-1 / metabolism
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RNA-Binding Protein EWS / antagonists & inhibitors*
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RNA-Binding Protein EWS / metabolism
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Sarcoma, Ewing / drug therapy*
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Sarcoma, Ewing / metabolism
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Sarcoma, Ewing / pathology
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Structure-Activity Relationship
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Tumor Cells, Cultured
Substances
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4,7-dichloro-3-(2-(4-(dimethylamino)phenyl)-2-oxoethyl)-3-hydroxyindolin-2-one
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Aniline Compounds
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Antineoplastic Agents
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EWS-FLI fusion protein
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Indoles
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Oncogene Proteins, Fusion
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Proto-Oncogene Protein c-fli-1
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RNA-Binding Protein EWS
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Luciferases