title: Synthesis and structure-activity relationship studies of small molecule disruptors of EWS-FLI1 interactions in Ewing's sarcoma.
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).
authors.affiliation: Center for Drug Discovery, Georgetown University Medical Center , New Research Building EP07, 3970 Reservoir Road, NW, Washington, D.C. 20057, United States.
authors.forename: Perrer N
authors.initials: PN
authors.lastname: Tosso
authors.forename: Yali
authors.initials: Y
authors.lastname: Kong
authors.forename: Lauren
authors.initials: L
authors.lastname: Scher
authors.forename: Ryan
authors.initials: R
authors.lastname: Cummins
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Schneider
authors.forename: Said
authors.initials: S
authors.lastname: Rahim
authors.forename: K Travis
authors.initials: KT
authors.lastname: Holman
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Toretsky
authors.forename: Kan
authors.initials: K
authors.lastname: Wang
authors.forename: Aykut
authors.initials: A
authors.lastname: Üren
authors.forename: Milton L
authors.initials: ML
authors.lastname: Brown
mesh_terms: D000814:Aniline Compounds / Q000138:chemical synthesis / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000494:pharmacology*; D001859:Bone Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D049109:Cell Proliferation / Q000187:drug effects; D018360:Crystallography, X-Ray; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006801:Humans; D007211:Indoles / Q000138:chemical synthesis / Q000494:pharmacology*; D008156:Luciferases / Q000378:metabolism; D008958:Models, Molecular; D015394:Molecular Structure; D015514:Oncogene Proteins, Fusion / Q000037:antagonists & inhibitors* / Q000378:metabolism; D051789:Proto-Oncogene Protein c-fli-1 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D034802:RNA-Binding Protein EWS / Q000037:antagonists & inhibitors* / Q000378:metabolism; D012512:Sarcoma, Ewing / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
chemical_list: C000595864:4,7-dichloro-3-(2-(4-(dimethylamino)phenyl)-2-oxoethyl)-3-hydroxyindolin-2-one; D000814:Aniline Compounds; D000970:Antineoplastic Agents; C106031:EWS-FLI fusion protein; D007211:Indoles; D015514:Oncogene Proteins, Fusion; D051789:Proto-Oncogene Protein c-fli-1; D034802:RNA-Binding Protein EWS; D008156:Luciferases