Discovering small molecule ligands of vascular endothelial growth factor that block VEGF-KDR binding using label-free microarray-based assays

James P. Landry, Yiyan Fei, Xiangdong Zhu, Yaohuang Ke, Guoliang Yu, Pierre Lee

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

We present here a label-free microarray-based assay platform that we used to identify inhibitors of vascular endothelial growth factor (VEGF)-kinase-insertion domain receptor (KDR) binding. Supported by a combination of special ellipsometry-based optical detection and small molecule microarrays (SMM), this platform consists of three assays: (1) the first assay detects binding of a target protein with SMM and identifies ligands to the protein as inhibitor candidates; (2) the second assay detects binding of a receptor protein with identical SMM and subsequent binding of the target protein (a sandwich assay) to identify the ligands to the receptor protein that do not interfere with the target-receptor binding; (3) the third assay detects binding of the target protein to the receptor protein in the presence of the ligands of the target protein identified from the first assay, with the receptor protein immobilized to a solid surface through the ligands identified in the second assay, to yield dose-response curves. Using this platform, we screened 7,961 compounds from the National Cancer Institute and found 12 inhibitors to VEGF-KDR (VEGFR2) interactions with IC50 ranging from 0.3 to 60 μM. The inhibitory potency of these inhibitors found in the microarray-based assay was confirmed by their inhibition of VEGF-induced VEGFR2 phosphorylation in a cell-based assay.

Original languageEnglish (US)
Pages (from-to)326-332
Number of pages7
JournalAssay and Drug Development Technologies
Volume11
Issue number5
DOIs
StatePublished - Jun 1 2013

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Vascular Endothelial Growth Factor A
Phosphotransferases
Ligands
Proteins
Carrier Proteins
Immobilized Proteins
National Cancer Institute (U.S.)
Inhibitory Concentration 50
Phosphorylation

ASJC Scopus subject areas

  • Drug Discovery
  • Molecular Medicine
  • Medicine(all)

Cite this

Discovering small molecule ligands of vascular endothelial growth factor that block VEGF-KDR binding using label-free microarray-based assays. / Landry, James P.; Fei, Yiyan; Zhu, Xiangdong; Ke, Yaohuang; Yu, Guoliang; Lee, Pierre.

In: Assay and Drug Development Technologies, Vol. 11, No. 5, 01.06.2013, p. 326-332.

Research output: Contribution to journalArticle

Landry, James P. ; Fei, Yiyan ; Zhu, Xiangdong ; Ke, Yaohuang ; Yu, Guoliang ; Lee, Pierre. / Discovering small molecule ligands of vascular endothelial growth factor that block VEGF-KDR binding using label-free microarray-based assays. In: Assay and Drug Development Technologies. 2013 ; Vol. 11, No. 5. pp. 326-332.
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