Quantitative high-throughput screening using a live-cell cAMP assay identifies small-molecule Agonists of the TSH receptor

被引：48

作者：

Titus, Steve ^{[1
]}

Neumann, Susanne ^{[2
]}

Zheng, Wei ^{[1
]}

Southall, Noel ^{[1
]}

Michael, Sam ^{[1
]}

Klumpp, Carleen ^{[1
]}

Yasgar, Adam ^{[1
]}

Shinn, Paul ^{[1
]}

Thomas, Craig J. ^{[1
]}

Inglese, James ^{[1
]}

Gershengorn, Marvin C. ^{[2
]}

Austin, Christopher P. ^{[1
]}

机构：

[1] NHGRI, NIH, Chem Genom Ctr, Bethesda, MD 20892 USA

[2] NIDDK, Clin Endocrinol Branch, NIH, Bethesda, MD USA

来源：

JOURNAL OF BIOMOLECULAR SCREENING | 2008年 / 13卷 / 02期

关键词：

thyroid-stimulating hormone; TSH; TSHR; TSHR agonist; quantitative high-throughput screening; qHTS; HTS; probe identification; CNG; PubChem;

D O I：

10.1177/1087057107313786

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The thyroid-stimulating hormone (TSH; thyrotropin) receptor belongs to the glycoprotein hormone receptor subfamily of 7-transmembrane spanning receptors. TSH receptor (TSHR) is expressed mainly in thyroid follicular cells and is activated by TSH. which regulates the growth and function of thyroid follicular cells. Recombinant TSH is used in diagnostic screens for thyroid cancer. especially in patients after thyroid cancer surgery. Currently, no selective small-molecule agonists of the TSHR are available. To screen for novel TSHR agonists, the authors miniaturized a commercially available cell-based cyclic adenosine 3 ',5 ' monophosphate (cAMP) assay into a 1536-well plate format. This assay uses an HEK293 cell line stably transfected with the TSHR coupled to a cyclic nucleotide gated ion channel as a biosensor. From a quantitative high-throughput screen of 73,180 compounds in parallel with a parental cell line (without the TSHR), 276 primary active compounds were identified. The activities of the selected active compounds were further confirmed in an orthogonal homogeneous time-resolved fluorescence cAMP-based assay. Forty-nine compounds in several structural classes have been confirmed as the small-molecule TSHR agonists that will serve as a starting point for chemical optimization and studies of thyroid physiology in health and disease.

引用

页码：120 / 127

页数：8

共 18 条

[1] THE FUNCTIONAL EXPRESSION OF RECOMBINANT HUMAN THYROTROPIN RECEPTORS IN NONTHYROIDAL EUKARYOTIC CELLS PROVIDES EVIDENCE THAT HOMOLOGOUS DESENSITIZATION TO THYROTROPIN STIMULATION REQUIRES A CELL-SPECIFIC FACTOR [J].