Neumann Susanne, Huang Wenwei, Titus Steve, Krause Gerd, Kleinau Gunnar, Alberobello Anna Teresa, Zheng Wei, Southall Noel T, Inglese James, Austin Christopher P, Celi Francesco S, Gavrilova Oksana, Thomas Craig J, Raaka Bruce M, Gershengorn Marvin C
Clinical Endocrinology Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
Proc Natl Acad Sci U S A. 2009 Jul 28;106(30):12471-6. doi: 10.1073/pnas.0904506106. Epub 2009 Jul 10.
Seven-transmembrane-spanning receptors (7TMRs) are prominent drug targets. However, small-molecule ligands for 7-transmembrane-spanning receptors for which the natural ligands are large, heterodimeric glycoprotein hormones, like thyroid-stimulating hormone (TSH; thyrotropin), have only recently been reported, and none are approved for human use. We have used quantitative high-throughput screening to identify a small-molecule TSH receptor (TSHR) agonist that was modified to produce a second agonist with increased potency. We show that these agonists are highly selective for human TSHR versus other glycoprotein hormone receptors and interact with the receptor's serpentine domain. A binding pocket within the transmembrane domain was defined by docking into a TSHR homology model and was supported by site-directed mutagenesis. In primary cultures of human thyrocytes, both TSH and the agonists increase mRNA levels for thyroglobulin, thyroperoxidase, sodium iodide symporter, and deiodinase type 2, and deiodinase type 2 enzyme activity. Moreover, oral administration of the agonist stimulated thyroid function in mice, resulting in increased serum thyroxine and thyroidal radioiodide uptake. Thus, we discovered a small molecule that activates human TSHR in vitro, is orally active in mice, and could be a lead for development of drugs to use in place of recombinant human TSH in patients with thyroid cancer.
七跨膜受体(7TMRs)是重要的药物靶点。然而,对于天然配体为大型异二聚体糖蛋白激素(如促甲状腺激素(TSH;促甲状腺素))的七跨膜受体,其小分子配体直到最近才被报道,且尚无被批准用于人类的药物。我们利用定量高通量筛选鉴定出一种小分子促甲状腺激素受体(TSHR)激动剂,并对其进行修饰以产生效力增强的第二种激动剂。我们表明,这些激动剂对人TSHR具有高度选择性,与其他糖蛋白激素受体相比,它们与受体的蛇形结构域相互作用。通过对接TSHR同源模型确定了跨膜结构域内的一个结合口袋,定点诱变也证实了这一点。在人甲状腺细胞原代培养中,TSH和激动剂均可提高甲状腺球蛋白、甲状腺过氧化物酶、钠碘同向转运体和2型脱碘酶的mRNA水平以及2型脱碘酶的酶活性。此外,口服该激动剂可刺激小鼠的甲状腺功能,导致血清甲状腺素水平升高和甲状腺放射性碘摄取增加。因此,我们发现了一种在体外可激活人TSHR、在小鼠体内具有口服活性的小分子,它可能成为开发用于替代甲状腺癌患者重组人TSH的药物的先导化合物。