Nagasaki Hiroshi, Chung Shinjae, Dooley Colette T, Wang Zhiwei, Li Chunying, Saito Yumiko, Clark Stewart D, Houghten Richard A, Civelli Olivier
Dept. of Metabolic Medicine, School of Medicine, Nagoya University, Nagoya, Japan.
Eur J Pharmacol. 2009 Jan 14;602(2-3):194-202. doi: 10.1016/j.ejphar.2008.10.068. Epub 2008 Nov 17.
Melanin-concentrating hormone (MCH) is a neuropeptide that exhibits potent orexigenic activity. In rodents, it exerts its actions by interacting with one receptor, MCH(1) receptor which is expressed in many parts of the central nervous system (CNS). To study the physiological implications of the MCH system, we need to be able to block it locally and acutely. This necessitates the use of MCH(1) receptor antagonists. While MCH(1) receptor antagonists have been previously reported, they are mainly not accessible to academic research. We apply here a strategy that leads to the isolation of a high affinity and selective MCH(1) receptor antagonist amenable to in vivo analyses without further chemical modifications. This antagonist, TPI 1361-17, was identified through the screening of multiple non-peptide positional scanning synthetic combinatorial libraries (PS-SCL) totaling more than eight hundred thousand compounds in conditions that allow for the identification of only high-affinity compounds. TPI 1361-17 exhibited an IC(50) value of 6.1 nM for inhibition of 1 nM MCH-induced Ca(2+) mobilization and completely displaced the binding of [(125)I] MCH to rat MCH(1) receptor. TPI 1361-17 was found specific, having no affinity for a variety of other G-protein coupled receptors and channels. TPI 1361-17 was found active in vivo since it blocked MCH-induced food intake by 75%. Our results indicate that TPI 1361-17 is a novel and selective MCH(1) receptor antagonist and is an effective tool to study the physiological functions of the MCH system. These results also illustrate the successful application of combinatorial library screening to identify specific surrogate antagonists in an academic setting.
黑色素聚集激素(MCH)是一种具有强大促食欲活性的神经肽。在啮齿动物中,它通过与一种受体——MCH(1)受体相互作用来发挥作用,该受体在中枢神经系统(CNS)的许多部位都有表达。为了研究MCH系统的生理意义,我们需要能够在局部和急性条件下阻断它。这就需要使用MCH(1)受体拮抗剂。虽然之前已经报道过MCH(1)受体拮抗剂,但学术研究人员主要无法获得这些拮抗剂。我们在此应用了一种策略,该策略能够分离出一种高亲和力和选择性的MCH(1)受体拮抗剂,无需进一步化学修饰即可用于体内分析。这种拮抗剂TPI 1361 - 17是通过筛选多个非肽位置扫描合成组合文库(PS - SCL)鉴定出来的,这些文库总共包含超过八十万个化合物,筛选条件能够仅鉴定出高亲和力化合物。TPI 1361 - 17对1 nM MCH诱导的Ca(2+)动员的抑制作用的IC(50)值为6.1 nM,并能完全取代[(125)I] MCH与大鼠MCH(1)受体的结合。发现TPI 1361 - 17具有特异性,对多种其他G蛋白偶联受体和通道没有亲和力。发现TPI 1361 - 17在体内具有活性,因为它能将MCH诱导的食物摄入量减少75%。我们的结果表明,TPI 1361 - 17是一种新型的选择性MCH(1)受体拮抗剂,是研究MCH系统生理功能的有效工具。这些结果还说明了组合文库筛选在学术环境中成功应用于鉴定特定替代拮抗剂。