Design of specific Molecular Probes for a Prostacyclin Receptor

前列环素受体特异性分子探针的设计

基本信息

批准号：
06558090
负责人：
SUZUKI Masaaki
金额：
$ 11.9万
依托单位：
Gifu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06558090/
关键词：
Prostacyclin Isocarbacyclin Receptor 15R-TIC 受容体探索子 APNIC 受容体タンパク質クローニング

项目摘要

Isocarbacyclin, a stable carbocyclic analog of prostacyclin, has efficiently been synthesized by combination of rationally designed organic reactions starting a universal intermediate readily obtained by our three-component coupling process. An efficient photoaffinity probe, APNIC,for the PGI_2 receptor (IP_1) has been elaborated by modification of isocarbacyclin. APNIC has proven to exhibit a strong and high specific affinity with the PGI_2 receptor protein (IC_<50>=3nM) in mastocytoma P-815 cells and act as a PGI_2 receptor agonist. The photoaffinity labeling experiment allowed for the first time the idenification of the PGI_2 receptor proteins of approximately 43 kDa (mastocytoma cells), 45 kDa (porcine platelets), and 52 kDa (human platelets) as glycoproteins. We discovered a novel prostacyclin (PGI_2) receptor (IP_2) expressed in the central nervous system (CNS) and succeeded in creating a stable ligand, (15R) -16-m-tolyl-17,18,19,20-tetranorisocarba- cyclin (15R-TIC), with high b … More inding affinity and selectivity for this receptor. 15R-TIC exhibited high binding affinity (IC_<50>=30nM) for a novel PGI_2 receptor (IP_2) in the thalamus but showed very weak binding (IC_<50>=1.2muM) for the PGI_2 receptor (IP_1) in the NTS (nucleus tractus solitarius). We examined the reaction of methyl iodide and tributylphenylstannane (excess), giving toluene, with the focus on the realization of rapid coupling applicable to short-lived ^<11>C radionuclide (t_<1/2>=20min). Although methylation by usual Stille reaction conditions failed. We accomplished this methylation in >90% yield using 40 equiv of tributylphenylstannane within 5 min at 60 ﾟC in the presence of bis(tri-o-tolylphosphine) palladium (0), Cu (I) salt, and K_2CO_3 in DMF.This new protocol provided a firm chemical basis for the synthesis of ^<11>CH_3-incorporated PET tracers and, actually, it was successfully applied to the synthesis of [^<11>C] 15R-TIC which realized clear imaging of the IP_2 receptor in the living monkey brain. Less

异卡巴环素是前列环素的稳定碳环类似物，通过合理设计的有机反应的组合，通过我们的三组分偶联过程轻松获得通用中间体，有效地合成了一种针对 PGI_2 受体 (IP_1) 的高效光亲和探针 APNIC。通过对异碳环素进行修饰而制成的药物已被证明与 PGI_2 受体蛋白具有强且高的特异性亲和力。 (IC_50>＝3nM)在肥大细胞瘤P-815细胞中并充当PGI_2受体激动剂光亲和标记实验首次允许鉴定约43kDa(肥大细胞瘤细胞)、45kDa()的PGI_2受体蛋白。猪血小板）和 52 kDa（人血小板）作为糖蛋白我们发现了一种新型前列环素。 (PGI_2) 受体 (IP_2) 在中枢神经系统 (CNS) 中表达，并成功创建稳定的配体 (15R) -16-m-tolyl-17,18,19,20-tetranorisocarbacyclin (15R-TIC) ，对该受体具有高亲和力和选择性，15R-TIC 对 a 表现出高结合亲和力 (IC_<50>=30nM)。丘脑中的新型 PGI_2 受体（IP_2），但与 NTS（孤束核）中的 PGI_2 受体（IP_1）的结合非常弱（IC_50>=1.2μM）。我们检查了甲基碘和三丁基苯基锡烷（过量）的反应。），给予甲苯，重点是实现适用于短命^<11>C的快速耦合放射性核素（t_<1/2>=20 分钟），但在双（三-）存在下，使用 40 当量的三丁基苯基锡烷在 5 分钟内完成了常规 Stille 反应条件的甲基化失败。邻甲苯基膦）钯 (0)、Cu (I) 盐和 K_2CO_3 的 DMF 溶液。这个新方案提供了为合成^<11>CH_3掺入的PET示踪剂奠定了坚实的化学基础，实际上，它已成功应用于[^<11>C]15R-TIC的合成，实现了活体猴子IP_2受体的清晰成像少脑