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

Isocarbacyclin是前列环蛋白的稳定羰基蛋白类似物，通过合理设计的有机反应的组合有效地合成了我们的三组分耦合过程，从而启动了通用中间体。对于PGI_2受体（IP_1）的有效光性探针已通过修饰异含酰胺激素详细阐述。事实证明，APNIC与PGI_2受体蛋白（IC_ <50> = 3nm）在肥大细胞瘤P-815细胞中表现出很强的特异性亲和力，并充当PGI_2受体激动剂。光性标记实验首次允许鉴定约43 kDa（肥大细胞），45 kDa（猪血小板）和52 kDa（人血型）的PGI_2受体蛋白作为糖蛋白。我们发现了一种新型的前列环蛋白（PGI_2）受体（IP_2），该受体（IP_2）在中枢神经系统（CNS）中表达，并成功地创建了稳定的配体（15R）-16-m-tolyl-17,18,19,19,20-四甲状腺果（15R），cyclin（15R）（15R-TIC），具有高度的亲身和选择性，可用于此受体和选择性。 15R-TIC暴露的高结合亲和力（IC_ <50> = 30nm）对于丘脑中的新型PGI_2受体（IP_2），但在NTS（nucleus tractus silesarius）中显示了PGI_2受体（IP_1）的非常弱的结合（IC_ <50> = 1.2MUM）。我们检查了碘化甲基甲基苯基甲烷（过量）的反应，给出了甲苯，重点是实现适用于短寿命 ^<11> c radioclide（T_ <1/2> = 20min）的快速耦合。尽管通常的反应条件甲基化失败了。我们在60°C的5分钟内使用40等等价在60°C的情况下，在BIS（Tri-O-Tolylyl膦）钯（0）（0），Cu（i）盐和K_2CO_3的情况下，在60°C的5分钟内，我们在60°C的5分钟内实现了这种甲基化，在60°C的甲基化中实现了这种甲基化。应用于[^<11> c] 15R-TIC的合成，该图清楚地意识到活着的猴子大脑中IP_2受体的清晰成像。较少的