CHEMICAL STRUCTURE--HYPOTHALAMIC NA+/K+ ATPASE INHIBITOR

化学结构--下丘脑NA /K ATP酶抑制剂

• 批准号: 2750414
• 负责人: Garner Tripp Haupert
• 金额: $ 40.99万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-04-01 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2750414
• 关键词: cardiac glycosides; chemical structure function; circular dichroism; enzyme inhibitors; enzyme mechanism; hypothalamic hormones; immunoaffinity chromatography; monoclonal antibody; ouabain; sodium potassium exchanging ATPase

DESCRIPTION: Hypothalamic inhibitory factor, HIF, is felt to be a mammalian analog of the digitalis glycosides with biologic and biochemical properties consistent with physiologic regulation of the mammalian sodium pump in cardiovascular, renal and nervous system tissues in vivo. Disregulation of the inhibitor-Na,K-ATPase system has been linked to the pathogenesis of human essential hypertension. Structural analysis of pure HIF showed it to be an isomer, differing in the steroid moiety, of the plant glycoside, ouabain. Recently, an ouabain-like compound (OLC) isolated from human plasma and felt to be indistinguishable from plant ouabain has been restudied and found to be different from ouabain but identical to HIF, further supporting the notion that a unique molecule may specifically regulate the human Na,K-ATPase. The simplest explanation consistent with our spectroscopic data was that HIF differed from ouabain as a position isomer rhamnoside. We synthesized eight of these peracylated analogs and obtained their CD spectra. None of them showed weak CD curves (nearly zero) which is the signature of pentanaphthoyl HIF and OLC, nor did calculated CD spectra of remaining position isomers, with the possible exception of the C5 rhamnoside. With the likelihood of a more complex difference in the steroid moiety, an affinity purification method was developed using a monoclonal anti-digoxin antibody which resulted in high yields of pure HIF, making complete structural assignment of the endogenous Na,K-ATPase inhibitor achievable. Development of monoclonal antibodies to characterize HIF and create a diagnostic assay is newly proposed.

描述：下丘脑抑制因子（Hypothalamic inhibitory factor，HIF）是哺乳动物的一种神经递质， 具有生物学和生物化学性质的洋地黄苷类似物 与哺乳动物钠泵的生理调节一致， 心血管、肾脏和神经系统组织。 的失调 线粒体-Na，K-ATP酶系统已被链接到的发病机制 人类原发性高血压 纯HIF的结构分析表明， 是植物糖苷的异构体，在甾体部分上不同， 哇巴因 近年来，从人体组织中分离出一种类哇巴因化合物（OLC）， 血浆和感觉与植物哇巴因难以区分， 重新研究发现与哇巴因不同但与HIF相同， 进一步支持了一种独特的分子可以特异性地 调节人体Na，K-ATP酶。 最简单的解释是 我们的光谱数据是HIF不同于哇巴因， 异构体鼠李糖苷。 我们合成了八个这样的过酰化类似物， 获得了它们的CD光谱。 没有一个显示弱CD曲线（几乎为零） 这是戊萘酰HIF和OLC的特征，计算的CD也没有 其余位置异构体的光谱，可能的例外是C5 鼠李糖苷 类固醇中可能存在更复杂的差异 部分，使用单克隆抗体开发亲和纯化方法。 抗地高辛抗体导致高产量的纯HIF， 内源性Na，K-ATP酶抑制剂的完整结构归属 可实现的。 表征HIF和HIF-1 α的单克隆抗体的开发 新提出了创建诊断测定。