PSYCHOTROPIC DRUGS AND RECEPTOR SENSITIVITY CHANGES

拟议的研究涉及多学科的方法来开发 通过血脑屏障， 刺激人类神经降压素受体。 这些药物可能是潜在的 治疗各种神经精神疾病的药物， 精神分裂症和阿尔茨海默型痴呆。 为了实现这一目标，我们 计划使用稳定表达的克隆人神经降压素受体， 转染细胞系继续我们的工作，两个新的神经降压素 受体激动剂。 一种化合物代表 合成非肽类神经降压素的中间步骤 激动剂，并在我们的分子建模的基础上设计， 神经降压素的计算机（8-13）。 第二种是金刚烷衍生物 我们的一种新的神经降压素（8-13）肽类似物，似乎 穿透老鼠的大脑 要了解有关以下项的绑定位点的更多信息，请执行以下操作： 神经降压素和这些新的化合物，我们计划研究涉及网站- 人神经降压素受体的定向诱变。 这些研究 这可能是可行的，因为有了DNA序列信息， 克隆大鼠受体，我们已经获得了日期的聚合酶链反应 反应（PCR）（来自人黑质的cDNA文库，丰富的 神经降压素受体的来源）和人文库的cDNA筛选， 约90%的人神经降压素受体开放阅读框序列。 在氨基酸水平上，377个氨基酸的同源性约为87 大鼠和人类受体的重叠。 在获得整个 我们计划，就像我们对克隆大鼠所做的那样， 受体，以研究转染细胞中的人神经降压素受体 线 此外，我们的探针很好地检测了人类神经降压素 受体，我们计划通过原位杂交研究的表达， 在正常人脑和阿尔茨海默病脑中的受体。 到 表征表达的克隆人神经降压素受体，我们将做 [3 H]神经降压素和膜蛋白的放射性配体结合研究 从细胞制备和测量神经降压素的影响， 信使（磷酸肌醇和CAMP）由完整细胞合成。 我们 新的神经降压素受体激动剂将在这些试验中进行测试， 表达的人神经降压素受体。 最后，我们将在老鼠身上进行测试 这些化合物对戊巴比妥诱导的睡眠时间的影响， 对体温和镇痛的影响，以表征其体内 方面的影响.

The proposed research involves a multi-disciplinary approach to develop novel psychotherapeutic agents that pass the blood-brain barrier to stimulate the human neurotensin receptor. These agents may be potential drugs for treatment of the diverse neuropsychiatric disorders of schizophrenia and Alzheimer's type dementia. To achieve this goal, we plan to use the cloned human neurotensin receptor stably expressed in a transfected cell line to continue our work on two novel neurotensin receptor agonists that we have synthesized. One compound represent an intermediate step toward the synthesis of non-peptide neurotensin agonists and was designed on the basis of our molecular modeling by computer of neurotensin (8-13). The second, is an adamantane derivative of one of our novel neurotensin (8-13) peptide analogs and appears to penetrate the brain of mice. To learn more about the binding site(s) for neurotensin and these novel compounds, we plan studies involving site- directed mutagenesis of the human neurotensin receptor. These studies will likely be feasible since, with DNA sequence information from the cloned rat receptor, we have obtained to date by polymerase chain reaction (PCR) (from a CDNA library of human substantia nigra, a rich source of neurotensin receptors) and CDNA screening of a human library, about 90% of the human neurotensin receptor open reading frame sequence. At the amino acid level, there is about 87% identify in a 377 amino acid overlap between the rat and human receptors. After obtaining the entire sequence in a clone, we plan, as we have done with the cloned rat receptor, to study the human neurotensin receptor in a transfected cell line. In addition, with our excellent probe for the human neurotensin receptor, we plan to study by in situ hybridization the expression of the receptor in normal human brain and in Alzheimer's disease brains. To characterize the expressed cloned human neurotensin receptor, we will do radioligand binding studies with [3H]neurotensin and membranal preparations from cells and measure neurotensin's effects on second messenger (inositol phosphates and CAMP) synthesis by intact cells. Our novel neurotensin receptor agonists will be tested in these assays with the expressed human neurotensin receptor. Finally, we will test in mice the effects of these compounds on sleep time induced by pentobarbital, on body temperature, and on analgesia to characterize their in vivo effects.