PSYCHOTROPIC DRUGS AND RECEPTOR SENSITIVITY CHANGES

拟议的研究涉及多学科方法来开发 通过血脑屏障的新型精神治疗药物 刺激人类神经降压素受体。这些代理人可能是潜在的 治疗多种神经精神障碍的药物 精神分裂症和阿尔茨海默氏症。为了实现这一目标，我们 计划利用克隆的人神经降压素受体在 转染细胞系继续研究两种新的神经降压素 我们合成的受体激动剂。一个复合词代表一个 非肽类神经降压素合成的中间步骤 激动剂，是根据我们的分子模型设计的 神经降压素的计算机(8-13)。第二个是金刚烷衍生物 我们的一种新的神经降压素(8-13)肽类似物，似乎 穿透小白鼠的大脑。要了解更多关于结合位点(S)的信息 神经降压素和这些新化合物，我们计划进行研究- 人神经降压素受体的定向突变。这些研究 很可能是可行的，因为有了来自 我们通过聚合酶链法克隆了大鼠受体 反应(PCR)(来自人黑质CDNA文库，富含 神经降压素受体的来源)和人类文库的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个氨基酸中约有87%是可识别的 老鼠和人类受体之间的重叠。在获得完整的 序列，我们计划，就像我们对克隆的大鼠所做的那样 受体，研究人神经降压素受体在转基因细胞中的作用 排队。此外，由于我们对人类神经降压素的出色探针 受体，我们计划通过原位杂交来研究受体的表达。 正常人脑和阿尔茨海默病患者脑中的受体。至 鉴定表达的克隆人神经降压素受体，我们将做 神经降压素与膜的放射配基结合研究 从细胞中制备并测定神经降压素对第二代的影响 信使(肌醇磷酸和cAMP)由完整细胞合成。我们的 新的神经降压素受体激动剂将在这些测试中进行测试 表达的人神经降压素受体。最后，我们将在小鼠身上进行测试 这些化合物对戊巴比妥诱导的睡眠时间的影响， 对体温的影响，以及对体内止痛作用的研究 效果。

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.