PSYCHOTROPIC DRUGS AND RECEPTOR SENSITIVITY CHANGES

精神药物和受体敏感性变化

• 批准号: 2244170
• 负责人: ELLIOTT RICHELSON
• 金额: $ 32.43万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-06-01 至 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2244170
• 关键词: Alzheimer's disease; amantadine; blood brain barrier; brain mapping; chemical binding; computer simulation; drug design /synthesis /production; human tissue; in situ hybridization; laboratory rat; molecular cloning; neuropeptide receptor; neurotensin; nucleic acid sequence; peptide analog; pharmacokinetics; polymerase chain reaction; psychopharmacology; psychotropic drugs; receptor expression; schizophrenia; second messengers; site directed mutagenesis; stimulant /agonist; transfection

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.

拟议的研究涉及多学科方法来开发 新型精神治疗药物可通过血脑屏障 刺激人类神经降压素受体。 这些药物可能有潜力 治疗多种神经精神疾病的药物 精神分裂症和阿尔茨海默氏型痴呆。 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 转染细胞系以继续我们对两种新型神经降压素的研究 我们合成的受体激动剂。 一种化合物代表一种 合成非肽神经降压素的中间步骤 激动剂，是根据我们的分子模型设计的 神经降压素计算机 (8-13)。 第二种是金刚烷衍生物 我们的一种新型神经降压素 (8-13) 肽类似物，似乎 穿透老鼠的大脑。 了解有关结合位点的更多信息 神经降压素和这些新化合物，我们计划进行涉及位点的研究 人类神经降压素受体的定向诱变。 这些研究 可能是可行的，因为有了来自 克隆大鼠受体，我们迄今为止通过聚合酶链获得 反应 (PCR)（来自人类黑质的 cDNA 文库，黑质是一种丰富的 神经降压素受体的来源）和人类文库的 cDNA 筛选， 约90%的人类神经降压素受体开放阅读框序列。 在氨基酸水平上，377个氨基酸的同一性约为87% 大鼠和人类受体之间的重叠。 获取全部后 我们计划克隆中的序列，就像我们对克隆大鼠所做的那样 受体，研究转染细胞中的人类神经降压素受体 线。 此外，凭借我们出色的人类神经降压素探针 受体，我们计划通过原位杂交研究该受体的表达 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，我们会做 [3H]神经降压素和膜的放射性配体结合研究 细胞制剂并测量神经降压素对第二个细胞的影响 完整细胞合成信使（磷酸肌醇和 CAMP）。 我们的 新型神经降压素受体激动剂将在这些测定中进行测试 表达的人类神经降压素受体。 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， 体温和镇痛来表征它们的体内 影响。