Human Brain CYP P450s and Psychoactive Drug Metabolism

人脑 CYP P450 与精神活性药物代谢

基本信息

批准号：
6948530
负责人：
HENRY W STROBEL
金额：
$ 37.14万
依托单位：
UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-07-01 至 2008-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6948530
关键词：
alprazolam brain complementary DNA cytochrome P450 drug metabolism enzyme activity fluoxetine gene expression haloperidol imipramine immunocytochemistry in situ hybridization methylphenidate microsomes polymerase chain reaction psychotropic drugs western blottings

项目摘要

DESCRIPTION (provided by applicant): A number of cytochromes P450 expressed in liver have also been shown to be expressed in human and rat brain using techniques of in situ hybridization and immunohistochemical detection. Moreover, using human and rat brain microsomes, the P450-dependent metabolism of drugs and stercids has been demonstrated showing that brain cytochromes P450 are catalytically active. Recently a unique CYP2D form has been identified as expressed only in the brain of 9 out of 20 cadaveric donors examined. This CYP2D form, cloned and expressed in Neuro 2A cells, has a unique catalytic activity, converting codeine exclusively to morphine, thereby demonstrating not only unique brain specific localization, but also, a specific functional activity. This proposal will focus on the definition of expression of this and other P450 forms uniquely expressed in human brain. This objective will be accomplished by defining the regional distribution of such forms in brain as a function of gender, age and ethnicity as well as by definition of the catalytic capacities of these expressed purified P450s towards a panel of neuroactive psychoactive drugs clinically utilized to treat brain disorders. The drug panel will include the antidepressants fluoxetine and imipramine the neuroleptics chlorpromazine and haloperidol, the antianxiety drug alprazolam, the antihyperactivity drug methylphenidate, the analgesic ethylmorphine and the amphetamine benzphetamine. We will define the catalytic activities of these drugs in human brain microsomes. We will define the regional expression of these novel forms using QRTPCR with RNA isolated from brain regions as well as by in situ hybridization techniques. We will express the cloned unique forms in heterologous expression systems such as Neuro 2A, COS 7, E. coli and yeast cells in order to assess the catalytic activities of the expressed protein in comparison with brain microsomes. We will examine psychoactive substrate binding of the expressed unique proteins versus wild type forms. These approaches at various levels will enable us to define CYP P450 forms expressed uniquely in brain and assess their functional roles in metabolizing psychoactive drugs.

描述（由申请人提供）：使用原位杂交和免疫组织化学检测的技术在人和大鼠脑中表达了许多在肝脏中表达的细胞色素P450。此外，使用人和大鼠脑微粒体，已经证明了药物和刺激性的P450依赖性代谢，表明脑细胞染料P450是催化活性的。最近，仅在检查的20个尸体供体中的9个大脑中，已确定了一种独特的CYP2D形式。这种CYP2D形式在Neuro 2a细胞中克隆和表达，具有独特的催化活性，将可待因专门转换为吗啡，因此不仅证明了独特的大脑特定定位，而且还展示了特定的功能活性。该提议将重点介绍该表达的定义和其他P450在人脑中唯一表达的形式。该目标将通过定义大脑中这种形式的区域分布来实现，这是性别，年龄和种族的函数，以及通过定义这些表达纯化的P450的催化能力朝着一组神经活性的精神活性药物临床利用来治疗脑部疾病。该药物面板将包括抗抑郁药氟西汀和丙咪嗪神经蛋白酶氯丙嗪和氟哌啶醇，抗焦虑药物阿普唑仑，抗耐耐药性药物哌醋甲甲化物，镇痛乙基乙基乙基甲甲基和苯丙胺苯丙胺苯丙胺。我们将定义这些药物在人脑微粒体中的催化活性。我们将使用QRTPCR和从脑区域分离的RNA以及通过原位杂交技术来定义这些新型形式的区域表达。我们将在异源表达系统（例如Neuro 2a，cos 7，大肠杆菌和酵母细胞）等异源表达系统中表达克隆的独特形式，以评估与脑微粒体相比，评估表达蛋白的催化活性。我们将检查表达独特蛋白与野生型形式的精神活性底物结合。这些方法在各个层面上都将使我们能够定义CYP P450形式在大脑中唯一表达，并评估其在代谢精神药物中的功能作用。