Drug Transport at the CNS Barriers

中枢神经系统屏障的药物转运

• 批准号: 7939460
• 负责人: Joanne Wang
• 金额: $ 22.05万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7939460
• 关键词: 1-Methyl-4-phenylpyridinium; Adenosine; Affinity; Amantadine; Amino Acids; Amphetamines; Animal Model; Apical; Applications Grants; Biochemical; Biogenic Amines; Biological Process; Blood; Blood - brain barrier anatomy; Brain; Carrier Proteins; Cations; Cell membrane; Cell model; Charge; Computational Biology; Development; Dopamine; Drug Delivery Systems; Drug Kinetics; Drug Transport; Drug usage; Electrophysiology (science); Electrostatics; Epithelial Cells; Exposure to; Family; Funding; Gene Targeting; Genes; Goals; Homeostasis; Human; Immunohistochemistry; Kidney; Knowledge; Laboratories; Liver; Mediating; Membrane; Membrane Potentials; Methods; Molecular; Molecular Models; Names; Neuraxis; Neurotoxins; Neurotransmitters; Nicotine; Nucleoside Transporter; Nucleosides; Organic Cation Transporter; Pargyline; Pathway interactions; Permeability; Pharmaceutical Preparations; Pharmacodynamics; Physiological; Play; Poison; Progress Reports; Research; Rodent; Role; Sequence Homology; Serotonin; Site; Structure; Structure of choroid plexus; Substrate Interaction; Substrate Specificity; Techniques; Technology; Toxin; Transgenic Animals; Transgenic Organisms; Work; Xenobiotics; basolateral membrane; blood cerebrospinal fluid barrier; carrier mediated transport; in vivo; member; molecular modeling; monoamine; mouse model; novel; nucleoside analog; recombinase; social; solute; three-dimensional modeling

DESCRIPTION (provided by applicant): Organic cation (OC) transporters play important roles in the disposition and clearance of many endogenous and foreign OCs in the body. In contrast to the kidney and liver, little is known about OC elimination mechanisms in the brain. This is rather unfortunate because many CNS active compounds, including monoamine neurotransmitters (e.g. dopamine, serotonin), CNS drugs (e.g. amantadine, nicotine) and neurotoxins (e.g. 1-methyl-4-phenylpyridinium (MPP+)) are small hydrophilic OCs that rely on transporters to regulate their brain levels. This proposal focuses on plasma membrane monoamine transporter (PMAT), a novel brain OC transporter first cloned in our laboratory. While structurally related to the equilibrative nucleoside transporter family (SLC29), PMAT possesses a unique and surprisingly diverse substrate specificity, transporting structurally heterogeneous OCs such as biogenic amines, clinically used drugs and neurotoxins. In humans and rodents, PMAT is most abundantly expressed in the brain and highly concentrated in the blood-cerebrospinal fluid (CSF) barrier (i.e. BCSFB or choroid plexus). We hypothesized that PMAT is the principal OC transporter at the BCSFB and is responsible for removing a variety of endogenous and xenobiotic OCs from the brain. Three Specific Aims (SAs) have been proposed. SA1 is focused on elucidating the molecular mechanisms governing transporter-substrate interactions to explain the unique and versatile substrate specificity of PMAT. SA2 is focused on elucidating the transport mechanism of PMAT and developing a cellular model for OC flux at the BCSFB. Lastly, in SA3, we will construct and validate a transgenic animal model, which will allow a variety of mechanistic studies to reveal the in vivo action of PMAT. We will apply several experimental methods, ranging from molecular biochemical techniques, computational biology, electrophysiology, immunohistochemistry to transgenic approach, to elucidate the structure, function and biological significance of PMAT in clearing neurotransmitters, drugs and toxins in the brain. The proposed studies have broad implications in our understanding of normal and pathophysiological functions of the brain. Detailed knowledge of OC transporters at the CNS barriers can also help to explain and predict the pharmacokinetics and pharmacodynamics of OC drugs and toxins in the CNS compartment and aid in the development of new strategies for drug targeting to the brain.

描述（由申请人提供）：有机阳离子（OC）转运蛋白在体内许多内源性和外源性 OC 的处置和清除中发挥着重要作用。与肾脏和肝脏相比，人们对大脑中 OC 的消除机制知之甚少。这是相当不幸的，因为许多中枢神经系统活性化合物，包括单胺神经递质（例如多巴胺、血清素）、中枢神经系统药物（例如金刚烷胺、尼古丁）和神经毒素（例如1-甲基-4-苯基吡啶鎓（MPP+））都是小的亲水性有机化合物，依赖于转运蛋白来调节他们的大脑水平。该提案重点关注质膜单胺转运蛋白（PMAT），这是我们实验室首次克隆的一种新型脑 OC 转运蛋白。虽然在结构上与平衡核苷转运蛋白家族 (SLC29) 相关，但 PMAT 具有独特且令人惊讶的多样化底物特异性，可转运结构异质的 OC，例如生物胺、临床使用的药物和神经毒素。在人类和啮齿动物中，PMAT 在大脑中表达最丰富，并且高度集中在血脑脊液 (CSF) 屏障（即 BCSFB 或脉络丛）中。我们假设 PMAT 是 BCSFB 的主要 OC 转运蛋白，负责从大脑中去除各种内源性和外源性 OC。提出了三个具体目标（SA）。 SA1 专注于阐明控制转运蛋白-底物相互作用的分子机制，以解释 PMAT 独特且多功能的底物特异性。 SA2 致力于阐明 PMAT 的转运机制并开发 BCSFB OC 通量的细胞模型。最后，在SA3中，我们将构建并验证转基因动物模型，这将允许各种机制研究揭示PMAT的体内作用。我们将应用多种实验方法，从分子生化技术、计算生物学、电生理学、免疫组织化学到转基因方法，阐明PMAT在清除大脑中的神经递质、药物和毒素方面的结构、功能和生物学意义。拟议的研究对于我们理解大脑的正常和病理生理功能具有广泛的意义。对 CNS 屏障处 OC 转运蛋白的详细了解还有助于解释和预测 OC 药物和毒素在 CNS 区室中的药代动力学和药效学，并有助于开发针对大脑的药物靶向新策略。