Analytical Methods for Investigating Peptide Transport

这些基于多肽的药物通常对所需受体表现出非常特异的靶向性。但是，在 为了发挥它们的治疗效果，它们还必须能够进入大脑并在酶的作用下 稳定。在这个方案中，强啡肽A(Dyn A)在血脑屏障(BBB)的运输和代谢 被调查过了。Dyn A是kappa阿片受体的天然配体。强啡肽及其类似物 代谢物已被研究用于治疗外周疼痛、药物成瘾和情感性 精神错乱。矛盾的是，Dyn A也被证明具有神经毒性。体内这种多肽的水平升高 中枢神经系统与神经病理性疼痛和阿尔茨海默病有关。 神经肽，如强啡肽，可以在体内发挥多种作用，包括神经递质， 神经调节剂和荷尔蒙。与小分子神经递质，如谷氨酸和 多巴胺，神经肽在释放后不被神经元重新吸收，而是在 胞外空间变成更小的多肽片段。这些碎片可以直接影响 周围区域的细胞外空间，穿过血脑屏障，在远离的地方发挥作用 从他们最初发布的网站上。 这项提议有三个主要目标。第一个是研究药物的转运和代谢。 Dyn A1-17，其代谢物，以及BBB中的合成类似物.通过以下方式获取的信息 实验将导致对运输机制的更好的理解，并将使有可能 生产针对kappa阿片受体的更好的治疗药物。二是发展以分离为基础的 可用于监测强啡肽及其代谢物对神经递质影响的传感器 在CMS中发布。第三个目标是开发一种基于分离的传感器，可以放在动物身上 因此，神经递质和行为的测量可以同时进行，使用觉醒， 自由漫游的动物。这些技术将被用于研究Dyn A的药理作用 血脑屏障和中枢神经系统。这里开发的方法学将普遍适用于所有神经化学物质。 需要测量血脑屏障转运或神经递质的研究，特别是 那些监控行为也值得关注的案例。 意义：强啡肽是一种重要的神经肽，与疼痛、可卡因成瘾、 抑郁症和阿尔茨海默氏症。更好地了解这种多肽的运输和代谢 BBB将协助科学家为这些疾病提供更好的治疗方法。分析性的 这项提案中开发的方法也将对参与中枢神经系统研究的科学家具有普遍意义。

These peptide-based drugs normally exhibit very specific targeting to the desired receptor. However, in order to exert their therapeutic effects, they must also be able to cross into the brain and be enzymatically stable. In this proposal, the transport and metabolism of dynorphin A (Dyn A) at the blood-brain barrier (BBB) is investigated. Dyn A is a natural ligand of the kappa opioid receptor. Analogs of dynorphin and its metabolites have been investigated for the treatment of peripheral pain, drug addiction, and affective disorders. Paradoxically, Dyn A has also been shown to be neurotoxic. Elevated levels of this peptide in the CNS has been associated with neuropathic pain and Alzheimer's disease. Neuropeptides, such as dynorphin, can play multiple roles in vivo including that of a neurotransmitter, neuromodulator, and hormone. In contrast to small molecule neurotransmitters such as glutamate and dopamine, neuropeptides are not taken back up by neurons following their release but are metabolized in the extracellular space into smaller peptide fragments. These fragments can directly affect the cells in the extracellular space in the surrounding area, pass through the BBB, and exert effects at sites far removed from the site of their initial release. There are three main goals of this proposal. The first is to investigate the transport and metabolism of Dyn A 1-17, its metabolites, and synthetic analogs across the BBB. Information acquired through these experiments will lead to a better understanding of the mechanism of transport and will make it possible to produce better therapeutics targeted at the kappa opioid receptor. The second is to develop separationbased sensors that can be used to monitor the effects of dynorphin and its metabolites on neurotransmitter release in the CMS. The third objective is to develop a separation-based sensor that can be placed onanimal so that measurements of neurotransmitters and behavior can be made simultaneously, using awake, freely roaming animals. These techniques will be employed to study the pharmacological actions of Dyn A at the BBB and in the CNS. The methodology developed here will be generally applicable to all neurochemical studies in which the measurement of blood brain barrier transport or neurotransmitters is desired, especially those cases where monitoring behaviour is also of interest. Significance: Dynorphin is an important neuropeptide that is involved in pain, cocaine addiction, depression, and Alzheimer's disease. A better understanding of the transport and metabolism of this peptide across the BBB will assist scientists in producing better treatments for these diseases. The analytical methods developed in this proposal will also be of general utility to scientists involved in CNS research.