Ketamine-Class Antidepressants in Vesicles

该项目研究氯胺酮及其代谢物和相关药物[氯胺酮类抗抑郁药 药物（KCAD），我们的术语]。亚麻醉剂量的氯胺酮产生抗抑郁作用， 通过未知的机制仅几个小时（2小时）。然而，更高的剂量会产生不利影响。 了解介导KCAD抗抑郁活性的机制是研究KCAD抗抑郁活性的重要一步。 药物开发的过程。这项研究计划的目标是从根本上改变我们的 了解这种快速的抗抑郁机制是如何工作的， 开发更强大和更安全的治疗方法。 KCAD作用的分子靶点未知。如果没有这样的知识， 应调查KCAD最集中的隔间内可能采取的行动。 我们检验了KCAD在大脑中的作用至少部分是由 KCAD在各种亚细胞区室（细胞器）中蓄积，包括突触 囊泡 目标1开发了一个新一代基因编码的KCAD“基于强度的氯胺酮传感荧光报告基因”（iKetSnFR）家族。这些将动态成像， 在亚细胞水平上定量KCAD的存在。目标1测量KCAD的时间进程 药物在细胞附近出现或消失后进入和退出各种细胞器。Aim 2测试 假设KCAD的抗抑郁机制涉及在管腔中蓄积 酸性囊泡，尤其是突触囊泡，随后是突触刺激诱导的释放 突触前末梢的KCAD。目的3检测KCAD诱导的神经递质释放， 突触前终端，采用下一代基因编码的生物传感器， 神经传递素这些实验还包括膜的电生理学研究， 突触特性 目标1、2和3的数据本身不会开发新的~2 h抗抑郁药。 但是，来自拟议实验的数据可以帮助理解KCAD如何发挥其 方面的影响.这些数据还将指导开发具有较少潜在侧的相关分子 作为治疗抑郁症的新的速效疗法。

The project studies ketamine, its metabolites, and related drugs [Ketamine-Class Antidepressant Drugs (KCADs), our term]. Sub-anesthetic doses of ketamine produce antidepressant effects in just a few hours (2 h) via unknown mechanism(s). However, higher doses have adverse effects. Understanding the mechanism mediating KCAD antidepressant activity is an important step in the process of drug development. This research program has the goal to fundamentally change our understanding of how this rapid antidepressant mechanism works and holds promise for development of more robust and safer treatments. The molecular target(s) of KCAD action are not known. In the absence of such knowledge, one should investigate possible actions in the compartments where KCADs are most concentrated. We test the hypothesis that the effects of KCADs in the brain are mediated, in least in part, by the accumulation of KCADs in various subcellular compartments (organelles), including synaptic vesicles. Aim 1 develops a family of next-generation genetically encoded “Intensity-based Ketamine-Sensing Fluorescent Reporters” (iKetSnFRs) for KCADs. These will dynamically image and quantify the presence of KCADs at sub-cellular levels. Aim 1 measures the time course of KCAD entry and exit from various organelles after the drugs appear or disappear near cells. Aim 2 tests the hypothesis that the antidepressant mechanism of KCADs involves accumulation in the lumen of acidic vesicles, especially in synaptic vesicles, followed by synaptic stimulation-induced release of KCAD from presynaptic terminals. Aim 3 detects KCAD-induced neurotransmitter release from presynaptic terminals, employing next generation genetically encoded biosensors for various neurotransmitters. The experiments also include electrophysiological studies of membrane and synaptic properties. The data from Aims 1, 2 and 3 will not in themselves develop a new ~2 h antidepressant drug. But the data from the proposed experiments can help to understand how KCADs exert their effects. The data will also guide the development of related molecules with fewer potential side effects as new fast-acting therapies for depression.