Ketamine-Class Antidepressants in Vesicles

该项目研究氯胺酮、其代谢物和相关药物(氯胺酮类抗抑郁药 药物(KCADS)，我们的术语]。亚麻醉剂量的氯胺酮在脑内产生抗抑郁作用 只有几个小时(2小时)，机制未知(S)。然而，更高的剂量会产生不良影响。 了解调节KCAD抗抑郁活性的机制是在 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 了解这种快速抗抑郁机制是如何起作用的，并有望 开发更强大和更安全的治疗方法。 KCAD作用的分子靶点(S)尚不清楚。在缺乏这种知识的情况下，一个人 应调查在KCAD最集中的隔间可能采取的行动。 我们检验了这样一个假设，即KCAD在大脑中的作用至少部分是通过 KCAD在包括突触在内的各种亚细胞室(细胞器)中的积聚 水泡。 目的1为KCADS开发下一代基因编码的“基于强度的氯胺酮敏感荧光报告”(IKetSnFR)家族。它们将动态地成像和 在亚细胞水平上量化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，从各种细胞器进入和离开。AIM 2测试 KCADS的抗抑郁机制涉及腔内蓄积的假说 酸性小泡，特别是在突触小泡中，随后突触刺激诱导释放 来自突触前终末的KCAD。AIM 3检测KCAD诱导的神经递质释放 突触前终端，采用下一代遗传编码生物传感器用于各种 神经递质。这些实验还包括膜的电生理学研究和 突触特性。 来自AIMS 1、2和3的数据本身不会开发出新的~2小时抗抑郁药物。 但是，来自拟议实验的数据可以帮助理解KCAD如何发挥其作用 效果。这些数据还将指导潜在侧向较少的相关分子的发展 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.