In vitro pharmacology and In vivo neuroprotective effects of CMPI analogues in an optimized Zebrafish model of Parkinsons disease

帕金森病（PD）由多种潜在病理引起，包括α-突触核蛋白的积累， 含有路易体、多巴胺能神经元的损失和其他神经递质通路的干扰。 虽然使用前体L-DOAP增加多巴胺的产生似乎对减少PD症状有效， 首先，随着PD的进展，多巴胺能神经元数量的持续下降减少了 L-DOPA的有效性，以及控制PD症状的额外治疗变得必要。缺乏 靶向PD的潜在病理并阻止其进展的疾病修饰药物代表了一种重要的 我们的知识差距和未满足的临床需求。因此，本申请旨在启动早期实验 在一个连续的研究，这可能导致临床相关的积极变构的发展， 烟碱乙酰胆碱受体（nAChR）的调节剂（PAM），其已知具有神经保护性、抗- 炎性和抗炎性，因此有望开发治疗疾病的药物- 改性PD的性能。 在这个提议中，我们利用了一个主要药效团CMPI，我们已经确定它是一个 （α 4）3（β 2）2 nAChR选择性PAM及其类似物，以开发选择性nAChR PAM并提供 它们对PD的体内功效的概念证据。我们提出以下具体目标：1）确定 作为（α 4）3（β 2）2 nAChR PAM的具有高效力、功效和选择性的CMPI类似物; 2A）对CMPI类似物的优化， 用于体内帕金森氏病药物筛选的斑马鱼模型;和2B）nAChR PAM对帕金森氏病的作用的评估。 PD进展的行为、解剖和分子测量。 在实现这些特定目标后，我们期望以高亲和力和选择性结合的CMPI类似物 并增强激动剂诱导的（α 4）3（β 2）2 nAChR反应。预期结果将 还提供了体内证据，支持α 4 β 2 nAChR PAM作为一种新策略的药理学价值， 减缓与PD相关的运动和认知衰退的进展。拟议的研究项目还将 优化具有可测量行为的可靠斑马鱼PD样表型动物模型的条件， 解剖学和生物学读数，适用于高通量筛选新的治疗化合物， 警局该模型将有助于减少为未来动物和人类选择先导化合物的时间和成本 研究和促进新的治疗方法的发展，用于治疗运动症状和认知 与帕金森病相关的衰退。一旦这些药物成为临床相关的，进展将有 致力于有效的PD治疗和推进国立卫生研究院的使命。

Parkinson’s Disease (PD) results from multiple underlying pathologies, including accumulation of a-synuclein- containing Lewy bodies, loss of dopaminergic neurons, and perturbation of other neurotransmitter pathways. While increasing dopamine production using the precursor L-DOAP seems effective at reducing PD symptoms at first, the continuous decline in the number of dopaminergic neurons as PD progresses reduces the effectiveness of L-DOPA, and additional therapy to control the PD's symptoms becomes necessary. The lack of disease-modifying drugs that target PD's underlying pathologies and stop its progression represents a significant gap in our knowledge and unmet clinical need. Therefore, this application aims to initiate early step experiments in a continuum of research that is likely to lead to the development of clinically relevant positive allosteric modulators (PAMs) of nicotinic acetylcholine receptors (nAChRs) which are known to have neuroprotective, anti- inflammatory, and precognitive properties and thus hold promise for the development of drugs with disease- modifying properties for PD. In this proposal, we are taking advantage of a lead pharmacophore, CMPI, that we have identified as an (α4)3(β2)2 nAChR-selective PAM and its analogs to develop selective nAChR PAMs and to provide proof-of- concept evidence of their in vivo efficacy against PD. We propose the following specific aims: 1) Identification of CMPI analogs with high potency, efficacy, and selectivity as (α4)3(β2)2 nAChR PAMs; 2A) Optimization of a zebrafish model for in vivo Parkinson’s disease drug screening; and 2B) Assessment of nAChR PAMs effect on behavioral, anatomical, and molecular measures of PD progression. Upon accomplishing these specific aims, we expect that CMPI analogs that bind with high affinity and selectivity to and potentiate agonist-induced responses of (α4)3(β2)2 nAChR will be identified. The anticipated results will also provide in vivo evidence to support the pharmacological merit of α4β2 nAChR PAMs as a novel strategy to slow the progression of motor and cognitive decline associated with PD. The proposed research project will also optimize conditions for a reliable zebrafish PD-like phenotype animal model with measurable behavioral, anatomical, and biological readouts suitable for high throughput screening of novel therapeutic compounds for PD. This model will help reduce the time and cost of selecting lead compounds for future animal and human studies and facilitate the development of novel therapeutics for the treatment of motor symptoms and cognitive decline associated with Parkinson’s disease. Once such drugs become clinically relevant, progress will have been made toward effective PD treatment and advancing the National Institutes of Health mission.