Design, Synthesis, and Biology of Inhibitors of Neuronal Nitric Oxide Synthase

神经元一氧化氮合酶抑制剂的设计、合成和生物学

基本信息

批准号：
8810598
负责人：
Maris A Cinelli
金额：
$ 5.6万
依托单位：
NORTHWESTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8810598
关键词：
Active Sites Adverse effects Affinity Chromatography Alzheimer&apos s Disease Amidines Amines Aminoquinolines Arginine Binding Bioavailable Biological Biological Assay Biological Availability Biology Blood - brain barrier anatomy Brain Carboxylic Acids Caring Cell Membrane Permeability Cells Chronic Data Disease Docking Drug Kinetics Drug Targeting Enzyme Inhibition Enzymes Escherichia coli Evaluation Goals Halogens Heme Hemoglobin Huntington Disease Hydrogen Bonding Lead Literature Liver Microsomes Metabolic Migraine Modeling Modification Molecular Models Nerve Degeneration Neuraxis Neurons Nitric Oxide Synthase Type I Nitric Oxide Synthetase Inhibitor Palliative Care Parkinson Disease Pathology Pattern Penetration Permeability Pharmaceutical Preparations Phenethylamines Physiological Positioning Attribute Procedures Property Protein Isoforms Public Health Rattus Reporting Route Signaling Molecule Sodium Chloride Solubility Stroke Structure Symptoms System Testing Therapeutic Work alkalinity attenuation base design economic impact health economics improved inhibitor/antagonist mimetics molecular modeling monolayer oxetane public health relevance scaffold stability testing uptake

项目摘要

DESCRIPTION (provided by applicant): Although diseases characterized by neuronal damage and degeneration have an enormous public health and economic impact, treatment of these disorders is often limited to palliative care and slowing of symptom progression. Therefore, drugs that stop or slow neurodegeneration are highly desirable. One emerging target is neuronal nitric oxide synthase (nNOS), an enzyme that produces the signaling molecule NO. Although required for normal neuronal function, high levels of NO have been implicated in chronic neurodegenerative pathologies (such as Parkinson's disease) as well as stroke, migraines, and other disorders. Ergo, inhibition of this enzyme could be desirable for treatment of these diseases. Nonetheless, as most nNOS inhibitors mimic the natural substrate L-arginine, their therapeutic practicality is diminished by their excessive polarity and basicity, properties tat cause poor GI uptake and low blood-brain barrier permeability. Additionally, care must be taken to not inhibit the related NOS isoforms eNOS and iNOS, or dangerous side effects could result. The work detailed herein describes several strategies for the design and optimization of bioavailable nNOS inhibitors. First, preliminary data indicates that the N-benzylphenethylamine core is a scaffold that confers potent nNOS activity and ~100-fold isoform selectivity. In Aim 1, grafting a low-pKa heterocycle onto this scaffold via facile synthetic routes should result in a less basic arginine mimetic, and molecular modeling provides evidence that these compounds should bind in a manner similar to reported nNOS inhibitors. Additional optimizations will then be performed on compounds containing effective alternative heterocycles. In Aim 2a, incorporation of halogens and halogen- containing groups, a strategy that has proven effective at enhancing brain penetration for many classes of CNS drugs, will be performed. In Aim 2b, oxetane groups will be introduced into the phenethyl chain to decrease the high pKa of the secondary amines, a modification that is predicted to preserve the hydrogen-bonding capability of these amines without steric encumbrance. Finally, in Aim 3, nNOS and its isoforms will be expressed in E. coli and purified. Compounds will be assayed against the enzymes by the hemoglobin capture assay. In addition, select compounds will be tested in a cell-based nNOS assay, assayed for their metabolic stability, and tested for permeability in a Caco-2 model (to estimate both their GI and CNS uptake).

描述（由申请人提供）：虽然以神经元损伤和变性为特征的疾病具有巨大的公共健康和经济影响，但这些疾病的治疗通常仅限于姑息治疗和减缓症状进展。因此，非常需要能够阻止或减缓神经变性的药物。一个新兴靶标是神经元一氧化氮合酶 (nNOS)，这是一种产生信号分子 NO 的酶。虽然正常神经元功能需要高水平的一氧化氮，但高水平的一氧化氮与慢性神经退行性疾病（如帕金森病）以及中风、偏头痛和其他疾病有关。因此，抑制这种酶对于治疗这些疾病可能是理想的。尽管如此，由于大多数 nNOS 抑制剂模仿天然底物 L-精氨酸，其治疗实用性因其过度的极性和碱性而降低，这些特性会导致胃肠道吸收差和血脑屏障渗透性低。此外，必须注意不要抑制相关的 NOS 亚型 eNOS 和 iNOS，否则可能会导致危险的副作用。本文详述的工作描述了生物可利用的 nNOS 抑制剂的设计和优化的几种策略。首先，初步数据表明 N-苄基苯乙胺核心是一种支架，可赋予有效的 nNOS 活性和约 100 倍的异构体选择性。在目标 1 中，通过简单的合成途径将低 pKa 杂环接枝到该支架上应该会产生碱性较低的精氨酸模拟物，并且分子模型提供的证据表明这些化合物应该以类似于报道的 nNOS 抑制剂的方式结合。然后将对含有有效替代杂环的化合物进行额外的优化。在目标 2a 中，将采用卤素和含卤素基团的掺入策略，该策略已被证明可有效增强多种中枢神经系统药物的脑部渗透性。在目标 2b 中，氧杂环丁烷基团将被引入苯乙基链以降低仲胺的高 pKa，预计这种修饰将保留这些胺的氢键能力而没有空间阻碍。最后，在目标 3 中，nNOS 及其亚型将在大肠杆菌中表达并纯化。将通过血红蛋白捕获测定来针对酶来测定化合物。此外，选定的化合物将在基于细胞的 nNOS 测定中进行测试，测定其代谢稳定性，并在 Caco-2 模型中测试渗透性（以估计其 GI 和 CNS 摄取）。