Selective inhibition of nitric oxide synthase for multiple indications

选择性抑制一氧化氮合酶用于多种适应症

• 批准号: 10611534
• 负责人: RICHARD B SILVERMAN
• 金额: $ 42.96万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10611534
• 关键词: Alzheimer' s Disease; Antibiotics; Bacteria; Bacterial Infections; Biological Availability; Blood - brain barrier anatomy; Cell Line; Cerebral Palsy; Crystallography; Drug Delivery Systems; Generations; Grant; Growth; Hydrogen Peroxide; Melanoma Cell; Membrane; Metabolism; Modeling; Neoplasm Metastasis; Nerve Degeneration; Neurodegenerative Disorders; Nitric Oxide; Nitric Oxide Synthase; Nitric Oxide Synthase Type I; Nitric Oxide Synthetase Inhibitor; Oral; Oryctolagus cuniculus; Parkinson Disease; Peptides; Photoaffinity Labels; Property; Proteins; Proteomics; Resistance; Testing; Xenograft procedure; experimental study; inhibitor; melanocyte; melanoma; methicillin resistant Staphylococcus aureus; mouse model; oxidant stress; peptide drug; pharmacokinetics and pharmacodynamics; side effect

Project Summary This proposal is a continuation of R01 GM049725, “Selective Inhibition of Nitric Oxide Synthase for Multiple Indications”. Nitric oxide synthase (NOS) is a remarkable target, as we have found that neuronal nitric oxide synthase (nNOS) inhibitors are applicable to the potential treatment of neurodegenerative diseases (e.g., Parkinson's, Alzheimer's, cerebral palsy), bacterial infection, and melanoma. Inhibitors of nNOS block the excess NO that can cause degeneration of neurons. They also inhibit NO produced in melanocytes that leads to melanoma. Inhibitors of bacterial NOS (very similar to nNOS) can be used to protect antibiotics from bacterial degradation (i.e., resistance) and can be synergistic for bacterial growth inhibition by oxidant stress molecules, such as hydrogen peroxide. We have made outstanding progress on all three indications; two of which could not have progressed without the crystallography expertise of Thomas Poulos. In the last four years we have been able to shorten the syntheses of our inhibitors, make inhibitors that are membrane penetrable, are stabilized from metabolism, and are blood-brain barrier (BBB) penetrable with good oral bioavailability while maintaining excellent potency and nNOS selectivity. However, we have not yet gotten compounds that possess all of the desired properties. Consequently, this grant will allow us to identify compounds that have all of the desired pharmacodynamics and pharmacokinetic properties. We plan to optimize our nNOS-selective compounds for pharmacodynamics and pharmacokinetics and test in mouse models of Parkinson's and Alzheimer's diseases and in a rabbit model for cerebral palsy. We have identified several nNOS-selective compounds that inhibit the growth of bacteria, including MRSA, especially in combination with an antibiotic or hydrogen peroxide. We plan to make bacterial NOS (bNOS)-selective compounds to avoid potential side effects from inhibition of mammalian NOS. Also, there may be additional targets, other than bNOS with which our compounds interact; therefore, we will carry out protein pulldown experiments with a photoaffinity-labeled bNOS inhibitor and identify proteins to which it attaches with proteomics. This should clarify a more complete mechanism of action of bNOS inhibitors. Our best nNOS-selective inhibitors will also be tested by Dr. Sun Yang as inhibitors of a variety of melanoma cell lines and in a xenograft melanoma model for inhibition of melanoma growth and metastasis. Dr. Yang has identified a peptide carrier to target molecules to melanoma; we will conjugate active compounds to this peptide for drug delivery. We also will make a photoaffinity probe of an active inhibitor to identify targets.

项目概要 该提案是 R01 GM049725“选择性抑制一氧化氮 多种适应症的合酶”。一氧化氮合酶（NOS）是一个值得注意的目标，因为我们已经 发现神经元一氧化氮合酶（nNOS）抑制剂适用于潜在的治疗 神经退行性疾病（例如帕金森病、阿尔茨海默病、脑瘫）、细菌感染和 黑色素瘤。 nNOS 抑制剂可阻止过量的 NO 导致神经元退化。他们 还可以抑制黑色素细胞中产生的 NO，从而导致黑色素瘤。细菌 NOS 抑制剂（非常 类似于 nNOS）可用于保护抗生素免遭细菌降解（即耐药性）和 可以通过氧化应激分子（例如氢）协同抑制细菌生长 过氧化物。我们在所有三个指标上都取得了显着的进展；其中两个不可能有 在没有托马斯·普洛斯（Thomas Poulos）的晶体学专业知识的情况下取得了进展。在过去的四年里，我们 能够缩短抑制剂的合成，使抑制剂具有膜渗透性， 新陈代谢稳定，并且通过良好的口服可穿透血脑屏障 (BBB) 生物利用度，同时保持优异的效力和 nNOS 选择性。然而，我们还没有 获得具有所有所需特性的化合物。因此，这笔赠款将使我们能够 鉴定具有所有所需药效学和药代动力学特性的化合物。 我们计划优化我们的 nNOS 选择性化合物的药效学和 帕金森病和阿尔茨海默病小鼠模型以及兔子的药代动力学和测试 脑瘫模型。 我们已经鉴定出几种抑制细菌生长的 nNOS 选择性化合物， 包括 MRSA，尤其是与抗生素或过氧化氢联合使用。我们计划制作 细菌一氧化氮合酶（bNOS）选择性化合物，以避免抑制细菌一氧化氮合酶（bNOS）的潜在副作用 哺乳动物 NOS。此外，除了 bNOS 之外，我们的化合物还可能有其他靶点 相互影响;因此，我们将使用光亲和标记的 bNOS 进行蛋白质下拉实验 抑制剂并通过蛋白质组学鉴定其附着的蛋白质。这应该澄清更多 bNOS 抑制剂的完整作用机制。 我们最好的nNOS选择性抑制剂也将由孙杨博士作为多种抑制剂进行测试 黑色素瘤细胞系和异种移植黑色素瘤模型中抑制黑色素瘤生长和 转移。杨博士发现了一种肽载体，可以将分子靶向黑色素瘤；我们将 将活性化合物与该肽结合以进行药物输送。我们还将制作光亲和探针 活性抑制剂来识别目标。