Design And Development Of Experimental Therapeutics

实验疗法的设计和开发

• 批准号: 7732195
• 负责人: Nigel H. Greig
• 金额: $ 94.34万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7732195
• 关键词: 5' Untranslated Regions; Abeta synthesis; Acetylcholine; Acetylcholinesterase; Acetylcholinesterase Inhibitors; Acute; Affect; Aging; Alzheimer' s Disease; Amyloid beta-Protein; Amyotrophic Lateral Sclerosis; Animal Model; Apoptosis; Binding; Biochemical; Biochemistry and Pharmacology Cancer Activity; Biological; Brain; Brain region; Butyrylcholinesterase; Cell Death; Cells; Chemical Warfare; Chemistry; Cholinergic Agents; Cholinesterase Inhibitors; Cholinesterases; Chronic; Chronic Disease; Clinic; Clinical; Clinical Research; Clinical Trials; Clinical assessments; Cognition; Collaborations; Coupled; Craniocerebral Trauma; Cultured Cells; Development; Diabetes Mellitus; Disease; Dose; Drug Addiction; Drug Design; Drug Exposure; Drug Kinetics; Drug effect disorder; Elderly; Enzyme Inhibition; Enzymes; Evaluation; Event; Extramural Activities; Food; Frequencies; Functional disorder; GLP-I receptor; Glucose; Goals; Half-Life; Health; Hydrolysis; Individual; Industry; Inflammation; Japan; Kinetics; Laboratories; Mediating; Messenger RNA; Metabolism; Modeling; Molecular; Myasthenia Gravis; Nerve Degeneration; Nervous system structure; Neurodegenerative Disorders; Neuroglia; Neurons; Neurotransmitters; Non-Insulin-Dependent Diabetes Mellitus; Oxazoles; Parkinson Disease; Peptides; Peripheral Nerves; Peripheral Nervous System Diseases; Pharmaceutical Preparations; Phenylcarbamates; Physostigmine; Plasma; Process; Property; Protein Precursors; Proteins; Research; Rodent; Role; Scientist; Series; Stroke; Structure of beta Cell of islet; Symptoms; System; Systemic disease; TP53 gene; Testing; Thalidomide; Therapeutic; Time; Toxic effect; Transcriptional Activation; Translating; Translations; Traumatic Brain Injury; Tumor Necrosis Factor-alpha; United States National Institutes of Health; Up-Regulation; Variant; Vertebral column; X-Ray Crystallography; analog; base; cholinergic; cytokine; design; drug development; drug metabolism; exenatide; glucagon-like peptide 1; human TNF protein; in vivo; inhibitor/antagonist; insulin secretion; interest; neuroprotection; novel; phenserine; pre-clinical; preclinical study; prevent; prophylactic; response; therapy development; tissue culture; tolserine; tool; transcription factor; tumor necrosis factor-alpha inhibitor

1. Alzheimers Disease: Three series of agents are being developed to treat AD. Selective inhibitors of acetylcholinesterase (AChE), of butyrylcholinesterase (BChE), and of amyloid-beta peptide (Abeta) production. 1.1. Cholinesterase inhibitors: Compounds were developed to optimally augment the cholinergic system in the elderly and raise levels of the neurotransmitter, acetylcholine (ACh). Extensive studies involving chemistry, X-ray crystallography, biochemistry and pharmacology resulted in the design and synthesis of novel compounds to differentially inhibit either AChE or BChE in either the brain or periphery for an optimal duration for the potential treatment of a variety of diseases, such as AD, Myasthenia Gravis, and as chemical warfare prophylactics (collaborators: Drs. Brossi, Lahiri, Shafferman, Sambamurti, Descamp). In addition, specific and highly selective BChE inhibitors have been designed to characterize the role of this enzyme in brain during health, aging and disease. 1.1A. AChE: Two of our numerous novel synthesized AChE inhibitors are in development for the treatment of AD; specifically, the pure non-competitive inhibitors, phenserine and tolserine. Both are phenylcarbamates of physostigmine that are 70- and 190-fold selective for AChE vs. BChE. They have a favorable toxicologic profile and robustly enhance cognition in animal models (collaborator Dr. Ingram). They possess a long duration of reversible enzyme inhibition, coupled with a short pharmacokinetic half-life. This reduces dosing frequency, decreases body drug exposure and minimizes the dependence of drug action on the individual variations of drug metabolism commonly found in the elderly. In collaboration with industry, phenserine translated from the laboratory and into clinical trials where actions on cognition and levels of CSF and plasma Abeta have been assessed in mild to moderate AD (collaborators: Drs. Nordberg, Sambamurti, Lahiri). In parallel studies in collaboration with Dr. Utsuki (LSU), Dr. Irie (Kumomoto Univ., Japan) a transdermal patch has been developed to maintain steady-state optimal drug levels and maximize dosing compliance. 1.1B. BChE: In normal brain, some 80% of cholinesterase activity is in the form of AChE and 20% is BChE. AChE activity is concentrated mainly in neurons, while BChE is primarily associated with glial cells. Kinetic evidence indicates a role for BChE, in hydrolysing excess ACh. In advanced AD, however, AChE activity decreases to 15% of normal levels in affected brain regions, whereas BChE activity increases. The normal ratio of BChE to AChE becomes mismatched in AD causing excess metabolism of already depleted levels of ACh. The first available reversible and highly potent BChE inhibitors have been synthesized and are in preclinical assessment to evaluate their potential as AD drug candidates. On going studies are focusing on cognition and the molecular mechanisms underpinning AD with a focus to advance a BChE inhibitor to clinic assessment. The selective BChE inhibitor, (-)-bisnorcymserine, has been chosen and is advancing through required preclinical studies. Additional studies are utilizing these valuable agents to define the role of BChE in brain in health, aging and disease (collaborators: Drs. Klein, Lahiri) 1.2. Molecular events associated with AD: The reduction in levels of the potentially toxic amyloid-beta peptide (Abeta) has emerged as an important therapeutic goal in AD. Key targets for this goal are factors that affect the expression and processing of the Abeta precursor protein (APP). Our studies show that phenserine, reduces APP and Abeta levels in vivo and in tissue culture without toxicity. This activity is independent of its cholinesterase action, but is post-transcriptional: lowering APP protein levels without affecting mRNA levels. This is mediated in part via the 5-untranslated region (UTR) of APP mRNA. Current studies are characterizing mechanisms involved and focusing on these in the design and synthesis of agents that lower APP levels as a way of lower Abeta peptide (collaborators: Drs. Lahiri, Sambamurti, Rogers, Giordano, Utsuki). The compound, posiphen, has advanced to clinical trials and backup compounds are being assessed to undertsand molecular mechanisms underpinning activity. A series of new compounds has been designed that combine actions on Abeta with those on AChE or BChE at the same optimal concentration 2. Stroke, Parkinsons disease (PD), brain trauma: Drugs currently used provide temporary relief of symptoms, but do not prevent the cell death. Our target for drug design is the transcription factor, p53. Its up-regulation is a common feature of several neurodegenerative disorders, and is a gate keeper to the biochemical cascade that leads to apoptosis (programmed cell death). We recently designed and synthesized a novel series of tetrahydrobenzothiazole and oxazole analogues that inhibit p53 activity. Compounds are in current assessment for neuroprotective action in tissue culture and animal models (collaborators: Drs. Mattson, Ovadia, Pick, Hoffer, Wang) to select agents of potential for evaluation as drug candidates. Compounds of this calss have demonstrated biological activity in cellular andor animal models of stroke, AD and PD, thet are being assessed in other neurodegenerative diseases to define their optimal use. 3. Diabetes: Type 2 diabetes is a prevalent disease in the elderly. Present treatments are unsatisfactory. Our target for drug design is the glucagon-like peptide-1 (GLP-1) receptor (R). GLP-1 is secreted from the gut in response to food and is a potent secretagogue it binds to the GLP-1R on pancreatic beta-cells to induce glucose-dependent insulin secretion, thereby controling plasma glucose levels. We are developing long-acting GLP-1 analogues (collaborators: Drs. Egan, Mattson). This research aided in the development of the peptide exendin-4 (Ex-4) into clinical studies in type 2 diabetes. Novel chimeric peptides that combine the best features of GLP-1 and Ex-4 have also been designed and are under preclinical assessment in a variety models. We are characterizing the role of the GLP-1R stimulation in the nervous system, as it is found present in brain and peripheral nerve. GLP-1 analogues possess neurotrophic properties and protect neuronal cells from oxidative and Abeta-induced cell death. Neuroprotection in cell culture translated to in vivo studies in classical rodent neurodegeneration models, which include AD and peripheral neuropathy. Current studies are focused on selecting agents for clinical assessment. 4. Inflammation: Inflammation is a critical feature of neurodegereation and also occurs in numerous systemic diseases. Our target is the cytokine, TNF-alpha. Novel, potent TNF-alpha inhibitors are being synthesized on the backbone of thalidomide. They reduce TNF-alpha synthesis post-transcriptionally, via its 3-UTR, in cell culture studies. These are being assessed in vivo to define time- and concentration inhibition of TNF-alpa systemically as well as in brain. Upto 90% inhibition can be achieved in either compartment. Classical animals models are be utilized to aid in the selection of a clinical cadidate for chronic diseases such as amyotrophic lateral sclerosis and PD, as well as acute events such as head trauma (collaborators: Drs. Pick, Hoffer, Wang, Utsuki, Ingram Gabbita).

1.阿尔茨海默病：正在开发三个系列的药物来治疗阿尔茨海默病。乙酰胆碱酯酶 (AChE)、丁酰胆碱酯酶 (BChE) 和淀粉样β肽 (Abeta) 生成的选择性抑制剂。 1.1.胆碱酯酶抑制剂：开发的化合物可以最佳地增强老年人的胆碱能系统并提高神经递质乙酰胆碱 (ACh) 的水平。涉及化学、X 射线晶体学、生物化学和药理学的广泛研究导致设计和合成了新型化合物，以差异性地抑制大脑或外周中的 AChE 或 BChE，以达到最佳持续时间，从而可能治疗 AD、重症肌无力等多种疾病，并作为化学战预防剂（合作者：Brossi 博士、Lahiri 博士、 谢弗曼、桑巴穆尔蒂、德坎）。此外，还设计了特异性和高度选择性的 BChE 抑制剂来表征这种酶在健康、衰老和疾病期间在大脑中的作用。 1.1A。 AChE：我们正在开发众多新型合成的 AChE 抑制剂中的两种，用于治疗 AD；具体来说，是纯非竞争性抑制剂苯酚酚和托丝氨酸。两者都是毒扁豆碱的苯基氨基甲酸酯，对 AChE 的选择性是 BChE 的 70 倍和 190 倍。它们具有良好的毒理学特征，并能显着增强动物模型的认知能力（合作者 Ingram 博士）。它们具有长持续时间的可逆酶抑制作用，并且药代动力学半衰期短。这减少了给药频率，减少了体内药物暴露，并最大限度地减少了药物作用对老年人常见的药物代谢个体差异的依赖性。与工业界合作，苯酚从实验室转化为临床试验，在轻度至中度 AD 中评估了其对认知以及 CSF 和血浆 Abeta 水平的作用（合作​​者：Nordberg、Sambamurti、Lahiri 博士）。在与 Utsuki 博士（路易斯安那州立大学）、Irie 博士（日本熊本大学）合作的平行研究中，开发了一种透皮贴剂，以维持稳态最佳药物水平并最大限度地提高给药依从性。 1.1B。 BChE：在正常大脑中，约 80% 的胆碱酯酶活性以 AChE 形式存在，20% 为 BChE。 AChE 活性主要集中在神经元中，而 BChE 主要与神经胶质细胞相关。动力学证据表明 BChE 在水解过量的 ACh 中发挥作用。然而，在晚期 AD 中，受影响大脑区域的 AChE 活性降低至正常水平的 15%，而 BChE 活性则增加。 AD 中 BChE 与 AChE 的正常比例变得不匹配，导致已经耗尽的 ACh 水平过度代谢。第一个可用的可逆且高效的 BChE 抑制剂已经合成，并正在进行临床前评估，以评估其作为 AD 候选药物的潜力。正在进行的研究重点是认知和 AD 的分子机制，重点是将 BChE 抑制剂推进临床评估。选择性 BChE 抑制剂 (-)-双去甲丝氨酸已被选中，并正在通过所需的临床前研究进行进展。其他研究正在利用这些有价值的药物来确定 BChE 在大脑健康、衰老和疾病中的作用（合作​​者：Klein 博士、Lahiri 博士） 1.2.与 AD 相关的分子事件：降低潜在毒性的淀粉样β肽 (Abeta) 水平已成为 AD 的重要治疗目标。该目标的关键目标是影响 Abeta 前体蛋白 (APP) 表达和加工的因素。我们的研究表明，苯酚酚可降低体内和组织培养中 APP 和 Abeta 的水平，且无毒性。该活性与其胆碱酯酶作用无关，但属于转录后活性：降低 APP 蛋白水平而不影响 mRNA 水平。这部分是通过 APP mRNA 的 5 个非翻译区 (UTR) 介导的。目前的研究正在表征所涉及的机制，并重点关注降低 APP 水平的药物的设计和合成，作为降低 Abeta 肽的一种方式（合作者：Lahiri、Sambamurti、Rogers、Giordano、Utsuki 博士）。该化合物 Posiphen 已进入临床试验阶段，并且正在评估备用化合物，以了解支持活性的分子机制。设计了一系列新化合物，在相同的最佳浓度下将对 Abeta 的作用与对 AChE 或 BChE 的作用结合起来 2.中风、帕金森病(PD)、脑外伤：目前使用的药物可以暂时缓解症状，但不能阻止细胞死亡。我们的药物设计目标是转录因子 p53。它的上调是几种神经退行性疾病的共同特征，并且是导致细胞凋亡（程序性细胞死亡）的生化级联的看门人。我们最近设计并合成了一系列新型四氢苯并噻唑和恶唑类似物，可抑制 p53 活性。目前正在组织培养和动物模型中评估化合物的神经保护作用（合作​​者：Mattson、Ovadia、Pick、Hoffer、Wang 博士），以选择有潜力作为候选药物进行评估的药物。此类化合物已在中风、AD 和 PD 的细胞和/或动物模型中表现出生物活性，并且正在其他神经退行性疾病中对其进行评估，以确定其最佳用途。 3、糖尿病：2型糖尿病是老年人的多发病。目前的治疗效果并不令人满意。我们的药物设计目标是胰高血糖素样肽-1 (GLP-1) 受体 (R)。 GLP-1 响应食物而从肠道分泌，是一种有效的促分泌剂，它与胰腺 β 细胞上的 GLP-1R 结合，诱导葡萄糖依赖性胰岛素分泌，从而控制血浆葡萄糖水平。我们正在开发长效 GLP-1 类似物（合作者：Egan 博士、Mattson 博士）。 这项研究有助于将肽 exendin-4 (Ex-4) 开发到 2 型糖尿病的临床研究中。结合了 GLP-1 和 Ex-4 最佳特征的新型嵌合肽也已被设计出来，并正在各种模型中进行临床前评估。我们正在描述 GLP-1R 刺激在神经系统中的作用，因为它存在于大脑和周围神经中。 GLP-1 类似物具有神经营养特性，可保护神经元细胞免受氧化和 Abeta 诱导的细胞死亡。细胞培养中的神经保护转化为经典啮齿动物神经变性模型（包括 AD 和周围神经病变）的体内研究。目前的研究重点是选择用于临床评估的药物。 4.炎症：炎症是神经退行性疾病的一个重要特征，也发生在许多全身性疾病中。我们的目标是细胞因子 TNF-α。新型、有效的 TNF-α 抑制剂正在沙利度胺的基础上合成。在细胞培养研究中，它们通过 3-UTR 减少转录后 TNF-α 的合成。这些正在体内进行评估，以确定全身和大脑中 TNF-α 的时间和浓度抑制。任一隔室均可实现高达 90% 的抑制。经典动物模型可用于帮助选择慢性疾病（如肌萎缩侧索硬化症和 PD）以及急性事件（如头部创伤）的临床候选者（合作者：Pick、Hoffer、Wang、Utsuki、Ingram Gabbita 博士）。

项目成果

Novel p53 inactivators with neuroprotective action: syntheses and pharmacological evaluation of 2-imino-2,3,4,5,6,7-hexahydrobenzothiazole and 2-imino-2,3,4,5,6,7-hexahydrobenzoxazole derivatives.

• DOI: 10.1021/jm020044d
• 发表时间: 2002-10
• 期刊: Journal of medicinal chemistry
• 影响因子: 7.3
• 作者: XiaoXiang Zhu;Qian-sheng Yu;R. Cutler;C. Culmsee;H. Holloway;D. Lahiri;M. Mattson;N. Greig

Syntheses of tetrahydrofurobenzofurans and dihydromethanobenzodioxepines from 5-hydroxy-3-methyl-3H-benzofuran-2-one. Rearrangement and ring expansion under reductive conditions on treatment with hydrides.

由 5-羟基-3-甲基-3H-苯并呋喃-2-酮合成四氢呋喃苯并呋喃和二氢甲烷苯并二氧杂环己烷。

• DOI: 10.1021/jo0503052
• 发表时间: 2005
• 期刊: The Journal of organic chemistry
• 作者: Luo,Weiming;Yu,Qian-Sheng;Holloway,HaroldW;Parrish,Damon;Greig,NigelH;Brossi,Arnold
• 通讯作者: Brossi,Arnold

Characterization of the human beta-secretase 2 (BACE2) 5'-flanking region: identification of a 268-bp region as the basal BACE2 promoter.

人 β 分泌酶 2 (BACE2) 5 侧翼区域的表征：鉴定 268 bp 区域作为基础 BACE2 启动子。

• DOI: 10.1385/jmn:29:1:81
• 发表时间: 2006
• 期刊: Journal of molecular neuroscience : MN
• 作者: Maloney,Bryan;Ge,Yuan-Wen;Greig,NigelH;Lahiri,DebomoyK
• 通讯作者: Lahiri,DebomoyK

Tomographic visualization of cholinesterase.

胆碱酯酶的断层扫描可视化。

• DOI: 10.1002/ana.20926
• 发表时间: 2006
• 期刊: Annals of neurology
• 影响因子: 11.2
• 作者: Kuljis,RodrigoO;Darvesh,Sultan;Greig,NigelH;Geula,Changiz
• 通讯作者: Geula,Changiz

Does nitric oxide synthase contribute to the pathogenesis of Alzheimer's disease?: effects of beta-amyloid deposition on NOS in transgenic mouse brain with AD pathology.

一氧化氮合酶是否与阿尔茨海默氏病的发病机制有关？：β-淀粉样蛋白沉积对患有 AD 病理的转基因小鼠大脑中 NOS 的影响。

• DOI: 10.1196/annals.1299.117
• 发表时间: 2003
• 期刊: Annals of the New York Academy of Sciences
• 影响因子: 5.2
• 作者: Lahiri,DK;Chen,D;Ge,Y-W;Farlow,M;Kotwal,G;Kanthasamy,A;Ingram,DK;Greig,NH
• 通讯作者: Greig,NH