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。阿尔茨海默氏病：正在开发三种制剂来治疗AD。乙酰胆碱酯酶（ACHE），丁酰胆碱酯酶（BCHE）和淀粉样蛋白β肽（ABETA）产生的选择性抑制剂。 1.1。胆碱酯酶抑制剂：开发化合物是为了最佳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。 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）。此外，已经设计了特定和高度选择性的BCHE抑制剂，以表征该酶在健康，衰老和疾病中的大脑中的作用。 1.1a。 ACHE：我们众多新型合成的ACHE抑制剂中有两个正在开发AD治疗。具体而言，纯的非竞争性抑制剂，保透明碱和托尔赛因。两者都是Physostigmine的苯基钙化，对于ACHE与BCHE的选择性为70倍和190倍。它们具有良好的毒性特征，并且可以在动物模型中强大增强认知（Ingram博士）。它们具有长时间的可逆酶抑制作用，再加上短的药代动力学半衰期。这会降低给药频率，降低人体药物的暴露，并最大程度地减少药物作用对老年人常见的药物代谢的个体变异的依赖性。在与行业合作的情况下，Phenserine从实验室转化为临床试验，在轻度至中度的AD中，已经评估了对CSF和血浆ABETA认知和血浆Abeta水平的行动（合作者：Nordberg博士，Sambamurti，Lahiri，Lahiri）。在与Utsuki博士（LSU）合作的并行研究中，Irie博士（日本Kumomoto Univ。）已经开发了透皮斑块，以维持稳态的最佳药物水平并最大程度地提高剂量依从性。 1.1b。 BCHE：在正常的大脑中，约80％的胆碱酯酶活性为ACHE，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有关。动力学证据表明BCHE在水解过量ACH中的作用。然而，在晚期AD中，ACHE活性在受影响的大脑区域中降至正常水平的15％，而BCHE活性增加。 BCHE与ACHE的正常比率在AD中不匹配，导致ACH水平耗尽的过量代谢。首次可用的可逆且高度有效的BCHE抑制剂已合成，并且正在临床前评估中评估其作为AD药物候选者的潜力。进行研究的重点是认知和基于AD的分子机制，重点是将BCHE抑制剂推向临床评估。选择性BCHE抑制剂（ - ） - 双诺菌丝氨酸已被选择，并通过所需的临床前研究进行前进。其他研究正在利用这些有价值的药物来定义BCHE在健康，衰老和疾病中的作用（合作​​者：克莱恩博士，lahiri） 1.2。与AD相关的分子事件：潜在毒性淀粉样蛋白β肽（ABETA）水平的降低已成为AD中重要的治疗目标。该目标的主要目标是影响Abeta前体蛋白（APP）表达和处理的因素。我们的研究表明，Phenserine在体内和组织培养中降低了APP和ABETA水平，而无需毒性。该活性与其胆碱酯酶作用无关，但在转录后是：降低APP蛋白水平而不会影响mRNA水平。这是通过App mRNA的5个非翻译区（UTR）部分介导的。当前的研究表征了涉及的机制，并专注于这些机制在降低App水平的设计和合成中，这些试剂降低了App含量的较低肽的方式（合作者：Lahiri博士，Sambamurti博士，Rogers，Rogers，Giordano，utsuki）。该化合物Posiphen已升级为临床试验，并且正在评估备份化合物，以实现和分子机制。设计了一系列新化合物，将Abeta上的动作与ACHE或BCHE上的动作结合起来，以相同的最佳浓度 2。中风，帕金森病（PD），脑创伤：当前使用的药物可暂时缓解症状，但不能防止细胞死亡。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常见特征，并且是导致凋亡的生化级联反应的门将（程序性细胞死亡）。我们最近设计并合成了一系列新型的四氢苯甲酸苯唑和抑制p53活性的恶唑类似物。在组织培养和动物模型中的神经保护作用的当前评估中（合作者：马特森博士，奥瓦迪亚博士，挑选，霍夫，王，王），以选择作为候选药物评估的潜力的药物。这些CALS的化合物在其他神经退行性疾病中评估了中风，AD和PD的细胞Andor动物模型，以定义其最佳用途。 3.糖尿病：2型糖尿病是老年人普遍存在的疾病。目前的治疗不令人满意。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是从肠道中分泌的，是对食物的响应，是一种有效的促分泌物，它与胰腺β细胞上的GLP-1R结合，以诱导葡萄糖依赖性胰岛素分泌，从而控制血浆葡萄糖水平。我们正在开发长效的GLP-1类似物（合作者：Egan博士，马特森）。 这项研究有助于将肽Exendin-4（EX-4）开发到2型糖尿病中的临床研究中。结合GLP-1和EX-4最佳特征的新型嵌合肽也已设计并在各种模型中的临床前评估。我们正在表征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的特性，并保护神经元细胞免受氧化和ABETA诱导的细胞死亡的影响。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的体内研究，其中包括AD和周围神经病。当前的研究集中在选择临床评估的药物上。 4。炎症：炎症是神经残疾人的关键特征，也发生在许多全身性疾病中。我们的靶标是细胞因子TNF-α。新颖的有效TNF-α抑制剂正在甲状酸酯的支柱上合成。它们在细胞培养研究中通过其3-UTR在转录后降低TNF-α合成。这些正在体内评估，以便在系统和大脑中定义TNF-ALPA的时间和浓度抑制。在任何一个隔间中，最多可抑制90％。使用古典动物模型来帮助选择慢性疾病（如肌萎缩性侧面硬化症和PD）的临床减少症，以及诸如头部创伤之类的急性事件（合作者：Pick。Pick，Hoffer，Wang，Wang，Utsuki，Utsuki，Ingram Gabbita）。

项目成果

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

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

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

Editorial: advances in Alzheimer therapy: something old, something new, something borrowed, something blue.

社论：阿尔茨海默病治疗的进展：旧的、新的、借来的、蓝色的。

• DOI: 10.2174/1567205054367865
• 发表时间: 2005
• 期刊: Current Alzheimer research
• 影响因子: 2.1
• 作者: Greig,NigelH;Lahiri,DebomoyK;Giacobini,Ezio
• 通讯作者: Giacobini,Ezio