Development of GSK-3 Beta Inhibitors for the Treatment of Parkinson's Disease

开发用于治疗帕金森病的 GSK-3 Beta 抑制剂

• 批准号: 7596052
• 负责人: Jia Zhou
• 金额: $ 28.26万
• 依托单位: PSYCHOGENICS, INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7596052
• 关键词: 1-Methyl-4-phenylpyridinium; Adverse effects; Affect; Age; Aging; Agreement; Alzheimer' s Disease; Animals; Arizona; Arts; Autopsy; Benzofurans; Biochemistry; Bipolar Disorder; Brain; Brain region; Catechols; Cells; Central Nervous System Diseases; Chemistry; Chicago; Chronic; Collaborations; Comorbidity; Complex; Condition; Cytoskeleton; Data; Degenerative Disorder; Development; Disease; Dissociation; Dopamine Agonists; Dose; Ensure; Equipment; Family; Frontotemporal Dementia; Funding; Genes; Glycogen Synthase Kinase 3; Glycogen Synthase Kinases; Goals; Grant; Hand; Housing; Human; Illinois; In Vitro; Inclusion Bodies; Indazoles; Indoles; Injection of therapeutic agent; Investigation; Laboratory Research; Lead; Legal patent; Lesion; Levodopa; Lewy Bodies; Libraries; Licensing; Ligands; Link; Lithium; Maleimides; Modeling; Modification; Molecular Biology; Mus; Mutation; Nerve; Nerve Degeneration; Neurodegenerative Disorders; Neurofibrillary Tangles; Neurologic; Neurons; Neuroprotective Agents; Neurosciences; Numbers; Organic Chemistry; Parkinson Disease; Parkinson' s Dementia; Parkinsonian Disorders; Pathology; Patients; Pattern; Personal Satisfaction; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacotherapy; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Proteins; Public Health; Relative (related person); Research; Research Proposals; Role; Science; Screening procedure; Senior Scientist; Single Nucleotide Polymorphism; Site; Small Business Technology Transfer Research; Societies; Specific qualifier value; Staurosporine; Symptoms; Tauopathies; Technology; Testing; Therapeutic; Time; Toxin; Transferase; Treatment Efficacy; Triton X100; Universities; Work; age related; analog; base; benzofuran; chemical synthesis; cytotoxicity; day; design; drug development; experience; follow-up; hyperphosphorylated tau; in vivo; indole; inhibitor/antagonist; kinase inhibitor; neuroprotection; neurorestoration; novel; payment; presynaptic; prevent; response; sarkosyl; scale up; small molecule; synuclein; synucleinopathy; tau Proteins; tau aggregation; tau-protein kinase

DESCRIPTION (provided by applicant): Parkinson's disease (PD), a chronic and progressive neurological condition and one of aging and age- related diseases, affects approximately 1.5 million people in the US alone. PD not only places severe burden on the patients but also on their family and society. In 2003, approximately US $2.3 billion was spent on drug therapy worldwide to treat Parkinson's disease. Although great strides have been made in the development of agents to treat PD, the existing PD drugs such as dopamine (DA) agonists, levodopa and catechol-O-methyl transferase inhibitors (COMT) only treat the symptoms of the disease, and are also fraught with adverse effects and long-term complications. Consequently, there is a great need for developing disease-modifying and neuroprotective as well as neurorestorative drugs which can slow down or stop the disease from progressing. Recent research suggests that inhibition of the glycogen synthase kinase-3¿ (GSK-3¿) by small molecules may offer an important strategy in the treatment of a number of neurodegenerative diseases including Alzheimer's disease (AD), but its usefulness in PD has not been described. We have recently found in the cellular and animal MPTP model of parkinsonism, an induction in the hyperphosphorylated form of Tau, p-Tau, with hyperphosphorylation seen at many sites, including those found in neurofibrillary tangles of AD. The increase in p-Tau levels was strictly dependent upon the presence of a- Syn. This requirement for a-Syn was mandatory, since in a-Syn-/- mice, and in transfected cells not expressing any a-Syn, the toxin failed to induce p-Tau. MPTP also caused an increase in a-Syn protein levels. Hyperphosphorylation of Tau lead to its dissociation from the cytoskeleton, and aggregates of p-Tau were seen in sarkosyl-insoluble and Triton X-100-insoluble fractions. a-Syn was able to form stable heteromeric protein complexes with p-Tau, and p-Tau aggregates were seen in mature inclusion bodies of a-Syn. MPP+ caused the activation of several p-Tau-specific kinases, such as GSK-3¿ and p-ERK. Blockade of GSK-3¿ not only prevented, but also reduced, MPP+-induced p-Tau formation, a-Syn accumulation and cytotoxicity. Very new data obtained in human postmortem PD brains, in collaboration with Dr. Jeffrey Joyce, show a similar pattern of pathology: increased a-Syn accumulation, hyperphosphorylation of Tau at sites similar to the MPTP models [pSer262 and pSer396/404], lack of phosphorylation at sites not seen with MPTP [pSer202], and large increases in GSK-3¿. Interestingly, these pathological changes were further augmented in PD patients with dementia [PD + DEM]. Together, these findings in PD brains confirm the validity of our findings with the MPTP models. Importantly, prior to our findings, a possible role for GSK-3¿ in PD has not been previously described, although its role in AD is well studied. In addition to our findings, another study found strong linkage of two single nucleotide polymorphisms in the GSK-3¿ gene to sporadic PD. Thus, GSK-3¿ presents a novel target site in the development of novel therapies for PD. To date, we have identified some nM potency GSK-3¿ inhibitors that emerged from our SAR studies of staurosporine. A number of these designed staurosporine analogs have been screened against a family of 30 kinases. Among the compounds tested we found one, an indolyl-indazolylmaleimide, that was able to inhibit 98% of the kinase activity of GSK-3¿ when tested at a concentration of 10 ¿M. After further structural modifications described below, novel 3-(indol-3-yl)-4-(benzofuran-3-yl)maleimides having a Ki value as low as 2 nM selectively against GSK-3¿ relative to 30 additional kinases were identified. Moreover, we have been able to show that some of these ligands are able to exert a neuroprotective and neurorestorative action in vitro. The ultimate goal would be to identify one or two GSK-3¿ inhibitors that could be further developed for slowing or halting the progression of PD. Through funding from this STTR grant, we intend to follow-up on the exciting preliminary findings we have made in pursuit of novel GSK-3¿ inhibitors as potential therapeutics for the treatment of PD. To achieve this goal, the Specific Aims of this research proposal are as follows: 1. Compound selection and synthesis: Based upon the compound library in hand, 10 potent GSK- 3¿ inhibitors will be resynthesized for further in vitro pharmacological studies. Then 2 or 3 of the most promising ligands based upon the in vitro profile will be scaled up (about 5 grams each) for in vivo animal studies. 2. Neuroprotective In vitro studies in transfected cells and neurons: Investigation of time course and dose response of the compounds for inhibition of GSK-3¿, analyses of other kinases and phosphatases that are modulated by these compounds, and comparison of our findings with the effects using lithium. 3. Neurorestorative in vitro studies in transfected cells and neurons: After the initiation of cytotoxicity, we will examine the time and dose of GSK-3¿ inhibitors necessary for neurorestoration. 4. In vivo animal studies: Injection with MPTP for 5 days will be followed by simultaneous or delayed injections with increasing doses of the GSK-3¿ inhibitors for different time periods. GSK-3¿, a-Syn, Tau, kinases and phosphatases will be examined, as specified in Specific Aims 2 and 3. Key words: GSK-3¿ inhibitors, Kinases, Selectivity, Parkinson's disease, Therapeutics, Staurosporine analogs, Neuroprotective agents, Neurorestorative agents, Neuroprotection, a-Synuclein, p-Tau, Taupathies, Synucleopathies, MPTP model, Aging, Neurodegenerative, Age-related diseases, Alzheimer's disease. Project Des inson's disease (PD), the second most common neurodegenerative disease next to Alzheimer's disease (AD), is a progressive neurological condition associated with aging. Our proposal entails the chemical synthesis of novel staurosporine analogues as potent glycogen synthase kinase-3¿ (GSK-3¿) inhibitors, and in vitro studies in transfected cells and neurons as well as in vivo studies in animals of these compounds, which have the great potential to be developed as neuroprotective and neurorestorative therapies to slow down or stop Parkinson's disease from progressing. Public Health

描述（由适用提供）：帕金森氏病（PD），一种慢性和进行性神经系统疾病，一种与年龄和年龄有关的疾病之一，仅在美国就会影响约150万人。 PD不仅对患者造成严重烧伤，而且对他们的家庭和社会施加了严重的烧伤。 2003年，全世界在药物治疗上花费了约23亿美元来治疗帕金森氏病。尽管在治疗PD的药物的开发中取得了长足的进步，但现有的PD药物（例如多巴胺（DA）激动剂，左旋多巴和Catechol-O-O-甲基转移抑制剂（COMT）仅治疗疾病的症状，并且也具有不良影响和长期的并发症。因此，非常需要发展疾病改良和神经保护性以及神经抑制药物，这些药物可以减慢或阻止疾病的发展。最近的研究表明，小分子抑制糖原合成酶-3？（GSK-3？）可能在治疗许多神经退行性疾病（包括阿尔茨海默氏病（AD））方面提供了重要的策略，但尚未描述其在PD中的有用性。我们最近在帕金森氏症的细胞和动物MPTP模型中发现了tau，p-tau的热磷酸化形式的诱导，在许多部位都可以看到过度磷酸化，其中包括在AD的神经纤维缠结中发现的。 P-TAU水平的增加严格取决于A-Syn的存在。对于A-Syn的要求是强制性的，因为在A-Syn - / - 小鼠中，在不表达任何A-Syn的翻译细胞中，毒素未能诱导P-TAU。 MPTP还导致A-SYN蛋白水平升高。 tau的高磷酸化导致其与细胞骨架的解离，并且在sarkosyl-syl-syl-syl-by-bion-n-Sarkosyl-syl-n-bears-bys-bysy和triton X-100不可溶的分数中看到了P-TAU的聚集体。 A-Syn能够与P-TAU形成稳定的异源蛋白复合物，并在A-Syn的成熟包含体中看到P-TAU聚集体。 MPP+引起了几种P-TAU特异性激酶的激活，例如GSK-3和P-ERK。 GSK-3的封锁不仅防止了，还可以减少MPP+诱导的P-TAU形成，A-SYN积累和细胞毒性。与杰弗里·乔伊斯（Jeffrey Joyce）博士合作，在人类后PD大脑中获得的非常新的数据显示了类似的病理模式：增加A-Syn积累，与MPTP模型[PSER262和PSER396/404]相似的站点的热磷酸化，缺乏MPTP [PSER396/404]，缺乏磷酸化的位点[and 3]和PSERS222的磷酸化。有趣的是，PD患者[PD + DEM]的PD患者进一步增加了这些病理变化。总之，PD大脑中的这些发现证实了我们通过MPTP模型发现的有效性。重要的是，在我们的发现之前，尽管对其在AD中的作用进行了充分的研究，但尚未描述GSK-3`在PD中的可能作用。除了我们的发现外，另一项研究还发现，GSK-3域中两个单个核苷酸多态性与零星PD的紧密联系。这是GSK-3“在开发PD的新疗法中提出了一个新的目标部位。迄今为止，我们已经确定了一些NM效力GSK-3“抑制剂，这些抑制剂来自我们的星形孢菌素研究。这些设计的星形孢菌素类似物中的许多已针对30个激酶的家族进行了筛选。 Among the compounds tested we found one, an indolyl-indazolylmaleimide, that was able to inhibit 98% of the kinase activity of GSK-3¿ When tested at a concentration of 10 ¿ M. After further structural modifications described below, novel 3-(indol-3-yl)-4-(benzofuran-3-yl)maleimides having a Ki value as low as 2 nM selectively against GSK-3¿相对于另外30种激酶。此外，我们已经能够证明其中一些配体能够在体外执行神经保护作用和神经养生作用。最终目标是识别一种或两个GSK-3“抑制剂，这些抑制剂可以进一步开发，以减慢或停止PD的进展。通过这笔STTR赠款的资助，我们打算跟进我们为追求新颖的GSK-3“抑制剂作为PD治疗的潜在治疗剂，我们已经提出的令人兴奋的初步发现。为了实现这一目标，该研究建议的具体目的如下：1。化合物选择和合成：基于手头的复合库，将重新合成10个潜在的GSK-3“抑制剂，以进一步进行体外药理研究。然后，将根据体外谱的2或3个最有前途的配体进行体内动物研究的缩放（每个约5克）。 2。转染细胞和神经元中的神经保护性研究：对抑制GSK-3？的时间过程和剂量反应的研究，对这些化合物调节的其他激酶和磷酸酶的分析，以及对我们的发现与使用锂的作用进行比较的分析。 3。转染细胞和神经元中的神经培养性在体外研究：在细胞毒性开始后，我们将检查GSK-3“ GSK-3”抑制剂的时间和剂量。 4。体内动物研究：用MPTP注射5天之后，随后是简单或延迟的注射，在不同时间段的GSK-3？抑制剂增加。将检查GSK-3，A-SYN，TAU，激酶和磷酸酶，如特定目的2和3所示。 taupathies，nulepathies，MPTP模型，衰老，神经退行性，与年龄相关的疾病，阿尔茨海默氏病。 Des Inson疾病（PD）是第二个最常见的神经退行性疾病（AD），是与衰老相关的进行性神经系统疾病。我们的提议需要化学合成新型Staurosporine类似物作为潜在的糖原合成酶激酶-3？（GSK-3¿）抑制剂，以及在转染细胞和神经元中进行的体外研究以及这些化合物的动物的体内研究，这具有较大的潜力，这些潜在的潜力却可以降低神经植物的过度培训或神经性的介绍性，从而逐渐脱离了过度培训。公共卫生