Novel Hsp90 Inhibitors: Proof-of-Principle for Treatment of Motor Neuron Diseases

新型 Hsp90 抑制剂：治疗运动神经元疾病的原理验证

• 批准号: 8054783
• 负责人: RUIHONG CHEN
• 金额: $ 14.23万
• 依托单位: NEXGENIX PHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8054783
• 关键词: Adopted; Adult; Age; Amyotrophic Lateral Sclerosis; Animal Model; Binding; Blood - brain barrier anatomy; Brain; Cells; Cessation of life; Client; Clinical; Complex; Cuprozinc Superoxide Dismutase; Data; Development; Diagnosis; Disease; Disease Progression; Dose; Drug Kinetics; Effectiveness; Embryo; Epigenetic Process; Experimental Models; Familial Amyotrophic Lateral Sclerosis; Foundations; Funding; Funding Agency; Funding Mechanisms; Gene Mutation; Genes; Genetic; Goals; HSP 90 inhibition; Heat shock proteins; Heat-Shock Proteins 90; In Vitro; Incidence; Inherited; Intervention; Lead; Link; Medical; Mission; Modeling; Modification; Molecular Chaperones; Molecular Conformation; Motor; Motor Neuron Disease; Motor Neurons; Mus; Mutation; National Institute of Neurological Disorders and Stroke; Neurologic; Neuromuscular Diseases; Pathway interactions; Penetration; Pharmaceutical Preparations; Pharmacodynamics; Play; Population; Post-Translational Protein Processing; Preclinical Testing; Process; Proteins; Quality Control; Receptor Signaling; Regimen; Research; Respiratory Insufficiency; Riluzole; Rodent; Role; Series; Signaling Molecule; Spinal Cord; Spinal Ganglia; System; Testing; Therapeutic; Tissues; Toxic effect; Transgenic Mice; Translational Research; Ubiquitin; United States National Institutes of Health; Up-Regulation; Work; base; dosage; drug development; heat shock transcription factor; in vivo; inhibitor/antagonist; intraperitoneal; mouse model; multicatalytic endopeptidase complex; mutant; novel; novel therapeutics; pre-clinical; prevent; programs; protein degradation; protein misfolding; public health relevance; research study; therapeutic development; tissue culture; transcription factor

DESCRIPTION (provided by applicant): Amyotrophic Lateral Sclerosis (ALS or Lou Gherig's disease) is an adult onset motor neuron disease characterized by progressive loss of motor function and death from respiratory insufficiency, usually within five years of diagnosis. No adequate treatment exists. Most cases of ALS are sporadic; however, several genetic loci have been linked to familial forms of the disease, the most common (fALS1) being dominantly inherited mutations in the gene encoding Cu/Zn-superoxide dismutase (SOD1). Experimental models of fALS1 are used in preclinical testing of compounds for eventual use in treating sporadic as well as familial ALS. Given that ALS can be caused by multiple, mostly unidentified insults, common mechanisms of disease progression and general neuroprotective pathways are targets for intervention. Objective/Hypothesis: The long term objective is to develop a therapy for ALS and other motor neuron disorders based on a novel series of compounds that inhibit Hsp90, a heat shock protein (HSP) that plays a pivotal role in regulating folding and turnover of proteins with aberrant conformation. The underlying hypothesis is that these compounds will be efficacious through two mechanisms that contribute to protein quality control: (1) upregulating multiple heat shock proteins (HSPs) with protein chaperoning and refolding function, thereby preventing damaged proteins (arising from genetic mutation or epigenetic modification) from adopting toxic conformations and aggregating, and (2) promoting degradation of Hsp90 client proteins, including mutant and misfolded proteins, thereby reducing the toxic burden. The short term objective is to evaluate our presumptive lead compound, NXD30001 in primary tissue culture models and a transgenic mouse model of fALS. The realization of our goals will help further the mission of the NINDS, to reduce the burden of neurologic disease. Specific Aim 1: To test NXD30001 in existing in vitro primary culture models of ALS. Test-models are established by (1) expressing ALS-causing mutant SOD1 proteins in motor neurons of dissociated cultures of embryonic murine spinal cord-dorsal root ganglia (DRG), and (2) preparing spinal cord-DRG cultures from mutant SOD1 transgenic mouse embryos. These experiments will establish bioactivity of NXD30001 in motor neurons, specifically expression of HSPs and increased turnover of Hsp90 client proteins, and will evaluate the effectiveness of NXD30001 in preventing toxicity of fALS-causing SOD1 mutants. Specific Aim 2. Perform proof-of-concept studies in an fALS animal model to determine the in vivo utility of the NXD30001 compound. The efficacy of NXD30001 will be assessed in transgenic mice expressing the disease-causing SOD1 mutant, SOD1G93A. Two dosage regimens will be evaluated for delaying onset and progression of motor neuron disease, using induction of HSPs and reduction in levels of of Hsp90 client proteins as markers of NXD30001 bioactivity in brain and spinal cord. PUBLIC HEALTH RELEVANCE: The only approved drug for treating Amyotrophic Lateral Sclerosis (ALS), riluzole, provides minimal benefit for this fatal disease. With an incidence of 1-2 per 100,000, ALS represents a serious unmet medical need, which will only grow as the population ages. The proposed research may provide proof-of-principle evidence for the inhibition of heat shock protein 90 (Hsp90) as a potential approach for treating ALS, and will likely accelerate the process of discovery and development of novel therapeutics for ALS and other motor neuron disorders.

描述（由申请人提供）：肌萎缩侧索硬化症（ALS或Lou Gherig病）是一种成人发病的运动神经元疾病，其特征是运动功能进行性丧失和呼吸功能不全死亡，通常在诊断后5年内发生。没有适当的治疗。大多数ALS病例是散发性的;然而，几个遗传基因座与该疾病的家族形式有关，最常见的（fALS1）是编码Cu/Zn-超氧化物歧化酶（SOD1）的基因中的显性遗传突变。fALS1的实验模型用于化合物的临床前测试，以最终用于治疗散发性以及家族性ALS。鉴于ALS可由多种、大多数未识别的损伤引起，疾病进展的常见机制和一般神经保护途径是干预的目标。目的/假设：长期目标是开发基于抑制Hsp90的一系列新化合物的ALS和其他运动神经元病症的疗法，Hsp90是一种热休克蛋白（HSP），其在调节具有异常构象的蛋白质的折叠和周转中起关键作用。基本假设是这些化合物将通过有助于蛋白质质量控制的两种机制有效：（1）上调具有蛋白质伴侣和重折叠功能的多种热休克蛋白（HSP），从而防止受损的蛋白质（由基因突变或表观遗传修饰引起）避免采用毒性构象和聚集，和（2）促进Hsp90客户蛋白的降解，包括突变和错误折叠的蛋白质，从而减少毒性负担。短期目标是在原代组织培养模型和fALS转基因小鼠模型中评价我们假定的先导化合物NXD 30001。我们的目标的实现将有助于进一步的使命的NINDS，以减少神经系统疾病的负担。具体目的1：在现有ALS体外原代培养模型中检测NXD 30001。通过（1）在胚胎鼠脊髓-背根神经节（DRG）的分离培养物的运动神经元中表达引起ALS的突变体SOD 1蛋白，和（2）从突变体SOD 1转基因小鼠胚胎制备脊髓-DRG培养物来建立测试模型。这些实验将确定NXD 30001在运动神经元中的生物活性，特别是HSP的表达和Hsp90客户蛋白的周转增加，并将评估NXD 30001在预防引起fALS的SOD1突变体毒性方面的有效性。具体目标2。在fALS动物模型中进行概念验证研究，以确定NXD 30001化合物的体内效用。将在表达致病性SOD1突变体SOD1G93A的转基因小鼠中评估NXD 30001的疗效。将使用HSP诱导和Hsp90客户蛋白水平降低作为脑和脊髓中NXD 30001生物活性的标志物，评价两种给药方案延迟运动神经元疾病发作和进展的效果。 公共卫生相关性：唯一批准用于治疗肌萎缩侧索硬化症（ALS）的药物利鲁唑对这种致命疾病的益处最小。ALS的发病率为每10万人中有1 - 2人，是一种严重未满足的医疗需求，随着人口老龄化，这种需求只会增加。这项研究可能为抑制热休克蛋白90（Hsp90）作为治疗ALS的潜在方法提供原理性证据，并可能加速ALS和其他运动神经元疾病的新疗法的发现和开发过程。