Optimization of HDAC6 Inhibitors in the Treatment of CMT

HDAC6 抑制剂治疗 CMT 的优化

• 批准号: 8814283
• 负责人: Ludo Van Den Bosch
• 金额: $ 37.77万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8814283
• 关键词: Acetylation; Adolescence; Adult; Affect; Age-Months; Animal Model; Animals; Axon; Axonal Transport; Behavior; Cells; Characteristics; Charcot-Marie-Tooth Disease; Chemicals; Claw; Clinic; Computer Simulation; Deacetylation; Defect; Deformity; Disease; Distal; Ensure; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Family; Family member; Foot-drop; Gait; Genes; Grant; Guidelines; HDAC6 gene; Hammer Toes; Hand; Health; Heat Shock Protein 27; Heat shock proteins; Hereditary Motor and Sensory Neuropathies; Histone Deacetylase; Histone Deacetylase Inhibitor; In Vitro; Individual; Inherited; Inhibitory Concentration 50; Leg; Legitimacy; Limb structure; Measurement; Methods; Mitochondria; Monitor; Motor; Movement; Mus; Muscle; Muscle Weakness; Muscular Atrophy; Mutant Strains Mice; Mutation; Names; Neuromuscular Diseases; Neurons; Neuropathy; Outcome; Paris, France; Patients; Pattern; Performance; Peripheral; Peripheral Nerves; Permeability; Pharmaceutical Chemistry; Pharmaceutical Preparations; Physicians; Protein Isoforms; Research Personnel; Role; Rotarod Performance Test; Safety; Sensory; Severities; Spinal Ganglia; Staging; Structure-Activity Relationship; Symptoms; Tail; Tail Suspension; Talipes cavus; Testing; Therapeutic; Tooth structure; Transgenic Animals; Transgenic Mice; Tubulin; United States National Institutes of Health; Work; afferent nerve; analog; arm; axonopathy; design; disease phenotype; emerging adult; falls; foot; improved; inhibitor/antagonist; mouse model; mutant; nerve supply; nervous system disorder; programs

DESCRIPTION (provided by applicant): Charcot-Marie-Tooth disease (CMT) is one of the most common inherited neurological disorders that affects ~ 1 in 2,500 people in the US. The disease is named for the three physicians who first identified it in 1886. CMT, also known as hereditary motor and sensory neuropathy (HMSN) or peroneal muscular atrophy, is made up of a group disorders that affect peripheral nerves. CMT affects both motor and sensory nerves. A typical characteristic includes weakness of the foot and lower leg muscles, which may result in foot drop and a high-stepped gait with frequent tripping or falls. Mutations in the small heat-shock protein 27 (HSPB1) cause axonal CMT or distal hereditary motor neuropathy (distal HMN). Our team has developed and characterized transgenic mice expressing mutant (S135F) HSPB1 in neurons only. These mice show all features of CMT2. Expression of mutant HSPB1 decreased acetylated ¿-tubulin levels and induced severe axonal transport deficits. Of great significance, we have now shown that pharmacological inhibition of histone deacetylase 6 (HDAC6)-induced ¿-tubulin deacetylation using our designed HDAC6I [HDAC6 Inhibitor] Tubastatin A corrected the axonal transport defects induced by HSPB1 mutations and rescued the CMT phenotype of symptomatic mutant HSPB1 mice. Our findings demonstrate the pathogenic role of ¿-tubulin deacetylation in mutant HSPB1-induced neuropathies and offer valuable perspectives for HDAC6 inhibitors as a therapeutic strategy for hereditary axonopathies. In order to extend these high impact findings and to advance the best HDAC6I therapies to the clinic for the treatment of CMT and related neuropathies, we intend to more fully explore the structure activity relationships [SAR] of a family of Tubastatin A analogs, the key HDAC inhibitor used in our studies. The aims of this proposal are as follow: 1. Using the principles of medicinal chemistry and computer modeling methods, design and synthesize improved analogs of Tubastatin A and further optimize these for their in vitro HDAC isoform selectivity and potency. In this aim, in silico ADMET calculations will be performed so that only the most drug-like compounds are prepared for the preliminary in vitro studies. 2. For those compounds showing at least 300-fold selectivity for HDAC6 over the other isoforms and having an IC50 of < 50 nM, optimize these new chemical entities for their ability to enhance tubulin acetylation and mitochondrial movement in cultured neurons from symptomatic mutant HSPB1 mice. 3. For compounds showing the best effects in Aim 2, further characterize these compounds by determining their experimental ADMET parameters in order to ensure appropriate stability, cell permeability, lack of off- target activity, safety, etc. 4. Lastly, tes the best compounds from Aim 3 in the CMT transgenic mouse models, characterizing these as we have done before on their capacity to enhance tubulin acetylation in peripheral nerves and to reverse the symptoms in the mutant HSPB1 mice.

描述(申请人提供)：Charcot-Marie-Tooth病(CMT)是最常见的遗传性神经疾病之一，在美国每2500人中就有1人受到影响。这种疾病是以1886年首次发现这种疾病的三位医生的名字命名的。肌萎缩侧索硬化症，也被称为遗传性运动和感觉神经病(HMSN)或腓肠肌萎缩，由一组影响周围神经的疾病组成。CMT同时影响运动神经和感觉神经。典型的特征包括足部和小腿肌肉无力，这可能导致脚部下垂和高台阶步态，经常绊倒或摔倒。小分子热休克蛋白27(HSPB1)的突变可引起轴索CMT或远端遗传性运动神经病(远端HMN)。我们的团队已经开发出并鉴定了仅在神经元中表达突变(S135F)HSPB1的转基因小鼠。这些小鼠表现出CMT2的所有特征。突变型HSPB1的表达降低了乙酰化微管蛋白水平，并导致严重的轴突运输缺陷。具有重要意义的是，我们现在已经证明，使用我们设计的HDAC6I[HDAC6抑制剂]Tubasatin A，药物抑制组蛋白脱乙酰酶6(HDAC6)诱导的微管蛋白脱乙酰化，纠正了HSPB1突变引起的轴突运输缺陷，并挽救了症状突变的HSPB1小鼠的CMT表型。我们的发现证明了微管蛋白脱乙酰化在突变的HSPB1诱导的神经病变中的致病作用，并为HDAC6抑制剂作为遗传性轴索病变的治疗策略提供了有价值的前景。为了扩大这些高影响力的发现，并将最好的HDAC6I疗法推向临床，治疗CMT和相关的神经疾病，我们打算更全面地探索Tubasatin A类似物家族的结构和活性关系[SAR]，Tubasatin A类似物是我们研究中使用的关键HDAC抑制剂。本研究的目的如下：1.利用药物化学原理和计算机模拟方法，设计和合成改良的Tubasatin A类似物，并进一步优化其体外HDAC异构体的选择性和效价。为了达到这个目的，在电子计算机中，将进行ADMET计算，以便只为初步的体外研究准备最类似药物的化合物。2.对于那些对HDAC6的选择性至少是其他异构体的300倍并且IC50为50 nM的化合物，优化这些新的化学实体以增强其在有症状的突变HSPB1小鼠的培养神经元中促进微管蛋白乙酰化和线粒体运动的能力。3.对于在Aim 2中表现出最好效果的化合物，通过测定它们的实验ADMET参数来进一步表征这些化合物，以确保适当的稳定性、细胞渗透性、缺乏靶外活性、安全性等。4.最后，在CMT转基因小鼠模型中TES来自Aim 3的最好的化合物，像我们以前所做的那样表征它们在增强周围神经微管蛋白乙酰化和扭转突变的HSPB1小鼠症状方面的能力。