Optimization of HDAC6 Inhibitors in the Treatment of CMT

HDAC6 抑制剂治疗 CMT 的优化

• 批准号: 9035438
• 负责人: Ludo Van Den Bosch
• 金额: $ 38.41万
• 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 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9035438
• 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; HSPB1 gene; Hammer Toes; Hand; Health; Heat shock proteins; Hereditary Motor and Sensory Neuropathies; Hereditary Motor and Sensory Neuropathy Type I; Hereditary Motor and Sensory-Neuropathy Type II; Histone Deacetylase; Histone Deacetylase Inhibitor; In Vitro; Individual; Inherited; 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（由申请人提供）：腓骨肌萎缩症（CMT）是最常见的遗传性神经系统疾病之一，在美国每2,500人中就有1人患病。这种疾病是以1886年首次发现它的三位医生命名的。CMT，也称为遗传性运动和感觉神经病（HMSN）或腓骨肌萎缩症，是由一组影响周围神经的疾病组成。CMT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，这可能导致足下垂和频繁绊倒或福尔斯的高跨步步态。小热休克蛋白27（HSPB 1）的突变导致轴突CMT或远端遗传性运动神经病（远端HMN）。我们的团队已经开发并表征了仅在神经元中表达突变型（S135 F）HSPB 1的转基因小鼠。这些小鼠显示CMT 2的所有特征。突变型HSPB 1的表达降低乙酰化微管蛋白水平，并诱导严重的轴突运输缺陷。具有重要意义的是，我们现在已经表明，使用我们设计的HDAC 6 I [HDAC 6抑制剂] Tubastatin A对组蛋白脱乙酰酶6（HDAC 6）诱导的微管蛋白脱乙酰化的药理学抑制纠正了由HSPB 1突变诱导的轴突运输缺陷，并挽救了有症状的突变HSPB 1小鼠的CMT表型。我们的研究结果证明了微管蛋白脱乙酰化在突变型HSPB 1诱导的神经病变中的致病作用，并为HDAC 6抑制剂作为遗传性轴突病的治疗策略提供了有价值的前景。为了扩展这些高影响力的发现，并将最好的HDAC 6 I疗法推向临床，用于治疗CMT和相关神经病变，我们打算更充分地探索Tubastatin A类似物家族的结构活性关系[SAR]，Tubastatin A类似物是我们研究中使用的关键HDAC抑制剂。本建议的目的如下：1.使用药物化学原理和计算机建模方法，设计和合成Tubastatin A的改进类似物，并进一步优化它们的体外HDAC亚型选择性和效力。为此，将进行计算机模拟ADMET计算，以便仅制备最具药物样的化合物用于初步体外研究。2.对于那些对HDAC 6显示出超过其他同种型至少300倍的选择性并且具有< 50 nM的IC 50的化合物，优化这些新的化学实体以增强来自症状突变型HSPB 1小鼠的培养神经元中的微管蛋白乙酰化和线粒体运动的能力。3.对于在目标2中显示出最佳效果的化合物，通过确定其实验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、缺乏脱靶活性、安全性等。最后，在CMT转基因小鼠模型中测试来自Aim 3的最佳化合物，如我们之前所做的那样表征这些化合物增强外周神经中微管蛋白乙酰化和逆转突变HSPB 1小鼠中症状的能力。