Pediatric toxicity and efficacy in long-term systemic treatment with anti-sense

反义药物长期全身治疗的儿科毒性和疗效

• 批准号: 8246746
• 负责人: JOHANNES NICOLAAS VAN DEN ANKER
• 金额: $ 87.62万
• 依托单位: CHILDREN'S RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-26 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8246746
• 关键词: Pediatric; toxicity; efficacy; long; term

DESCRIPTION (provided by applicant): Duchenne muscular dystrophy is the most common monogenic pediatric inborn error, affecting one in 3,500 live born males world-wide. The most promising molecular therapeutic approach for DIVID is systemic delivery of anti-sense oligonucleotides (AOs), where the drugs alter mRNA splicing, and converting out-of-frame loss-off function transcripts to in-frame transcripts capable of producing semi-functional (Becker-like) dystrophin. In the large animai (dog) model of Duchenne dystrophy, we have shown that intravenous (iV) delivery of high dose morpholino AOs is able to rescue dystrophin protein production to about 20% of wild-type levels, and cause stabilization or improvement of multiple functional, histological, and imaging outcome measures (Yokota et al. 2009). Initial human clinical trials using morpholino AO directed at exon 51 have shown that both direct intramuscular injection and IV administration can result in de novo dystrophin production. However, it is anticipated that repeated doses of 20 mg/kg - 100 mg/kg are likely required for sustained efficacy. GLP toxicity studies in mice and non-human primates, sponsored by the applicants, have shown that high dose weekly IV injections can lead to accumulation of drug in kidney proximal tubule cells. This accumulation resolved after termination of dosing, and there were no elevation of standard markers of kidney damage. However, dosing of DIVID patients will need to be life-long, and there has been no optimization of dosing schedules or drug concentrations that balance efficacy in muscle vs. kidney accumulation. Here, we bring together an interdisciplinary team to define the therapeutic window of morpholino dosing. Project 1 collaborates with an ongoing dose-ranging clinical trial to monitor kidney toxicity through urine biomarkers and shed renal cells. Project 2 defines the effects of AO concentrations, and dosing regimen on drug accumulation in kidney tubule cells, and also carries out a biomarker discovery program to define sensitive and reliable urine biomarkers for morpholino-associated cell damage. Project 3 defines the optimal dosing regimen able to provide sustained clinical efficacy in the mouse model of muscular dystrophy, using an established murine drug-testing core. A kidney toxicology assessment core supports these projects. PUBLIC HEALTH RELEVANCE: This project will provide clinical samples to monitor kidney accumulation of morpholino AO with chronic high dose IV treatment in boys with DMD. This project will integrate the pre-clinical and clinical data from the entire U54 program to derive inform optimized dosing and therapeutic index. The treatment of DMD using high dose IV morpholino is the most promising strategy for therapeutics of DMD. The possible consequences of long-term chronic treatment are not known. The data from this Project 2 will help define intervention targets and test tools for effective treatment of DMD using antisense morpholino. We will determine an optimized therapeutic window using the mouse efficacy and rat toxicity data above, including renal function tests, renal histology and ultrastructure, urine epithelial cel assays, and urine proteomic biomarker assays through Project 2 and Core B. Use of PMOs to correct genetic defects is rapidly advancing to several other human diseases such as FSHD, Cystic fibrosis, myotonic dystrophy, LGMD etc. Therefore, the data generated from this proposal have much broader implications than Duchenne muscular dystrophy.

描述（由申请人提供）：杜兴氏肌营养不良症是最常见的单基因儿科先天性缺陷，影响全球每 3,500 名活产男性中的一名。 DIVID 最有前途的分子治疗方法是系统性递送反义寡核苷酸 (AO)，其中药物改变 mRNA 剪接，并将框外丧失功能的转录物转化为能够产生半功能性的框内转录物。 （类似贝克尔） 肌营养不良蛋白。在杜氏营养不良的大型动物（狗）模型中，我们已经证明静脉（iV）输送高剂量 剂量吗啉代 AO 能够将抗肌营养不良蛋白产量恢复到野生型水平的约 20%，并导致多种功能、组织学和成像结果指标的稳定或改善（Yokota 等，2009）。使用针对外显子 51 的吗啉代 AO 进行的初步人体临床试验表明，直接肌内注射和静脉注射均可导致肌营养不良蛋白从头产生。然而，预计可能需要 20 mg/kg - 100 mg/kg 的重复剂量才能获得持续功效。由申请人赞助的小鼠和非人类灵长类动物的 GLP 毒性研究表明，每周高剂量静脉注射可导致药物在肾近曲小管细胞中积聚。这种积累在给药终止后消失，并且肾损伤的标准标志物没有升高。然而，DIVID 患者的给药需要终生进行，并且尚未优化给药方案或药物浓度来平衡肌肉与肾脏蓄积的功效。在这里，我们聚集了一个跨学科团队来定义吗啉剂量的治疗窗口。项目 1 与一项正在进行的剂量范围临床试验合作，通过尿液生物标志物和脱落的肾细胞监测肾毒性。项目 2 定义了 AO 浓度和给药方案对肾小管细胞药物蓄积的影响，并实施了生物标志物发现计划，以确定吗啉代相关细胞损伤的敏感且可靠的尿液生物标志物。项目 3 使用已建立的小鼠药物测试核心，定义了能够在肌营养不良小鼠模型中提供持续临床疗效的最佳给药方案。肾脏毒理学评估核心支持这些项目。 公共健康相关性：该项目将提供临床样本，以监测患有 DMD 的男孩接受长期高剂量静脉注射治疗后肾脏中吗啉代 AO 的积累情况。该项目将整合整个 U54 项目的临床前和临床数据，以获得优化剂量和治疗指数的信息。使用高剂量静脉注射吗啉治疗 DMD 是最有前途的 DMD 治疗策略。长期慢性治疗可能产生的后果尚不清楚。该项目 2 的数据将有助于确定使用反义吗啉有效治疗 DMD 的干预目标和测试工具。我们将利用上述小鼠疗效和大鼠毒性数据确定优化的治疗窗口，包括肾功能测试、肾脏组织学和超微结构、尿液上皮细胞测定和尿液蛋白质组生物标志物测定 通过项目 2 和核心 B。使用 PMO 来纠正遗传缺陷正在迅速推进到其他几种人类疾病，如 FSHD、囊性纤维化、强直性肌营养不良、LGMD 等。因此，从该提案生成的数据比 Duchenne 肌肉疾病具有更广泛的影响营养不良。