Drug Discovery for Duchenne Muscular Dystrophy via Utrophin Promoter Activation Screening
Drug Discovery for Duchenne Muscular Dystrophy via Utrophin Promoter Activation Screening
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DOI：
10.1371/journal.pone.0026169
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发表时间：
2011-10-20
期刊：
影响因子：
3.7
通讯作者：
Khurana, Tejvir S.
中科院分区：
文献类型：
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作者：
Moorwood, Catherine;Lozynska, Olga;Khurana, Tejvir S.
背景：杜氏肌营养不良症 (DMD) 是一种毁灭性的肌肉消耗性疾病，由肌营养不良蛋白（一种肌肉细胞骨架蛋白）突变引起。 Utropin 是肌营养不良蛋白的同源物，当肌纤维中表达水平升高时，可以在功能上补偿肌营养不良蛋白的缺失，如肌营养不良蛋白缺陷小鼠的研究所示。因此，Utropin 上调是一种很有前景的 DMD 治疗方法。使用类似药物的小分子来实现肌营养蛋白上调在递送和生物利用度方面具有明显的优势。此外，通过筛选已批准用于临床的分子，可以消除开发新药所需的大部分时间和费用。方法/主要发现：我们开发并验证了一种基于细胞的、高通量的 utropin 启动子激活筛选方法，并用它来筛选市售药物和天然化合物的 Prestwick 化学库。初步筛选产生了 20 个命中分子，其中 14 个表现出 utropin 启动子的剂量依赖性激活，并被确认为命中分子。独立验证表明，其中一种化合物萘丁美酮能够上调 C2C12 肌肉细胞中的内源性肌营养不良蛋白 mRNA 和蛋白质。 结论/意义：我们开发了一种基于细胞的高通量筛选肌营养不良蛋白启动子测定法。使用该测定，我们鉴定并验证了一种utropin启动子激活药物nabumetone，其在人体中的药代动力学和安全性已经得到很好的描述，并且它代表了utropin上调作为DMD治疗的先导化合物。
Background: Duchenne muscular dystrophy (DMD) is a devastating muscle wasting disease caused by mutations in dystrophin, a muscle cytoskeletal protein. Utrophin is a homologue of dystrophin that can functionally compensate for its absence when expressed at increased levels in the myofibre, as shown by studies in dystrophin-deficient mice. Utrophin upregulation is therefore a promising therapeutic approach for DMD. The use of a small, drug-like molecule to achieve utrophin upregulation offers obvious advantages in terms of delivery and bioavailability. Furthermore, much of the time and expense involved in the development of a new drug can be eliminated by screening molecules that are already approved for clinical use.Methodology/Principal Findings: We developed and validated a cell-based, high-throughput screening assay for utrophin promoter activation, and used it to screen the Prestwick Chemical Library of marketed drugs and natural compounds. Initial screening produced 20 hit molecules, 14 of which exhibited dose-dependent activation of the utrophin promoter and were confirmed as hits. Independent validation demonstrated that one of these compounds, nabumetone, is able to upregulate endogenous utrophin mRNA and protein, in C2C12 muscle cells.Conclusions/Significance: We have developed a cell-based, high-throughput screening utrophin promoter assay. Using this assay, we identified and validated a utrophin promoter-activating drug, nabumetone, for which pharmacokinetics and safety in humans are already well described, and which represents a lead compound for utrophin upregulation as a therapy for DMD.