Design, Optimization, and Study of Small Molecules That Target Tau Pre-mRNA and Affect Splicing
Design, Optimization, and Study of Small Molecules That Target Tau Pre-mRNA and Affect Splicing
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DOI：
10.1021/jacs.0c00768
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发表时间：
2020-05-13
影响因子：
15
通讯作者：
Disney, Matthew D.
中科院分区：
文献类型：
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作者：
Chen, Jonathan L.;Zhang, Peiyuan;Disney, Matthew D.
大约95%的人类基因是选择性剪接的，异常的剪接事件可引起疾病。一种被选择性剪接并与神经退行性疾病相关的前mrna是tau（微管相关蛋白tau），它可以导致与17号染色体相关的额颞叶痴呆和帕金森病（FTDP-17），并可能导致阿尔茨海默病。在这里，我们描述了结构特异性先导小分子的设计，直接从序列中靶向tau前mrna。随后进行了热膨胀和类似物合成，以进一步改进这些初始铅分子。这些新化合物通过一系列试验评估其功能活性，包括结合试验和通过U1小核核糖核蛋白（snRNP）剪接因子模拟tau前mrna的分子识别试验。从这些研究中产生的化合物相对于最初的目标RNA具有增强的效力和选择性，同时也具有显着改善的药物样特性。这些化合物被证明可以通过化学交联和下拉目标谱分离直接靶向细胞中的tau前mrna，并在包括初级神经元在内的多种细胞系统中挽救tau前mrna的疾病相关剪接。更广泛地说，这项研究表明，可以从序列设计铅，结构特异性化合物，然后进一步优化其物理化学性质，同时增强其活性。
Approximately 95% of human genes are alternatively spliced, and aberrant splicing events can cause disease. One pre-mRNA that is alternatively spliced and linked to neurodegenerative diseases is tau (microtubule-associated protein tau), which can cause frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17) and can contribute to Alzheimer's disease. Here, we describe the design of structure-specific lead small molecules that directly target tau pre-mRNA from sequence. This was followed by hit expansion and analogue synthesis to further improve upon these initial lead molecules. The emergent compounds were assessed for functional activity in a battery of assays, including binding assays and an assay that mimics molecular recognition of tau pre-mRNA by a U1 small nuclear ribonucleoprotein (snRNP) splicing factor. Compounds that emerged from these studies had enhanced potency and selectivity for the target RNA relative to the initial hits, while also having significantly improved drug-like properties. The compounds are shown to directly target tau pre-mRNA in cells, via chemical cross-linking and isolation by pull-down target profiling, and to rescue disease-relevant splicing of tau pre-mRNA in a variety of cellular systems, including primary neurons. More broadly, this study shows that lead, structure-specific compounds can be designed from sequence and then further optimized for their physicochemical properties while at the same time enhancing their activity.