噻唑-噁唑串联杂环类RNA剪接抑制剂的发现及构效关系研究

doi:10.6023/cjoc202202033

摘要/Abstract

剪接是将内含子从前体mRNA (pre-mRNA)中移除, 并将侧翼外显子连接在一起形成成熟的mRNA的过程, pre-mRNA剪接过程是高等生物中基因表达和蛋白质组学多样性的重要组成部分, 但其发生选择性剪接或剪接因子发生突变, 都会导致疾病的发生. 利用小分子化合物对剪接过程的调控, 或为相关疾病的治疗提供另一种选择. 利用circmCherry表达体系进行化合物筛选, 发现天然产物Leucamide A的简单衍生物对于剪接过程具有一定的抑制作用, 以其为先导化合物, 进行了结构改造后, 新设计合成了约36个噻唑-噁唑双杂环串联结构的化合物, 其中2-[(S)-1- (2-{2-[(S)-1-(4-甲氧基苯甲酰胺)-2-甲基丙基]噻唑-4-基}-5-甲基噁唑-4-甲酰胺基)乙基]噁唑-4-甲酸甲酯(9l)对于剪接过程有较强的抑制作用.

关键词: 剪接抑制剂, circmCherry表达体系, Leucamide A衍生物, 构效关系

Precursor mRNAs (pre-mRNAs) undergo splicing to remove introns and ligation of their exons to yield mature mRNAs. Pre-mRNA splicing is an essential component of gene expression and proteomic diversity in higher organisms. However, the alternative splicing or mutation of splicing factors have been found in human diseases. By the use of the small molecules modulate the splicing process will provide potential treatment for related diseases. In this paper, circmcherry expression system was used to screen compounds that could affect splicing process. Several simple derivatives of natural product Leucamide A have been found to have inhibitory effects, and 36 compounds with thiazole-oxazole tadem structural core were designed and synthesized, of which methyl 2-((S)-1-(2-(2-((S)-1-(4-methoxybenzamido)-2-methylpropyl)thiazol-4- yl)-5-methyloxazole-4-carboxamido)ethyl)oxazole-4-carboxylate (9l) shows most potent inhibitory activity on splicing.

Key words: splicing inhibitors, circmCherry-expression system, Leucamide A derivatives, structure-activity relationships

引用此文

张蓉, 郜祥, 陈玲玲, 南发俊. 噻唑-噁唑串联杂环类RNA剪接抑制剂的发现及构效关系研究[J]. 有机化学, 2022, 42(9): 2925-2939.

Rong Zhang, Xiang Gao, Lingling Chen, Fajun Nan. Discovery and Structure-Activity Relationship Studies of Thiazole- Oxazole Tandem Heterocyclic RNA Splicing Inhibitors[J]. Chinese Journal of Organic Chemistry, 2022, 42(9): 2925-2939.

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图/表 12

图1. 线性剪接抑制剂

Figure 1. Liner splicing inhibitors

图2. 活性化合物的筛选

Figure 2. Screening of active compounds

图3. Leucamide A衍生物的设计思路

Figure 3. Design idea of Leucamide A derivatives

图式1. Leucamide A衍生物的合成

Scheme 1. Synthesis of Leucamide A derivatives

图4. W3/W11/W12 细胞毒性及量-效关系

Figure 4. Cytotoxicity and dose-effect relationship of W3/W11/W12

图5. R1基团的考察

Figure 5. Investigation of R1 group

图6. 去除异丙基侧链后的活性

Figure 6. Activity after isopropyl removal

图7. 酯基水解后的活性

Figure 7. Activity after ester hydrolysis

图8. 噁唑环的去除及替换后的活性

Figure 8. Activity after removal and replacement of oxazole

图9. 化合物9l的细胞毒性及其在给药浓度为20 μmol/L时的活性结果

Figure 9. Cytotoxicity of compound 9l and its activity results at dose concentration of 20 μmol/L

图10. 噻唑-噁唑双环类剪接抑制剂的构效关系

Figure 10. Structure-activity relationship of thiazole-oxazole tadem heterocyclic RNA splicing inhibitors

Primer	Sequence
qEGFP-F	GACCACATGAAGCAGCACGA
qEGFP-R	TGAAGTCGATGCCCTTCAG
qCircmCherry-F	TGCGCTTCAAGGTGCACATG
qCircmCherry-R	ACAGGATGTCCCAGGCGAAG
qEGFP-mCherry-F	GACCACATGAAGCAGCACGA
qEGFP-mCherry-R	ACAGGATGTCCCAGGCGAAG

表1. 筛选报告基因的引物

Table 1. Primers for screening reporter gene

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