哌啶胺类L3MBTL1拮抗剂的设计与合成

发布时间：2018-09-08 09:23

【摘要】：表观遗传学是由染色体改变所引起的稳定的可遗传的表现型，而非DNA序列的改变。进行表观遗传学研究的依据在于，尽管生物体内的所有细胞都继承了相同的遗传物质，但是DNA和染色质包装形式的可遗传性的差异导致了不同细胞具有不同的物理特性和生物学功能。近年来对表观遗传学的研究发现，，其对攻克癌症、感染、代谢系统疾病等长期困扰人类的难题具有重大意义。组蛋白的共价修饰和DNA的甲基化是影响表观基因改变的两个主要原因。作为组蛋白修饰的一种，组蛋白中赖氨酸甲基化状态的每一点细微变化都会对蛋白质的相互作用以及基因的调控和表达产生巨大影响，所以对组蛋白中赖氨酸残基甲基化状态的识别是染色质调控的重中之重。 MBT（恶性脑瘤The malignant brain tumor）域对赖氨酸甲基化有着特殊的选择性。其与甲基化的赖氨酸的独特的结合方式（嵌入式）为人工设计并合成一些选择性的配体提供了一定的可能性，这也是选择MBT域作为甲基赖氨酸拮抗剂研究的突破口的原因。MBT域中L3MBTL1对于基因组的稳定起到至关重要的作用，其能抑制骨髓功能失调引发的肿瘤，它的消耗能够促进人类造血细胞的红细胞分化。但是目前得到的L3MBTL1拮抗剂的活性还不够理想，需要进一步研究。本文依据L3MBTL1口袋的特点及化合物UNC669与L3MBTL1口袋的作用关系，在关键作用片段4-吡咯烷基-哌啶的基础上，考察了其类似物为作用片段对生物活性的影响。本课题共设计了三类化合物，分别是哌啶甲酮类化合物、嘧啶并吡咯类化合物及含有新型母核嘧啶并六氢嘧啶并四氢吡咯结构的化合物。其中，哌啶甲酮类化合物通过4条路线得到，共计6个化合物；嘧啶并吡咯类化合物通过1条路线得到，共计11个化合物；嘧啶并六氢嘧啶并四氢吡咯类化合物通过2条路线得到，共计8个化合物。本课题共采用了7条不同的合成路线，最终共得到25个目标化合物。在北卡罗来纳大学（UNC）Frye课题组的帮助下，采用同质邻近发光放大（AlphaScreen）和等温滴定微量热法（ITC）相结合的方法对化合物进行生物活性测试。结果显示，部分化合物对L3MBTL1具有一定的选择活性。目前，我们所合成的化合物中，对L3MBTL1活性最好的是嘧啶并吡咯类化合物2-29c（IC50=2.6±0.79μM），但其活性仅略优于UNC926（IC50=3.9±1.1μM），仍需继续研究。同时，对于具有新型母核嘧啶并六氢嘧啶并四氢吡咯类化合物，虽然只有化合物2-38a具有较弱的生物活性（IC50=38±3.2μM），但在此结果的基础上，对化合物进一步设计，可能得到活性更高的L3MBTL1拮抗剂。通过本课题对L3MBTL1拮抗剂的研究，我们对L3MBTL1口袋特点及其拮抗剂的设计有了进一步的认识，为今后寻找更高活性的L3MBTL1拮抗剂提供了参考。
[Abstract]:Epigenetics is a stable heritable phenotype caused by chromosome change, not a change in DNA sequence. Epigenetics is based on the fact that although all cells in the organism inherit the same genetic material, However, the genetic differences between DNA and chromatin packaging led to different physical properties and biological functions of different cells. In recent years, epigenetics has been found to be of great significance in solving the problems of cancer, infection, metabolic system diseases and so on. Covalent modification of histone and methylation of DNA are two main factors affecting epigenetic changes. As a kind of histone modification, every slight change of lysine methylation in histone has a great effect on protein interaction and gene regulation and expression. Therefore, the recognition of lysine residue methylation in histone is the most important part of chromatin regulation in. MBT (malignant brain tumor. The malignant brain tumor) domain has special selectivity for lysine methylation. Its unique binding to methylated lysine (embedded) provides a possibility for the artificial design and synthesis of some selective ligands. This is why the MBT domain is chosen as a breakthrough in the study of methyllysine antagonists. L3MBTL1 in the MBT domain plays a crucial role in the stability of the genome, which inhibits tumors caused by bone marrow dysfunction. Its consumption can promote erythrocyte differentiation of human hematopoietic cells. However, the activity of L3MBTL1 antagonist is not ideal, so it needs further study. Based on the characteristics of L3MBTL1 pocket and the interaction between compound UNC669 and L3MBTL1 pocket, the effects of its analogues on biological activity were investigated on the basis of 4-pyrrolidinyl-piperidine. In this paper, three kinds of compounds were designed, namely piperidone, pyrimidine pyrrole and pyrimidine hexahydropyrimidine tetrahydropyrrole. Among them, piperidone compounds were obtained by 4 routes, pyrimidine pyrrole compounds by one route, and pyrimidine pyrrole compounds by one route, with a total of 11 compounds. Pyrimidine hexahydropyrimidine tetrahydropyrrole compounds were obtained by two routes, a total of 8 compounds. In this study, 7 different synthetic routes were used and 25 target compounds were obtained. With the help of the (UNC) Frye team of the University of North Carolina, the bioactivity of the compounds was tested by means of isothermal titration microcalorimetry (ITC) and isomeric proximity luminescence amplification (AlphaScreen). The results showed that some of the compounds had selective activity to L3MBTL1. At present, the pyrimidine pyrrole compound 2-29c (IC50=2.6 卤0.79 渭 M),) has the best L3MBTL1 activity, but its activity is only slightly better than that of UNC926 (IC50=3.9 卤1.1 渭 M),). At the same time, although only compounds 2-38a have weak biological activity (IC50=38 卤3.2 渭 M), the compounds are further designed on the basis of the results, which have a novel parent pyrimidine hexahydropyrimidine tetrahydropyrrole compounds, while only compounds 2-38a have weak biological activity (IC50=38 卤3.2 渭 M),). A more active L3MBTL1 antagonist may be obtained. Through the study of L3MBTL1 antagonists, we have a further understanding of the characteristics of L3MBTL1 pocket and the design of its antagonists, which provides a reference for finding more active L3MBTL1 antagonists in the future.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：O621.13;R914.5

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