非甾类FXR配体设计优化及调控机制研究与两种优势骨架的合成方法学研究

发布时间：2018-06-12 03:16

本文选题：基于药效团拼合原理的药物设计 + FXR配体　；参考：《华东理工大学》2015年博士论文

【摘要】：本论文由两个部分组成：第一部分是基于FXR配体的设计优化及调控机制研究。FXR是核受体家族中的一员,主要在肝脏、肠、肾脏以及肾上腺等器官中表达。目前,FXR已经明确能够在多种代谢途径中起到调节作用,包括胆固醇、胆汁酸、脂类以及葡萄糖代谢。此外,FXR与胆汁淤积、动脉粥样硬化、胆结石、糖尿病和癌症等多种疾病有着紧密的联系。然而目前的研究表明甾体类FXR配体在靶点选择性和基因选择性上均存在潜在的缺陷,设计非甾类的FXR配体对于设计开发以FXR为靶点的治疗药物具有重要意义。因此,我们探讨了非甾类FXR配体的设计优化以及调控机制研究。在前期自主开发的非甾类FXR拮抗剂(18)与非甾类FXR激动剂GW4064(24)的基础上,我们根据药效团拼合原理将GW4064(24)的疏水药效团与化合物18进行了拼合。结合化合物的可合成性和结合活性,选取了化合物36作为FXR配体的先导结构进行优化和探索,通过合理药物设计和化学合成手段,进行三轮结构改造,得到45个化合物(36,A1-19,B1-25)。通过测试其分子水平及细胞水平活性,最终发现了三个FXR激动剂(A10,B2和B3)和一个FXR拮抗剂(B25)。之后我们以FXR拮抗剂B25为配体,利用分子动力学模拟探究配体对FXR的拮抗机制,拮抗剂B25的构象变化引起了FXR激动形式的H12构象改变,阻碍共激活因子的招募。我们的分子模拟结果提出了FXR拮抗机制重要的分子基础,FXR结构中H12螺旋上的氨基酸E467的构象是FXR拮抗调控的关键,这与之前文献中报道的机制一致。第二部分是两种优势骨架的合成方法学研究。我们发展了以二苯基酮亚胺为关键底物的两种3-取代-2-吲哚酮优势骨架合成方法,应用这些方法可以快速构建优势骨架分子,为药物筛选研究提供快捷的合成方法以及较好的分子库。其一,以各种取代靛红(56)、甘氨酸叔丁酯盐酸盐(57)及二苯基酮亚胺(58)为原料,在温和条件下经一步三组分反应和一步辅助基团脱去,高效制得含3-羟基-2-吲哚酮类优势骨架的非天然氨基酸,产率58-90%。其二,以二苯基酮亚胺(58)为氨基供体,在含碘添加剂与TBHP的氧化体系下,高效完成醛类、二芳基甲烷类以及3-取代-2-吲哚酮类底物的氨基化反应,制得文献中极少报道的伯胺产物,产率38-89%。综上所述,基于药效团拼合原理,针对FXR这一与多种疾病密切相关的重要靶标,我们设计并合成了45个化合物(36,A1-19,B1-25),最终发现了该靶标的三个激动剂和一个拮抗剂,并首次利用自主研发的非甾类FXR拮抗剂进行了分子动力学模拟以解释FXR拮抗机制。另外,发展了以二苯基酮亚胺为关键底物的两种优势骨架合成方法,快速构建了两类3-取代-2-吲哚酮类优势分子骨架。这些研究为我们开发新药奠定了基础。
[Abstract]:This thesis consists of two parts: the first part is based on the design optimization and regulation mechanism of FXR ligand. FXR is a member of nuclear receptor family, mainly expressed in liver, intestine, kidney and adrenal gland. FXR has been identified to play a regulatory role in a variety of metabolic pathways, including cholesterol, bile acids, lipids, and glucose metabolism. In addition, FXR is closely associated with many diseases, such as cholestasis, atherosclerosis, gallstone, diabetes and cancer. However, current studies have shown that steroidal FXR ligands have potential defects in targeting selectivity and gene selectivity, and the design of non-steroidal FXR ligands is of great significance in the design and development of FXR targeted therapeutic drugs. Therefore, we discussed the design optimization and regulatory mechanism of nonsteroidal FXR ligands. On the basis of the previously developed non-steroidal FXR antagonist (C18) and the nonsteroidal FXR agonist (GW40644N24), the hydrophobic pharmacophore of GW406424) was combined with compound 18 according to the principle of pharmacophore assembly. Combined with the synthesis and binding activity of the compounds, compound 36 was selected as the leading structure of FXR ligands for optimization and exploration. Through rational drug design and chemical synthesis, 45 compounds were obtained by three-wheeled structural modification. Three FXR agonists, A10B _ 2 and B _ 3) and a FXR antagonist, B25, were found by measuring their molecular and cellular activity. Then we used FXR antagonist B25 as ligand to explore the antagonistic mechanism of ligand against FXR by molecular dynamics simulation. The conformation change of antagonist B25 caused the conformation change of FXR excited form H12, which hindered the recruitment of coactivator. Our molecular simulation results suggest that the conformation of the amino acid E467 on the H12 helix in the FXR structure is the key to the regulation of FXR antagonism, which is consistent with the previously reported mechanism. The second part is the synthesis methodology of two dominant skeletons. We have developed two methods for the synthesis of 3-substituted -2-indole ketone dominant skeleton using benzophenone imine as the key substrate, which can be used to construct the dominant skeleton molecule quickly. To provide a rapid synthetic method and a better molecular library for drug screening. First, with various substituted indirubin 56, glycine tert-butyl hydrochloride 57) and benzophenone imine 58) as raw materials, under mild conditions, they were removed by one step, three component reaction and one step auxiliary group. An unnatural amino acid containing 3-hydroxyl-2-indoleones was prepared in 58-90 yield. Secondly, the amino reactions of aldehydes, diaryl methane and 3-substituted -2-indole ketone substrates were efficiently completed in the oxidation system containing iodine and TBHP, using benzophenone iminimide 58) as the amino donor, and in the oxidation system containing iodine and TBHP, the amino reactions of aldehydes, diaryl methane and 3-substituted -2-indole ketones were efficiently completed. The primary amines, which are rarely reported in the literature, have been prepared in 38-89. To sum up, based on the principle of pharmacophore assembly, we designed and synthesized 45 compounds, C36A1-19OB1-25N, for FXR, an important target closely related to many diseases, and finally found three agonists and an antagonist for this target. The molecular dynamics simulation of non-steroidal FXR antagonist was carried out for the first time to explain the mechanism of FXR antagonism. In addition, two kinds of dominant skeleton synthesis methods with benzophenone imine as the key substrate were developed, and two kinds of 3-substituted -2-indole ketones dominant molecular skeletons were constructed rapidly. These studies have laid the foundation for the development of new drugs.
【学位授予单位】：华东理工大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R914

【相似文献】