GPR120小分子荧光探针及其激动剂的研究

发布时间：2018-04-06 21:06

本文选题：GPR120　切入点：GPCR　出处：《山东大学》2017年硕士论文

【摘要】：GPR120 受体(G protein-coupled receptor 120)是 G蛋白偶联受体(GPCRs)大家族的一员,广泛地分布于肺、结肠和脂肪等组织中,具有调节GLP-1分泌、胰岛素敏感性、抗炎和抗肥胖等多种生理学功能,且与许多生理疾病存在一定关系,例如2型糖尿病、饮食、肥胖等。因此,采用小分子作为工具进行GPR120受体的系统生理学及病理学研究对于糖尿病和肥胖的发生机制和治疗具有至关重要的意义。本论文主要包括两部分内容:设计评价了七个用于识别GPR120受体的小分子荧光探针;设计并评价了一系列GPR120受体的小分子激动剂。(1)GPR120受体的小分子荧光探针的研究这些年来,具有荧光性质的GPCR配体已经广泛应用于定位受体的分布,并且能够实时监测由配体-受体相互作用触发的过程(例如内化、运输、螯合和回收)。然而,迄今为止还没有关于检测GPR120受体在细胞表面分布的小分子荧光探针的报道。鉴于小分子荧光探针具有高灵敏度,选择性,可视化和快速响应等优点,迫切需要开发一种方便的荧光配体来追踪GPR120,以深入理解GPR120生理学和病理学功能作用。小分子荧光探针一般有3部分构成:荧光团、连接基团以及识别基团。本论文选取GPR120的激动剂GW9508和TUG-891与靶点结合位点的母核结构作为GPR120药效团模型进行结构改造,以萘二酰亚胺、香豆素作为等具有出色光学特性的化合物作为荧光团,并通过烷基链进行化学链接,以期得到高灵敏度、高选择性的GPR120受体小分子荧光探针,从而对GPR120受体的生物学及药理学研究提供一定帮助。最终,我们设计并评价了七个识别GPR120受体的小分子荧光探针。随后对合成的GPR120荧光探针进行了光学特性的测定、BRET活性实验、钙离子实验测定、细胞成像研究以及细胞毒性的研究。结果显示,这些荧光探针对GPR120受体表现出优良的生物活性、良好的选择性、可接受的细胞毒性以及成功应用于细胞荧光成像研究。因此,荧光探针可以作为标记GPR120的染色工具,提供详细的GPR120的生理学和病理学功能信息。此外,我们期望这些探针可以用作GPR120配体活性筛选中的荧光竞争性底物,从而构建快速筛选激动剂化合物平台。(2)GPR120受体的小分子激动剂的研究已有报道显示,GPR120激动作用可通过调节脂肪形成、胃肠肽分泌、味道偏好和葡萄糖代谢平衡来改善机体炎症、糖尿病等相关代谢健康。因此,开发GPR120的激动剂将作为一种新的治疗选择,可用于治疗受损的代谢疾病,如肥胖症、2型糖尿病和心血管疾病等。本课题中,合成并筛选了一系列优秀的GPR120芳香酸类小分子激动剂,期望能对GPR120受体相关代谢疾病的治疗提供帮助,如炎症、肥胖症和2型糖尿病等。本课题通过筛选一系列GPR120受体合成的激动剂,化合物A11被鉴定为有效的GPR120激动剂,与阳性药TUG-891表现出相似的分子机制,可导致细胞内钙离子响应,β-arrestin2募集以及受体的快速内在化。此外,我们期望化合物11能够作为一个重要的药理学工具,去探索GPR120受体的相关生物学功能,以达到治疗肥胖症、2型糖尿病及其他其他代谢疾病的目的。综上所述,本课题得到的GPR120小分子荧光探针及其激动剂,将为GPR120受体的相关研究提供新的研究工具,也为今后此类荧光探针或激动剂的进一步结构改造提供理论指导。
[Abstract]:GPR120 receptor (G protein-coupled receptor 120) is a G protein coupled receptor (GPCRs) is a member of a large family, widely distributed in the lung, colon and adipose tissue, regulate the secretion of GLP-1, insulin sensitivity, anti-inflammatory and anti obesity function of various physiological, and there is a certain relationship with many physiological diseases, such as 2 diabetes, diet, obesity and so on. Therefore, the study has an important significance for the pathogenesis and treatment of obesity and diabetes in the physiology and pathology of the system with small molecules as a tool for GPR120 receptor. This paper consists of two parts: the design and evaluation of the seven for small molecule fluorescent probe recognition GPR120 receptor; design and the evaluation of a series of small molecule agonists of the GPR120 receptor. (1) research on small molecule fluorescent probe of the GPR120 receptor in these years, GPCR ligand with fluorescence has been widely For the localization of the receptor, and can process real-time monitoring triggered by ligand receptor interactions (e.g., internalization, transport, chelation and recovery). However, so far there is no report about the detection of GPR120 receptor in small molecule fluorescent probe distribution of cell surface. In view of the small molecule fluorescent probe with high sensitivity and selectivity. The visualization and the advantages of rapid response, there is an urgent need to develop a convenient fluorescent ligand to track GPR120, to GPR120 understanding the physiological and pathological functions. Small molecule fluorescent probe generally consists of 3 parts: fluorophores, linkers and recognition group. As the GPR120 pharmacophore model nulei structure this thesis chooses GPR120 agonist GW9508 and TUG-891 and target binding sites for structural transformation, with two naphthalene imide compounds, coumarin as having excellent optical properties as fluorescence Group, and the chemical linked by alkyl chain, in order to obtain high sensitivity, GPR120 receptor small molecule fluorescent probe with high selectivity, and biological and pharmacological research of GPR120 receptor provides a help. Finally, we design and evaluate small molecule fluorescent probe seven identified GPR120 receptor. Followed by determination of optical properties the GPR120 fluorescence probe, BRET activity test, determination of calcium ion on experiment, cell imaging and cell toxicity. The results showed that the fluorescent probe of GPR120 receptor showed excellent biological activity, good selectivity, cytotoxicity of acceptable and successful applications on cell fluorescence imaging. Therefore, fluorescence probe can be used as a marker for GPR120 staining tool, GPR120 provides detailed physiological and pathological information. In addition, we expect these probes can be used as GPR120 Ligand activity screening of competitive fluorescent substrate, so as to construct the platform for rapid screening of agonist compounds. (2) small molecule agonists of the GPR120 receptor research has been reported, the GPR120 agonist can be formed by regulating fat, gastrointestinal peptide secretion, taste preference and glucose metabolism to improve inflammation, diabetes and other related metabolism health. Therefore, the development of GPR120 agonists will serve as a new therapeutic option for treatment of impaired metabolic diseases, such as obesity, type 2 diabetes and cardiovascular disease. In this paper, the synthesis and screening of a series of excellent GPR120 aromatic acids, small molecule agonists, is expected to the treatment of GPR120 receptor related metabolic diseases help, such as inflammation, obesity and type 2 diabetes. This topic through the screening of a series of synthetic GPR120 agonists, compound A11 was identified as a valid GPR120 Agonist, and positive drug TUG-891 showed similar molecular mechanisms that can lead to intracellular calcium response, rapid internalization of beta -arrestin2 recruitment and receptor. In addition, we expect 11 compounds can be used as an important pharmacological tool to explore the related biological function of the GPR120 receptor, in order to achieve the treatment of obesity, type 2 diabetes and other metabolic diseases. In summary, GPR120 small molecule fluorescent probe and supported by the agonist, will provide a new research tool for the study of the GPR120 receptor, but also provide theoretical guidance for the further structural transformation in such fluorescent probe or agonist.

【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R914

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