本文選題：GPR120　切入點(diǎn)：GPCR　出處：《山東大學(xué)》2017年碩士論文

【摘要】：GPR120 受體(G protein-coupled receptor 120)是 G蛋白偶聯(lián)受體(GPCRs)大家族的一員,廣泛地分布于肺、結(jié)腸和脂肪等組織中,具有調(diào)節(jié)GLP-1分泌、胰島素敏感性、抗炎和抗肥胖等多種生理學(xué)功能,且與許多生理疾病存在一定關(guān)系,例如2型糖尿病、飲食、肥胖等。因此,采用小分子作為工具進(jìn)行GPR120受體的系統(tǒng)生理學(xué)及病理學(xué)研究對于糖尿病和肥胖的發(fā)生機(jī)制和治療具有至關(guān)重要的意義。本論文主要包括兩部分內(nèi)容:設(shè)計評價了七個用于識別GPR120受體的小分子熒光探針;設(shè)計并評價了一系列GPR120受體的小分子激動劑。(1)GPR120受體的小分子熒光探針的研究這些年來,具有熒光性質(zhì)的GPCR配體已經(jīng)廣泛應(yīng)用于定位受體的分布,并且能夠?qū)崟r監(jiān)測由配體-受體相互作用觸發(fā)的過程(例如內(nèi)化、運(yùn)輸、螯合和回收)。然而,迄今為止還沒有關(guān)于檢測GPR120受體在細(xì)胞表面分布的小分子熒光探針的報道。鑒于小分子熒光探針具有高靈敏度,選擇性,可視化和快速響應(yīng)等優(yōu)點(diǎn),迫切需要開發(fā)一種方便的熒光配體來追蹤GPR120,以深入理解GPR120生理學(xué)和病理學(xué)功能作用。小分子熒光探針一般有3部分構(gòu)成:熒光團(tuán)、連接基團(tuán)以及識別基團(tuán)。本論文選取GPR120的激動劑GW9508和TUG-891與靶點(diǎn)結(jié)合位點(diǎn)的母核結(jié)構(gòu)作為GPR120藥效團(tuán)模型進(jìn)行結(jié)構(gòu)改造,以萘二酰亞胺、香豆素作為等具有出色光學(xué)特性的化合物作為熒光團(tuán),并通過烷基鏈進(jìn)行化學(xué)鏈接,以期得到高靈敏度、高選擇性的GPR120受體小分子熒光探針,從而對GPR120受體的生物學(xué)及藥理學(xué)研究提供一定幫助。最終,我們設(shè)計并評價了七個識別GPR120受體的小分子熒光探針。隨后對合成的GPR120熒光探針進(jìn)行了光學(xué)特性的測定、BRET活性實(shí)驗、鈣離子實(shí)驗測定、細(xì)胞成像研究以及細(xì)胞毒性的研究。結(jié)果顯示,這些熒光探針對GPR120受體表現(xiàn)出優(yōu)良的生物活性、良好的選擇性、可接受的細(xì)胞毒性以及成功應(yīng)用于細(xì)胞熒光成像研究。因此,熒光探針可以作為標(biāo)記GPR120的染色工具,提供詳細(xì)的GPR120的生理學(xué)和病理學(xué)功能信息。此外,我們期望這些探針可以用作GPR120配體活性篩選中的熒光競爭性底物,從而構(gòu)建快速篩選激動劑化合物平臺。(2)GPR120受體的小分子激動劑的研究已有報道顯示,GPR120激動作用可通過調(diào)節(jié)脂肪形成、胃腸肽分泌、味道偏好和葡萄糖代謝平衡來改善機(jī)體炎癥、糖尿病等相關(guān)代謝健康。因此,開發(fā)GPR120的激動劑將作為一種新的治療選擇,可用于治療受損的代謝疾病,如肥胖癥、2型糖尿病和心血管疾病等。本課題中,合成并篩選了一系列優(yōu)秀的GPR120芳香酸類小分子激動劑,期望能對GPR120受體相關(guān)代謝疾病的治療提供幫助,如炎癥、肥胖癥和2型糖尿病等。本課題通過篩選一系列GPR120受體合成的激動劑,化合物A11被鑒定為有效的GPR120激動劑,與陽性藥TUG-891表現(xiàn)出相似的分子機(jī)制,可導(dǎo)致細(xì)胞內(nèi)鈣離子響應(yīng),β-arrestin2募集以及受體的快速內(nèi)在化。此外,我們期望化合物11能夠作為一個重要的藥理學(xué)工具,去探索GPR120受體的相關(guān)生物學(xué)功能,以達(dá)到治療肥胖癥、2型糖尿病及其他其他代謝疾病的目的。綜上所述,本課題得到的GPR120小分子熒光探針及其激動劑,將為GPR120受體的相關(guān)研究提供新的研究工具,也為今后此類熒光探針或激動劑的進(jìn)一步結(jié)構(gòu)改造提供理論指導(dǎo)。
[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.

【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R914

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本文編號：1718895