本文選題：PI3K/mTOR　切入點(diǎn)：免疫抑制劑　出處：《重慶理工大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：免疫抑制劑(immunosuppressant)是一類通過抑制細(xì)胞及體液免疫反應(yīng)而使組織損傷得以減輕的物質(zhì)。它可抑制機(jī)體異常的免疫反應(yīng)，主要用來治療器官移植抗排斥反應(yīng)和自身免疫性疾病。雖然近年來，新型免疫抑制劑不斷問世，但多數(shù)具有選擇性不高，可降低機(jī)體的正常免疫應(yīng)答、誘發(fā)感染和腫瘤等問題。 研究發(fā)現(xiàn)PI3K/Akt/mTOR信號(hào)通路在一些腫瘤及免疫性疾病中處于激活狀態(tài)。雖然已經(jīng)研發(fā)出專門針對(duì)單一蛋白激酶的免疫抑制劑。但是這些抑制劑在治療免疫疾病時(shí)仍然存在一些弊端。為了尋找新型選擇性強(qiáng)、作用快速、效果明顯、不良反應(yīng)少的免疫抑制劑，本課題在計(jì)算機(jī)輔助藥物設(shè)計(jì)（CADD）的基礎(chǔ)上，以PI3K蛋白激酶和mTOR蛋白為受體，篩選出先導(dǎo)化合物，，并運(yùn)用逆合成的方法，合成了一系列具有免疫抑制活性的含N雙雜環(huán)酰胺類化合物。為進(jìn)一步開發(fā)新型的PI3K/mTOR雙重免疫抑制劑提供了實(shí)驗(yàn)依據(jù)。 本課題研究的主要內(nèi)容： 1）先導(dǎo)化合物的確定 運(yùn)用Sybyl2.0分別以PI3K和mTOR作為受體，對(duì)ZINC,PubMed和BindingDB中下載到的近200萬個(gè)小分子進(jìn)行虛擬篩選。通過對(duì)虛篩得到的小分子進(jìn)行生物活性測(cè)試，確定先導(dǎo)化合物。 2）含N雙雜環(huán)酰胺類化合物的合成及純化 運(yùn)用逆合成的方法，分析合成路線，以最合理的合成方案合成一系列先導(dǎo)化合物的衍生物。通過液質(zhì)聯(lián)用技術(shù)以及波普技術(shù)，分析確定其結(jié)構(gòu)。 3）含N雙雜環(huán)酰胺類化合物對(duì)Jurkat T淋巴細(xì)胞的活性測(cè)定 基于CCK-8技術(shù)，測(cè)定所合成的化合物對(duì)于T淋巴細(xì)胞系的急性白血病細(xì)胞Jurkat的體外抑制活性。 通過上述研究，所得研究成果如下： 1）確定了3-(2,6-二氯苯基)-5-甲基-N-2-(6-甲磺酰苯并噻唑基)異惡唑-4-甲酰胺作為先導(dǎo)化合物。在此基礎(chǔ)上以2,6-二氯苯甲醛為起始原料經(jīng)過6步反應(yīng)，合成了包含先導(dǎo)化合物在內(nèi)的12個(gè)目標(biāo)化合物。經(jīng)高效液相（HPLC）檢測(cè)純度都達(dá)到98%以上，質(zhì)譜數(shù)據(jù)與核磁數(shù)據(jù)均與目標(biāo)化合物相符。 2）經(jīng)CCK-8法測(cè)試含N雙雜環(huán)酰胺類化合物對(duì)Jurkat細(xì)胞的增值抑制，獲得了有效可靠的數(shù)據(jù)。驗(yàn)證了化合物對(duì)于Jurkat T淋巴細(xì)胞的殺傷作用。 3)本課題所設(shè)計(jì)的含氮雙雜環(huán)酰胺類化合物具有良好的免疫抑制作用，為進(jìn)一步研究該類化合物，開發(fā)PI3K/mTOR雙重抑制劑提供了良好的實(shí)驗(yàn)基礎(chǔ)。
[Abstract]:Immunosuppressant, an immunosuppressant, is a class of substances that attenuate tissue damage by inhibiting cellular and humoral immune responses. Although new immunosuppressants have been developed in recent years, most of them have low selectivity, which can reduce the normal immune response, induce infection and tumor. Studies have found that PI3K/Akt/mTOR signaling pathways are activated in some tumors and immune diseases. Although immunosuppressants have been developed specifically for single protein kinases, these inhibitors are still present in the treatment of immune diseases. There are some disadvantages. In order to find a new type of selectivity, Immunosuppressants with rapid effect, obvious effect and less adverse reactions. Based on the computer aided drug design (CADDD), PI3K protein kinase and mTOR protein were used as receptors to screen the leading compounds, and the reverse synthesis method was used. A series of N-containing bicyclic amides with immunosuppressive activity were synthesized, which provide experimental basis for the further development of novel PI3K/mTOR double immunosuppressants. The main contents of this research are as follows:. 1) determination of lead compounds. PI3K and mTOR were used as receptors for virtual screening of nearly 2 million small molecules downloaded from BindingDB and pubMed by Sybyl2.0, and the leading compounds were determined by bioactivity test of the small molecules obtained by virtual screening. 2) Synthesis and purification of N-containing bicyclic amides. A series of derivatives of leading compounds were synthesized by the method of inverse synthesis, and their structures were determined by liquid-mass spectrometry and pop technique. 3) determination of the activity of N-containing bicyclic amides on Jurkat T lymphocytes. Based on CCK-8 technique, the inhibitory activity of the synthesized compounds on Jurkat of T lymphocyte line acute leukemia cells in vitro was determined. The results of the above studies are as follows:. 1) 3-trichlorobenzene 6-dichlorobenzene (2-Dichlorobenzyl) -5-methyl-N-2-thiazolyl) isoxazole-4-formamide was determined as the lead compound. On the basis of this, the starting material of 2C6-dichlorobenzaldehyde was used as the starting material, and the reaction was carried out in six steps. Twelve target compounds, including lead compounds, were synthesized. The purity of these compounds was over 98% by HPLC, and the mass spectrum data and the NMR data were all consistent with the target compounds. 2) the inhibitory effect of N-containing bicyclic amides on the proliferation of Jurkat cells was tested by CCK-8 method, and the effective and reliable data were obtained, and the killing effect of the compounds on Jurkat T lymphocytes was verified. 3) the nitrogen-containing bicyclic amides have good immunosuppressive effect, which provides a good experimental basis for the further study of these compounds and the development of PI3K/mTOR double inhibitors.
【學(xué)位授予單位】：重慶理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R914.5;R96

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 孫文竹;楊潔;;潛在藥靶的發(fā)現(xiàn)和驗(yàn)證[J];世界臨床藥物;2010年03期

2 朱倫;陳增良;;mTOR的結(jié)構(gòu)與功能[J];國際病理科學(xué)與臨床雜志;2006年01期

3 劉魏;何廣衛(wèi);吳強(qiáng);徐云根;;免疫抑制劑的研究新進(jìn)展[J];藥學(xué)進(jìn)展;2010年05期

4 成志勇;梁文同;底勝峰;潘];;PTEN信號(hào)轉(zhuǎn)導(dǎo)通路與腫瘤的多藥耐藥[J];中國腫瘤生物治療雜志;2009年04期

本文編號(hào)：1607247