本文選題：免疫抑制劑 + 分子對(duì)接��； 參考：《重慶理工大學(xué)》2014年碩士論文

【摘要】：JAK3酪氨酸蛋白激酶是一種僅存在于骨髓和淋巴細(xì)胞中的非受體酪氨酸激酶。JAK3可介導(dǎo)JAK3-STAT5信號(hào)通路調(diào)控細(xì)胞核內(nèi)DNA的轉(zhuǎn)錄，最終影響淋巴細(xì)胞基因表達(dá)，從而引起一系列的免疫缺陷病。目前已上市或處于臨床研究的JAK3抑制劑，均會(huì)因?yàn)橥瑫r(shí)抑制其他JAK亞型的激酶而存在一系列的副作用。 虛擬篩選是計(jì)算機(jī)輔助藥物設(shè)計(jì)中常用到的新藥研發(fā)的方法，通過虛擬篩選可以降低化合物的篩選數(shù)目，，較快擊中目標(biāo)，同時(shí)提高先導(dǎo)化合物的發(fā)現(xiàn)效率。分子對(duì)接為常用的篩選方法，對(duì)于靶點(diǎn)已知的蛋白質(zhì)，可對(duì)靶標(biāo)蛋白進(jìn)行活性位點(diǎn)定位，位點(diǎn)形狀提問，分子形狀匹配，打分函數(shù)評(píng)價(jià)四步來尋找符合條件的化合物�；谇捌诒緦�(shí)驗(yàn)室在喹唑啉化合物的篩選及效應(yīng)研究的方面取得了較好的效果。本文研究中以3LXK為靶標(biāo)，擴(kuò)大虛擬篩選范圍，對(duì)其進(jìn)行結(jié)構(gòu)改造，尋求活性更好的免疫抑制劑。 本課題研究的主要內(nèi)容： 1)新型免疫抑制劑的設(shè)計(jì) 本課題從Binding Database、Drugbank、NCI等數(shù)據(jù)庫共下載794473個(gè)小分子結(jié)構(gòu)，以SYBYLX1.3，Discovery Studio2.55軟件的相關(guān)模塊進(jìn)行篩選，分子對(duì)接和分子動(dòng)力學(xué)計(jì)算。并根據(jù)對(duì)接結(jié)果來指導(dǎo)分子的改造。 2)新型喹唑啉類化合物的合成 以2-氨基-苯甲酸類化合物、2-氰基-4'-甲基聯(lián)苯及丁胺為原料合成了4-{N-丁基-N-[(2'-1H-四唑-5-基]聯(lián)苯基)甲基]胺}-2-R1-6-R2-7-R3-喹唑啉化合物（R1=Me，R2=R3=H；R1=R3=H，R2=OCH3、OH；R1=H，R2=R3=OH，OCH3）。 3)新型喹唑啉類化合物的生物活性測(cè)定 采用CCK8檢測(cè)不同濃度的DMSO對(duì)Jurkat細(xì)胞增值的影響； 測(cè)定Jurkat細(xì)胞在96孔板內(nèi)的生長(zhǎng)曲線； CCK8檢測(cè)上述不同藥物濃度下的Jurkat細(xì)胞的影響。 取得的研究成果： 1)虛擬篩選確定了以4-{N-丁基-N-[(2'-1H-四唑-5-基]聯(lián)苯基)甲基]胺}-6-羥基喹唑啉作為先導(dǎo)化合物。 2)分子構(gòu)效關(guān)系研究設(shè)計(jì)出7個(gè)新型的喹唑啉化合物從而設(shè)計(jì)出了一系列的以4-{N-丁基-N-[(2'-1H-四唑-5-基]聯(lián)苯基)甲基]胺}-2-R1-6-R2-7-R3-喹唑啉化合物（R1=Me，R2=R3=H；R1=R3=H，R2=OCH3、OH；R1=H，R2=R3=OH，OCH3）。 3)通過LC-MS和1H NMR與13C NMR確證了純度和結(jié)構(gòu)； 4)在0.25%（V/V）下DMSO的濃度對(duì)Jurkat細(xì)胞增值并不會(huì)產(chǎn)生影響； 5)繪制了Jurkat細(xì)胞在96孔板內(nèi)的生長(zhǎng)曲線； 6)該類四唑喹唑啉化合物均對(duì)細(xì)胞增殖具有抑制作用，隨著喹唑啉類化合物濃度變大，細(xì)胞增值變緩，呈現(xiàn)一定的量-效關(guān)系。其中以N-{(2'-(2H-四氫吡唑-5-基)-[1,1'-聯(lián)苯]-4-基)甲基)-N-丁基-4-胺}-6，7-二羥基喹唑啉效果最好。結(jié)論：計(jì)算機(jī)輔助藥物設(shè)計(jì)在免疫抑制方面能指導(dǎo)新型免疫抑制劑的合成；本課題所設(shè)計(jì)的新型喹唑啉類化合物能有效地抑制Jurkat細(xì)胞增殖，并為尋求更好的JAK3抑制劑奠定基礎(chǔ)。
[Abstract]:JAK3 tyrosine protein kinase is a non-receptor tyrosine kinase. JAK3, which exists only in bone marrow and lymphocytes, can mediate JAK3-STAT5 signaling pathway to regulate the transcription of DNA in the nucleus and ultimately affect the gene expression of lymphocytes.This leads to a series of immune deficiency diseases.JAK3 inhibitors, which are currently listed or in clinical studies, have a series of side effects due to the simultaneous inhibition of other JAK subtypes of kinases.Virtual screening is a new drug research and development method commonly used in computer-aided drug design. Virtual screening can reduce the number of compounds, hit the target quickly, and improve the efficiency of the discovery of lead compounds.Molecular docking is a common screening method. For proteins with known targets, target proteins can be located at active sites, locus shape questioning, molecular shape matching and scoring function evaluation four steps to find suitable compounds.Good results were obtained in the screening and effect study of quinazoline compounds in our laboratory.In this study, 3LXK was used as a target to expand the scope of virtual screening, to reconstruct its structure, and to seek immunosuppressants with better activity.The main contents of this research are as follows:1) Design of novel immunosuppressantA total of 794473 small molecular structures were downloaded from the database of Binding Database / Drugbank / NCI. The molecular docking and molecular dynamics calculation were carried out by using the relative modules of SYBYLX1.3 Discovery Studio2.55 software.And according to the docking results to guide the modification of the molecule.2) Synthesis of novel quinazoline compoundsThe synthesis of 4- {N- Ding Ji -N- [2-(2 -) -1H-(-) -tetrazole-5-yl] methyl] amine} -2-R1-6-R2-7-R3-quinazoline compound R1MeR1-6-R7-R3-quinazoline compound R _ 1MeR _ 2H _ 3H _ 3H _ (1) (R _ 3H _ (1) H _ (1)) H _ (2) O _ (1) H _ ((1)) O _ ((1)) H _ (1) H _ (1) O _ (1) H _ (1) O _ ((1)) H _ (2) O _ ((1)) H _ ((1)) O _ ((1)) -azolidoline) was studied.3) determination of bioactivity of novel quinazoline compoundsCCK8 was used to detect the effect of different concentrations of DMSO on the proliferation of Jurkat cells.The growth curve of Jurkat cells in 96-well plate was measured.CCK8 was used to detect the effect of Jurkat cells at different drug concentrations.Research findings:1) using 4- {N- Ding Ji-N- [2H-1H-tetrazolyl]-methyl] amine}-6-hydroxyquinazoline as the lead compound.2) Seven new quinazoline compounds were designed by molecular structure-activity relationship, and a series of 4- {N- Ding Ji -N- [2 + -1H-tetrazole-5-yl] methyl] amine} -2-R1-6-R2-7-R3-quinazoline compound R1MeR2R3HN, R1R1OOOCH3OOOCH3OCH3OCH3OCH3OCH3.3) the purity and structure were confirmed by LC-MS, 1H NMR and 13C NMR.4) the concentration of DMSO had no effect on the proliferation of Jurkat cells at 0.25 / V;5) the growth curve of Jurkat cells in 96-well plate was plotted.6) all of these tetrazolium quinazoline compounds inhibited cell proliferation. With the concentration of quinazoline compounds increasing, cell proliferation slowed down, showing a dose-effect relationship.Among them, N- {N- {THP-2H-tetrahydropyrazole-5-[1- (1 +)-biphenyl]-4-yl) methyl-N--N--Ding Ji-4-amine}-6-7-dihydroxyquinazoline has the best effect.Conclusion: computer aided drug design can direct the synthesis of new immunosuppressants in the aspect of immunosuppression. The new quinazoline compounds designed in this paper can effectively inhibit the proliferation of Jurkat cells and lay a foundation for seeking better JAK3 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

【參考文獻(xiàn)】

中國(guó)期刊全文數(shù)據(jù)庫 前5條

1 羅小民,蔣華良,沈建華,陳凱先;藥物分子設(shè)計(jì)研究進(jìn)展[J];中國(guó)科學(xué)院院刊;2003年04期

2 陳燕,郝敬來,仇綴百;軟藥設(shè)計(jì)[J];中國(guó)臨床藥學(xué)雜志;2004年04期

3 文玉華,朱如曾,周富信,王崇愚;分子動(dòng)力學(xué)模擬的主要技術(shù)[J];力學(xué)進(jìn)展;2003年01期

4 陳瑋,黃象男;計(jì)算機(jī)輔助藥物設(shè)計(jì)研究[J];三門峽職業(yè)技術(shù)學(xué)院學(xué)報(bào);2003年01期

5 劉軍虎;劉勇;蹇軍友;鮑小平;;含1,2,4-三唑席夫堿的新型喹唑啉類化合物的合成及其抗菌活性研究[J];有機(jī)化學(xué);2013年02期

本文編號(hào)：1751505