新型喹唑啉化合物的设计、合成及免疫抑制活性的初步研究

发布时间：2018-04-14 23:26

  本文选题：免疫抑制剂 + 分子对接　； 参考：《重庆理工大学》2014年硕士论文

【摘要】：JAK3酪氨酸蛋白激酶是一种仅存在于骨髓和淋巴细胞中的非受体酪氨酸激酶。JAK3可介导JAK3-STAT5信号通路调控细胞核内DNA的转录，最终影响淋巴细胞基因表达，从而引起一系列的免疫缺陷病。目前已上市或处于临床研究的JAK3抑制剂，均会因为同时抑制其他JAK亚型的激酶而存在一系列的副作用。 虚拟筛选是计算机辅助药物设计中常用到的新药研发的方法，通过虚拟筛选可以降低化合物的筛选数目，，较快击中目标，同时提高先导化合物的发现效率。分子对接为常用的筛选方法，对于靶点已知的蛋白质，可对靶标蛋白进行活性位点定位，位点形状提问，分子形状匹配，打分函数评价四步来寻找符合条件的化合物。基于前期本实验室在喹唑啉化合物的筛选及效应研究的方面取得了较好的效果。本文研究中以3LXK为靶标，扩大虚拟筛选范围，对其进行结构改造，寻求活性更好的免疫抑制剂。 本课题研究的主要内容： 1)新型免疫抑制剂的设计 本课题从Binding Database、Drugbank、NCI等数据库共下载794473个小分子结构，以SYBYLX1.3，Discovery Studio2.55软件的相关模块进行筛选，分子对接和分子动力学计算。并根据对接结果来指导分子的改造。 2)新型喹唑啉类化合物的合成 以2-氨基-苯甲酸类化合物、2-氰基-4'-甲基联苯及丁胺为原料合成了4-{N-丁基-N-[(2'-1H-四唑-5-基]联苯基)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)新型喹唑啉类化合物的生物活性测定 采用CCK8检测不同浓度的DMSO对Jurkat细胞增值的影响； 测定Jurkat细胞在96孔板内的生长曲线； CCK8检测上述不同药物浓度下的Jurkat细胞的影响。 取得的研究成果： 1)虚拟筛选确定了以4-{N-丁基-N-[(2'-1H-四唑-5-基]联苯基)甲基]胺}-6-羟基喹唑啉作为先导化合物。 2)分子构效关系研究设计出7个新型的喹唑啉化合物从而设计出了一系列的以4-{N-丁基-N-[(2'-1H-四唑-5-基]联苯基)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)通过LC-MS和1H NMR与13C NMR确证了纯度和结构； 4)在0.25%（V/V）下DMSO的浓度对Jurkat细胞增值并不会产生影响； 5)绘制了Jurkat细胞在96孔板内的生长曲线； 6)该类四唑喹唑啉化合物均对细胞增殖具有抑制作用，随着喹唑啉类化合物浓度变大，细胞增值变缓，呈现一定的量-效关系。其中以N-{(2'-(2H-四氢吡唑-5-基)-[1,1'-联苯]-4-基)甲基)-N-丁基-4-胺}-6，7-二羟基喹唑啉效果最好。结论：计算机辅助药物设计在免疫抑制方面能指导新型免疫抑制剂的合成；本课题所设计的新型喹唑啉类化合物能有效地抑制Jurkat细胞增殖，并为寻求更好的JAK3抑制剂奠定基础。
[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.
【学位授予单位】：重庆理工大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R914

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