多胺修饰环糊精—雷公藤活性化合物的超分子体系构建及其与BSA相互作用研究

发布时间：2018-10-23 20:12

【摘要】：雷公藤红素(CSL)、雷公藤甲素(TPL)和雷公藤Qg酯酮(Tri)均属于卫矛科雷公藤,是雷公藤主要活性成分。研究表明这些化合物具有抗氧化,抗类风湿,抗老年性痴呆症等功效,其显著的抗肿瘤活性方面已成为目前研究热点。但是其水溶性差、生物利用度低,并有较大的毒性,限制其进一步开发和应用。多胺修饰β-环糊精(CD)的特殊结构及两亲性质,与药物分子进行包合,“N”原子可增加识别位点,提高对药物分子的键合能力,从而显著提高药物分子的水溶性,使药物具有更好的应用前景。本文合成了4种多胺修饰β-CDs(H1-4),制备其与3种雷公藤活性化合物的12种超分子体系,并表征,对其可能的包结模式进行推测。采用紫外-吸收光谱法和荧光光谱法对H1-4/雷公藤活性化合物与牛血清白蛋白(BSA)的相互作用进行了研究。考察了H1/CSL的抗肿瘤活性,并进一步采用理论计算模拟研究H2/CSL和CSL与BSA的作用机制。本文的主要工作如下:1.本文合成了4种多胺修饰β-CDs(H1-4),并通过1H NMR、MS、IR等方法表征。2.制备H1-4与雷公藤活性化合物的12种超分子体系,通过XRD、1H NMR和IR等方法进行表征。H1-4/雷公藤活性化合物的可能包结模式根据2D ROESY谱图推测。紫外光谱实验表明,H1-4与雷公藤活性化合物均形成了1:1型的超分子体系,H1-4与雷公藤活性化合物之间的稳定常数大小顺序为KH4/CSLKH2/CSLKH3/CSLKH1/CSL、KH4/TPLKH2/TPLKH3/TPLKH1/TPL和KH4/TriKH2/TriKH3/TriKH1/Tri。水溶性实验表明,CSL、TPL、Tri与H1-4形成超分子体系后,水溶性分别最大提高63.5、57.6、137.3倍。3.采用紫外光谱法和荧光光谱法对H1-4/雷公藤活性化合物与BSA的相互作用进行了研究,表明H1-4/雷公藤活性化合物与BSA产生了静态猝灭,且以1:1形成复合物,结合过程的作用力为范德华和氢键力。同步荧光光谱法研究表明H1-4/雷公藤活性化合物的加入对BSA酪氨酸残基部位的结构产生了影响,但对其色氨酸残基部位均无影响,结合距离r均小于7 nm,表明偶极-偶极非辐射能量转移也可以引起BSA荧光猝灭。4.研究CSL和H1/CSL超分子体系与五株细胞肝癌SMMC-7721、结肠癌SW480、肺癌A-549、白血病HL-60和乳腺癌MCF-7细胞的体外抗癌活性及对人正常肺上皮细胞BEAS-2B的毒性。结果表明:CSL和H1/CSL超分子体系对五株细胞的体外抗肿瘤活性均高于对照物顺铂。形成H1/CSL超分子体系后,CSL的毒性有所降低。5.采用分子对接和分子动力学模拟研究了H2/CSL和CSL/BSA之间的作用机制,结果表明理论计算模拟结果与实验结果一致。
[Abstract]:Tripterygium wilfordii (CSL), triptolide (TPL) and Qg triptolide (Tri) belong to Tripterygium wilfordii and are the main active components of Tripterygium wilfordii. It has been shown that these compounds have anti-oxidation, anti-rheumatoid and anti-Alzheimer 's effects, and their significant antitumor activity has become a hot research topic at present. However, its poor water solubility, low bioavailability and toxicity limit its further development and application. The special structure and amphiphilic properties of 尾 -cyclodextrin (CD) modified by polyamines can be encapsulated with the drug molecules. "N" atoms can increase the recognition sites and the binding ability of the drug molecules, thus significantly improving the water-solubility of the drug molecules. So that the drug has a better application prospects. In this paper, four polyamines modified 尾-CDs (H1-4) were synthesized, 12 supramolecular systems were prepared and characterized with 3 active compounds of Tripterygium wilfordii. The interaction between H1-4 / Tripterygium wilfordii active compound and bovine serum albumin (BSA) (BSA) was studied by UV-absorption spectrometry and fluorescence spectrometry. The antitumor activity of H1/CSL was investigated, and the mechanism of H2/CSL, CSL and BSA was studied by theoretical simulation. The main work of this paper is as follows: 1. Four polyamines modified 尾-CDs (H1-4) were synthesized and characterized by 1H NMR,MS,IR. Twelve supramolecular systems of H1-4 and tripterygium wilfordii active compounds were prepared and characterized by XRD,1H NMR and IR. The possible inclusion patterns of H1-4 / Tripterygium wilfordii active compounds were deduced from 2D ROESY spectra. UV spectra showed that both H1-4 and tripterygium wilfordii active compounds formed 1:1 supramolecular system, and the order of stability constants between H1-4 and tripterygium wilfordii was KH4/CSLKH2/CSLKH3/CSLKH1/CSL,KH4/TPLKH2/TPLKH3/TPLKH1/TPL and KH4/TriKH2/TriKH3/TriKH1/Tri.. The water-solubility experiment showed that the water solubility of CSL,TPL,Tri and H1-4 was increased by 57.6137.3 times respectively after the formation of supramolecular system. The interaction between H1-4 / tripterygium wilfordii active compounds and BSA was studied by UV and fluorescence spectra. It was found that H1-4 / tripterygium wilfordii active compounds produced static quenching with BSA and formed complexes at 1:1. The binding forces are van der Waals and hydrogen bonding forces. Synchronous fluorescence spectroscopy showed that the addition of H1-4 / Tripterygium wilfordii had an effect on the structure of BSA tyrosine residues, but had no effect on the tryptophan residues. The binding distance r is less than 7 nm, which indicates that dipole-dipole non-radiation energy transfer can also cause fluorescence quenching of BSA. To study the anticancer activity of CSL and H1/CSL supramolecular system and five cell lines of SMMC-7721, colon cancer, SW480, lung cancer A-549, leukemia HL-60 and breast cancer MCF-7 cells in vitro and the toxicity to BEAS-2B of human normal lung epithelial cells. The results showed that the antitumor activity of CSL and H1/CSL supramolecular system was higher than that of cisplatin in vitro. After the formation of H1/CSL supramolecular system, the toxicity of CSL was decreased by 0.5. 5%. The interaction mechanism between H2/CSL and CSL/BSA is studied by molecular docking and molecular dynamics simulation. The results show that the theoretical simulation results are consistent with the experimental results.
【学位授予单位】：云南师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O643.1

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