新型肿瘤血管生成抑制剂来那度胺衍生物的设计与合成

发布时间：2018-05-10 04:40

  本文选题：肿瘤血管生成抑制剂 + 来那度胺衍生物　； 参考：《北京工业大学》2015年硕士论文

【摘要】：随着分子生物学研究的不断深入,恶性肿瘤的分子靶向治疗已成为除手术、放疗和化疗之外的第四种治疗方法,越来越多的用于临床治疗恶性肿瘤。血管生成与肿瘤的生长、侵袭和转移密切相关,以抑制肿瘤血管生成为靶点治疗肿瘤的方法已经成为抗肿瘤新疗法之一。来那度胺(Lenalidomide)是一种新型免疫调节剂,具有抗肿瘤作用活性,主要表现在抑制肿瘤血管生成作用。本文通过对来那度胺衍生物抑制肿瘤血管生成分子靶点的虚拟筛选研究,设计了一系列来那度胺衍生物的分子结构。在优化来那度胺合成工艺的基础上,合成并得到一系列来那度胺衍生物,并对其进行药理活性研究,为来那度胺衍生物抗肿瘤作用机制的研究及其作为分子靶向肿瘤血管生成抑制剂的研发提供理论与实验基础。首先,来那度胺抑制肿瘤血管生成分子靶点的筛选,及其抑制剂分子结构的设计和虚拟筛选。采用反向对接方法,以来那度胺(化学名为3-(4-氨基-1,3-二氢-1-氧代-2H-异吲哚-2-基)-2,6-哌啶二酮)为初始结构,在AutoDock 4.0平台上,选取六个血管生成促进因子靶蛋白,即血管内皮生长因子受体(VEGFR-2)、表皮生长因子受体(erbB-3)、成纤维细胞生长因子受体(FGFR-4)、BCR-ABL酪氨酸激酶(ABL)、p38丝裂原活化蛋白酶(p38MAPK)和基质金属蛋白酶(MMP-3)进行对接研究。通过分析对接结果,初步确定来那度胺抑制肿瘤血管生成的潜在靶点是FGFR-4。采用分子对接法,对设计的54个来那度胺衍生物与FGFR-4进行对接研究,确定拟合成包括烷基化衍生物、酰基化衍生物和磺酰化衍生物在内的30个目标化合物结构。其次,来那度胺及其衍生物的合成研究。通过对来那度胺及其中间体合成方法的综述,确定来那度胺的合成路线。以来那度胺为原料,通过对其氨基的衍生化研究(烷基化、酰基化和磺酰化反应),合成出30个来那度胺衍生物。通过考察溶剂,温度,催化剂,缩合剂和缚酸剂对合成过程中各步反应的影响,确定了各步反应的最佳反应条件。所合成的目标化合物均通过核磁共振氢谱、核磁共振碳谱和质谱等技术进行结构表征,其中28个化合物未见报道。最后,来那度胺及其衍生物的体外抗肿瘤活性研究。采用CCK-8法,进行来那度胺及其衍生物对食管癌细胞株EC9706的细胞毒性测试。结果表明,来那度胺及其衍生物均具有一定的抑制活性,来那度胺对EC9706的抑制活性IC50为340.3μg/m L(ΔG=-7.05 kcal/mol),而所选取的三个来那度胺衍生物对EC9706的抑制活性IC50分别为309.5μg/mL(ΔG=-7.19 kcal/mol),263.8μg/mL(ΔG=-7.68kcal/mol)和225.2μg/mL(ΔG=-8.40 kcal/mol),发现经过修饰的烷基化,酰基化和磺酰化来那度胺衍生物比来那度胺具有更好的抑制活性,其中磺酰化衍生物的抑制活性最为突出。此外,结果还表明来那度胺及其衍生物对EC9706的细胞毒性测试实验结果与分子对接结果相一致,说明将抑制肿瘤血管生成分子靶点作为来那度胺及其衍生物的研究方向是可行的,可进一步确定FGFR-4就是来那度胺抗肿瘤血管生成的作用靶点。
[Abstract]:With the continuous development of molecular biology research, the molecular targeting therapy of malignant tumor has become the fourth treatment methods except surgery, radiotherapy and chemotherapy, and more and more clinical treatment of malignant tumors. Angiogenesis is closely related to tumor growth, invasion and metastasis, in order to inhibit tumor angiogenesis as a target for tumor treatment. The method has become one of the new antitumor therapies. Lenalidomide is a new immunomodulator with antitumor activity, which is mainly manifested in inhibiting tumor angiogenesis. In this paper, a series of amines were designed by the virtual screening study on the molecular targets of tumor angiogenesis inhibition by the ramification. The molecular structure of the derivatives. Based on the optimization of the synthesis of amines, a series of amines were synthesized and obtained, and their pharmacological activities were studied. The research on the anti-tumor mechanism of the amines derivatives and the theoretical and experimental basis for the development of molecular targeting tumor angiogenesis inhibitors were provided. The screening of the molecular targets of tumor angiogenesis by amido, and the design and virtual screening of the molecular structure of the inhibitors. Using the reverse docking method, the initial structure of the amine (chemical named 3- (4- amino -1,3- two hydrogen -1- -2H- isoindole -2- based) -2,6- piperidine two ketone) was the initial structure, and six angiogenesis was selected on the AutoDock 4 platform. Promoting factor target proteins, such as vascular endothelial growth factor receptor (VEGFR-2), epidermal growth factor receptor (erbB-3), fibroblast growth factor receptor (FGFR-4), BCR-ABL tyrosine kinase (ABL), p38 mitogen activated protease (p38MAPK) and matrix metalloproteinase (MMP-3), are studied. By analyzing the docking results, the initial determination is made. The potential target of diamine inhibition of tumor angiogenesis is that FGFR-4. uses molecular docking to dock the designed 54 amines with FGFR-4 and determine the synthesis of 30 target compounds, including alkylated derivatives, acyl derivatives and sulfonylated derivatives. Secondly, the combination of amines and its derivatives A synthetic route to the synthesis of amines and their intermediates was reviewed to determine the synthetic route of the amines. Since the amine was used as a raw material, 30 diamine derivatives were synthesized by the study of its amino derivatives (alkylation, acylation and sulfonylation). Through the investigation of solvent, temperature, catalyst, condensation agent, and acid binding agent, The optimum reaction conditions for each step reaction were determined in the process of synthesis. The synthesized target compounds were characterized by nuclear magnetic resonance spectroscopy, nuclear magnetic resonance carbon spectroscopy and mass spectrometry, and 28 compounds were not reported. Finally, the antitumor activity of the amine and its derivatives in vitro was studied by CCK-8. The cytotoxicity test of the amines and their derivatives on the esophageal cancer cell line EC9706 showed that both amines and their derivatives had certain inhibitory activity, and the inhibitory activity of amines to EC9706 was 340.3 u g/m L (delta G=-7.05 kcal/mol), and the inhibitory activity of the three amino derivatives on EC9706 was selected. IC50 is 309.5 mu g/mL (delta G=-7.19 kcal/mol), 263.8 g/mL (delta G=-7.68kcal/mol) and 225.2 mu g/mL (delta G=-8.40 kcal/mol). It is found that the modified alkylation, acylation and sulfonylation of amines have better inhibitory activity than lauryl amine, and the inhibitory activity of sulfonylated derivatives is most prominent. The results of the cytotoxicity test of amidodiamine and its derivatives to EC9706 are in agreement with the molecular docking results, indicating that it is feasible to inhibit the molecular target of tumor angiogenesis as the direction of the amines and its derivatives. It can be further determined that FGFR-4 is the target of antitumor angiogenesis of amido.

【学位授予单位】：北京工业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R91;R914.5
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本文编号：1867828