在水介质中酶促合成细胞粘附位点RGD三肽前体及RGD聚乳酸微球的制备

发布时间：2019-04-30 13:15

【摘要】：细胞外基质中的粘附糖蛋白在细胞粘附中起重要作用,RGD三肽是许多粘附蛋白所共有的细胞间识别的最小序列,这一序列在介导细胞粘附、迁移和生长方面起重要作用。目前关于RGD的研究主要集中在RGD肽对血小板功能的影响、RGD肽在抗栓药物中的应用、RGD肽在骨组织的诱导再生中的研究、RGD肽在肿瘤转移和新生血管中的研究以及肿瘤靶向治疗中的应用。 在本文中,我们主要讨论了RGD三肽前体Bz-RGD-OMe的合成反应,这是个在动力学控制下的酶促肽合成反应。首先,合成Bz-RG-OEt,这是在胰蛋白酶催化、有机溶剂水共混体系下进行的,反应的最佳条件为:pH 8.0, 30℃,乙醇/ This-HCl缓冲液(85:15, V/V),酰基供体与亲核试剂的摩尔比为1:5,Bz-Arg-OEt 0.05mM。二肽的产率为70.6%。然后,合成Bz-RGD-OMe,这是个木瓜蛋白酶催化的可溶性三肽前体在水相介质中进行的合成反应。反应的最佳条件为:pH8.1,40℃,KCl水溶液反应体系(含有8mM EDTA和2mM DTT),底物Bz-RG(-OEt)和D(-OMe)2?HCl分别为0.05M和0.15M,反应时间为60min,最大转化率为62.41%。此外,还采用传统的液相化学法合成了游离RGD三肽。在论文的后部分,我们对RGD-聚乳酸微球的制备进行了初步探讨,并采用正交试验设计优化了试验条件,最后得到最优包封率为83.124%。
[Abstract]:Adhesion glycoproteins in extracellular matrix play an important role in cell adhesion. RGD tripeptide is the least recognized sequence common to many adhesion proteins, which plays an important role in mediating cell adhesion, migration and growth. At present, the research on RGD mainly focuses on the effect of RGD peptide on platelet function, the application of RGD peptide in antithrombotic drugs, and the study of RGD peptide in the induction and regeneration of bone tissue. The study of RGD peptide in tumor metastasis and neovascularization and the application of tumor targeting therapy. In this paper, we mainly discuss the synthesis of RGD tripeptide precursor Bz-RGD-OMe, which is an enzymatic peptide synthesis reaction controlled by kinetics. First, the synthesis of Bz-RG-OEt, was carried out in a trypsin-catalyzed, organic solvent-water blend system. The optimum reaction conditions were as follows: pH 8.0, 30 鈩，

本文编号：2468785