小分子化合物YF-452抑制血管新生的功能及机理研究

发布时间：2018-06-29 22:01

本文选题：YF-452 + 血管新生　；参考：《华东师范大学》2014年硕士论文

【摘要】：血管新生是一个复杂的过程,包括内皮细胞的增殖、迁移、突破基底膜并形成三维管状网络,它是机体生长发育过程中必不可少的过程。同时,血管新生在疾病的发生发展中也发挥着重要的作用,尤其是肿瘤。新的血管生长进入肿瘤内部,为其提供必需的氧气和营养物质,从而为肿瘤的生长和转移提供了便利条件。因此,抑制血管新生被认为是抗肿瘤的重要疗法之一。由于肿瘤细胞基因组不稳定容易突变而产生耐药性,所以和传统的靶向于肿瘤细胞的化疗相比,抗血管新生治疗呈现出一定的优势。血管新生受多种刺激因子和抑制因子的严格调控。其中VEGF在众多促血管新生的生长因子和细胞因子中,对肿瘤血管新生的调控起着重要作用且在肿瘤组织中高表达。VEGF通过与内皮细胞表面的受体结合发挥其生物学功能,即VEGFR1(Flt-1),负责细胞的有丝分裂和化学应答；VEGFR2(KDR/Flk-1)负责内皮细胞的形态发生。VEGFR2的激活导致下游多种胞内信号分子的激活,包括Src家族激酶,FAK、ERK以及PI3K/AKT激酶等,它们分别调控血管渗透性,促进内皮细胞的增殖、迁移和分化,并维持细胞的存活。因此,VEGFR2和这些胞内信号分子被认为是抑制肿瘤血管新生的重要靶点。目前,临床上已有多种VEGFR2的抑制剂用于治疗肿瘤。但是,近年来临床结果显示,大多数这些抗癌药物呈现出明显的副反应且价格比较昂贵,所以寻找新的具有自主知识产权的小分子血管新生抑制剂作为抗癌备选药物具有重要意义。本研究中,我们通过对实验室小分子化合物库中的药物进行筛选,得到一个小分子化合物YF-452并分析其在体外对内皮细胞的抑制效果。同时在体内模型中也检测了YF-452的活性并对它抑制血管新生的分子机制进行了探索。YF-452在体外能够明显抑制内皮细胞的迁移、侵袭和管状网络的形成。半体内大鼠胸主动脉环模型和两个体内模型(鸡胚尿囊膜模型和小鼠角膜微囊袋模型)也显示出YF-452能够显著抑制新生微血管的形成。通过进一步的研究发现,YF-452能够抑制小鼠异种移植模型中肿瘤的生长及瘤内血管新生,和对照组相比,小鼠注射4Omg/kg/d YF-452对肿瘤生长的抑制率达到70%。通过对肿瘤组织CD31免疫荧光染色也表明YF-452具有抗血管新生功能。同时对各组小鼠的组织器官切片进行HE染色,并未发现有明显的病理学改变,各组小鼠的体重也未发生明显的变化,说明YF-452在抑制肿瘤生长和血管新生的过程中并未产生明显的毒性。Western Blot结果显示YF-452能够抑制VEGF诱导的VEGFR2的活性及下游蛋白激酶Src、FAK、ERK的激活。综上所述,我们找到了一个潜在的小分子化合物YF-452,能通过靶向VEGFR2的激活抑制血管新生,并将有可能成为肿瘤治疗的备选药物。此外,我们还建立了高脂饮食+STZ诱导的II型糖尿病小鼠模型,并在此基础上建立了糖尿病小鼠后肢缺血模型和伤口愈合模型,用于我课题组另外一名同学关于治疗糖尿病并发症新药研究,为药物疗效的评价奠定基础。
[Abstract]:Angiogenesis is a complex process, including the proliferation, migration, breakthrough of the basement membrane and the formation of a three-dimensional tubular network. It is an essential process in the growth and development of the body. At the same time, angiogenesis is also playing an important role in the development of the disease, especially the tumor. It provides the necessary oxygen and nutrients to facilitate the growth and metastasis of the tumor. Therefore, inhibition of angiogenesis is considered to be one of the most important therapies for anti-tumor. New treatment presents certain advantages. Angiogenesis is regulated by a variety of stimulating factors and inhibitory factors. Among them, VEGF plays an important role in the regulation of neovascularization of neovascularization and the high expression of.VEGF in tumor tissues by binding to receptors on the surface of endothelial cells. Its biological function, VEGFR1 (Flt-1), is responsible for the mitosis and chemical response of cells; VEGFR2 (KDR/Flk-1) is responsible for the activation of.VEGFR2 in the morphology of endothelial cells leading to the activation of a variety of intracellular signaling molecules downstream, including Src family kinase, FAK, ERK, and PI3K/ AKT kinases, which regulate vascular permeability and promote endothelial refinement, respectively. The proliferation, migration and differentiation of cells and the survival of the cells are maintained. Therefore, VEGFR2 and these intracellular signaling molecules are considered as an important target for inhibition of neovascularization. At present, a variety of VEGFR2 inhibitors have been used in the treatment of tumors. However, in recent years, clinical results show that most of these anticancer drugs present obvious side reactions. It is of great importance to find a new small molecular vascular neoplastic inhibitor with independent intellectual property as an anticancer drug. In this study, we screened a small molecule compound YF-452 by screening the drugs in the laboratory small molecule compound library and analyzed its effect on endothelial cells in vitro. At the same time, the activity of YF-452 was also detected in the body model and its molecular mechanism of inhibiting angiogenesis was explored..YF-452 could inhibit endothelial cell migration, invasion and formation of tubular network in vitro. The rat thoracic aorta ring model and two individual internal model (chicken embryo allantoic membrane model and mouse model in vivo) The corneal microcapsule model also showed that YF-452 could significantly inhibit the formation of neovascularization. Through further study, YF-452 could inhibit the growth of tumor and the angiogenesis in the xenotransplantation model in mice. Compared with the control group, the inhibition rate of 4Omg/kg/d YF-452 on tumor growth in mice was 70%. through the tumor group. CD31 immunofluorescence staining also showed that YF-452 had anti angiogenic function. At the same time, the tissues and organs of each group were stained with HE, and no obvious pathological changes were found. The weight of mice in each group did not change obviously. It showed that YF-452 did not produce obvious poison in the process of inhibiting tumor growth and angiogenesis. The results of sex.Western Blot show that YF-452 can inhibit the activity of VEGFR2 induced by VEGF and the activation of the downstream protein kinase Src, FAK, ERK. In summary, we have found a potential small molecule compound YF-452, which can inhibit angiogenesis by targeting VEGFR2, and will be a candidate for cancer treatment. In addition, we also A model of II induced diabetic mice induced by high fat diet +STZ was established. On this basis, the hind limb ischemia model and wound healing model of diabetic mice were established, which was used for the study of new drugs for diabetic complications in the other class of our group, laying a foundation for the evaluation of the efficacy of the drug.
【学位授予单位】：华东师范大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R96

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