泛素化降解雌激素受体α的小分子化合物筛选及其初步抗肿瘤活性研究

发布时间：2018-04-22 17:17

本文选题：乳腺癌 + 雌激素受体α　；参考：《浙江省医学科学院》2017年硕士论文

【摘要】：[背景]:乳腺癌是女性常见肿瘤,近年来发病率显著增高。约86%的乳腺癌是激素依赖性疾病,内分泌治疗是乳腺癌重要的治疗手段。他莫昔芬是目前临床上使用最广泛有效的选择性雌激素受体调节剂,但其耐药性是临床应用后期的瓶颈。蛋白降解靶向嵌合体技术是将目标蛋白和细胞内的泛素连接酶结合,从而导致目标蛋白泛素化降解。嵌合体包括三部分:一头是靶向目标蛋白的小分子化合物结构;另一头是可以招募蛋白降解体系比如E3 Ligase的结构;中间通过合适的linker连接。利用蛋白降解靶向嵌合体技术为克服靶向治疗药物临床应用耐药性问题提供了新的治疗策略。[目的]:筛选出高结合力、高选择性的雌激素受体α的配体小分子化合物结构,进行初步抗肿瘤活性研究,为进一步合成可以同时抑制和靶向泛素化降解雌激素受体双重药效机制的嵌合体药物奠定基础。[方法]:以荧光偏振技术检测候选小分子化合物与雌激素受体α及β亚型的结合力及选择性,以双荧光素酶报告基因检测系统检测候选小分子化合物对雌激素受体α转录活性的影响,以免疫组织化学法以及Western Blot鉴定乳腺癌MCF-7细胞雌激素受体α的表达,以WST-1法检测候选小分子化合物对MCF-7细胞体外增殖的影响,以乳腺癌细胞裸小鼠皮下移植瘤模型观察候选小分子化合物对乳腺癌肿瘤体内生长的影响。[结果]:候选小分子化合物中4g和5g与雌激素受体α和β有较强结合力,其中化合物4g对雌激素受体α亚型的选择性强于他莫昔芬;双荧光素酶报告基因检测系统检测结果表明小分子化合物4g对雌激素受体α具有拮抗作用,对雌激素受体α表达阳性的人乳腺癌MCF-7细胞具有显著的增殖抑制作用,对接种MCF-7细胞的裸小鼠体内移植瘤具有显著的生长抑制作用。[结论]:候选小分子化合物4g对雌激素受体α具有较好的结合力和选择性,同时对乳腺癌具有抗肿瘤活性,是较为理想的构建具有同时抑制和靶向泛素化降解雌激素受体α嵌合体的小分子化合物结构。
[Abstract]:Background: breast cancer is a common tumor in women. The incidence of breast cancer has increased significantly in recent years. About 86% of breast cancer is hormone-dependent disease, endocrine therapy is an important treatment for breast cancer. Tamoxifen is the most widely used selective estrogen receptor modulator in clinical practice, but its resistance is the bottleneck in the later stage of clinical application. Protein degradation targeting chimera technology is to combine the target protein with the intracellular ubiquitin ligase, which leads to the degradation of the target protein ubiquitin. The chimera consists of three parts: one is the structure of small molecule which targets the target protein; the other is the structure of protein degradation system such as E3 Ligase; the middle is connected by suitable linker. The use of protein-degrading targeted chimerism provides a new therapeutic strategy for overcoming drug resistance in clinical application of targeted drugs. [objective]: to screen small ligand structures of estrogen receptor 伪 with high binding ability and selectivity, and to study its antitumor activity. It provides a basis for the further synthesis of chimeric drugs which can simultaneously inhibit and target the degradation of estrogen receptor by Ubiquitin. [methods]: the binding force and selectivity of candidate small molecular compounds with estrogen receptor 伪 and 尾 subtypes were detected by fluorescence polarization technique. Double luciferase reporter gene detection system was used to detect the effect of candidate small molecule compounds on the transcriptional activity of estrogen receptor 伪, and the expression of estrogen receptor 伪 in breast cancer MCF-7 cells was identified by immunohistochemical method and Western Blot. The effect of candidate small molecule compounds on the proliferation of MCF-7 cells in vitro was detected by WST-1 assay, and the effect of candidate small molecule compounds on the growth of breast cancer tumors in vivo was observed by subcutaneous transplantation of breast cancer cells in nude mice. [results]: among candidate small molecular compounds, 4 g and 5 g had strong binding ability with estrogen receptor 伪 and 尾, and the selectivity of compound 4 g to estrogen receptor 伪 subtype was stronger than that of tamoxifen. The results of double luciferase reporter gene detection system showed that small molecular compound 4 g had antagonistic effect on estrogen receptor 伪 and significantly inhibited the proliferation of human breast cancer MCF-7 cells with positive expression of estrogen receptor 伪. It can inhibit the growth of xenografts in nude mice inoculated with MCF-7 cells. [conclusion]: candidate small molecule compound 4 g has good binding ability and selectivity to estrogen receptor 伪, and has antitumor activity to breast cancer. It is an ideal structure of small molecular compounds with inhibition and targeting of Ubiquitin degradation of estrogen receptor 伪 chimerism.
【学位授予单位】：浙江省医学科学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R979.1

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