雷公藤甲素仿生纳米粒的构建及其用于黑色素瘤的治疗

雷公藤甲素仿生纳米粒的构建及其用于黑色素瘤的治疗目录目录.21摘要.42引言……62.1研究目的及意义。.62.2研究进展…72.2.1雷公藤甲素抗肿瘤的研究进展72.2.2构建纳米给药系统的研究进展.83方法.,83.1TPL@TFBF的制备83.1.1单因素考察3.1.21FBF的制备.103.1.3TPL@T℉的制备113.1.4TPL@TFB的制备.....113.1.5TPL@T℉BF的制备.113.2TPL@TFBF的表征与质量评价.113.2.1粒径和电位表征113.2.2外观及微观形态113.2.3紫外光谱分析.……113.3TPL@TFBF的体外稳定性..123.3.1胶体隐定性，，123.3.2储存稳定性…123.4TPL、TPL@TFBF和TPL@TFBF对B16F10细胞的细胞毒性...124结论..124.1TPL@TFBF的制备与表征.……124.2TPL@T℉BF的表征..174.3细胞毒性194.3.1TPL、TPL@TFBF和TPL@TFBF对B16F10的细胞毒性研究....194.4结论,205参考文献,216致谢.2311摘要雷公藤甲素(triptolide,TPL)是雷公藤这一天然药物的关键活性成分之一，其具有显著的抗炎及免疫抑制等药理作用，对多种疾病的治疗具有重要意义，此外还具有显著的抗肿瘤效果。然而，因其自身固有水溶性不佳、副作用较大以及毒性较强等问题，使其临床应用受到了极大的限制。为了解决这一限制，我们构建了一个纳米给药系统，用以克服雷公藤甲素在临床应用时所存在的问题。本文采用高效液相色谱法来建立TPL含量测定的方法并分别进行专属性实验、精密度实验、建立标准曲线以及进行加样回收率实验等方法学考察。采用单因素法考察BSA投药浓度，TPL投药浓度及TPL投药浓度对TPL载药量和包封率的影响，确定制备TPL@TFBF的最佳工艺，并对该优选条件下制备的纳米粒进行表征与质量评价，再考察其体外稳定性，最后采用CCK8法检测各制剂对B16F10细胞的细胞毒性。最终实验结果表明该纳米系统可以鞣酸(TA)为配体形成TA-F3+MOFs,用BSA-FA修饰的MOFs包被疏水药物TPL。这种含铁MOF和BSA-FA的结合增强了铁的内吞作用和靶向药物递送的效率，并抑制肿瘤细胞活性，提高雷公藤甲素的治疗效果。关键词：雷公藤甲素，黑色素瘤，金属有机骨架，肿瘤AbstractTriptolide (TPL)is one of the key active ingredients of Tripterygium wilfordii Hook.f.,anatural medicine.It has significant pharmacological effects such as anti-inflammatory andimmunosuppressive activities,and is of great significance for the treatment of various diseases.In addition,it also has remarkable anti-tumor effects.However,its poor water solubility,significant adverse reactions,and high toxicity have seriously limited its clinical application.Toaddress this limitation,we have constructed a nano-drug delivery system to overcome theproblems encountered in the clinical application of triptolide.In this study,we usedhigh-performance liquid chromatography to establish a method for determining the content oftriptolide (TPL)and conducted methodological investigations including specificity experiments,precision experiments,establishment of standard curves,and sample recovery experiments.Weused the single-factor method to investigate the effects of BSA concentration,TPL2concentration,and TPL concentration on the drug loading and encapsulation efficiency of TPL,in order to determine the optimal process for preparing TPL@TFBF.We then characterized andevaluated the quality of the nanoparticles prepared under the optimized conditions,examinedtheir stability in vitro,and finally used the CCK8 method to detect the cytotoxicity of eachpreparation against B16F10 cells.The final experimental results indicate that the nano-systemcan form TA-Fe3+MOFs with tannic acid (TA)as the ligand,and the MOFs modified byBSA-FA coats the hydrophobic drug TPL.The combination of iron-containing MOF andBSA-FA enhances the endocytosis of iron and the efficiency of targeted drug delivery,inhibitsthe activity of tumor cells,and improves the therapeutic effect of triptolide.Key Words:Triptolide,melanoma,metal-organic framework,tumor3