用于脑胶质瘤诊疗一体化的高分子递药系统研究
发布时间：2018-07-20 16:02
【摘要】：现代临床医疗手段逐渐从传统的"one-size-fits-all"向新型个体化治疗给药策略转变。这样的治疗方法使医生可以最大限度缩短诊断和治疗之间的时间。诊疗一体化治疗,就是把诊断试剂、治疗试剂整合在一个单独系统内,此种治疗策略在个体化给药领域起到重要的作用。最近诊断试剂普遍使用类型为:超声(US)、核磁共振(MR)、断层扫描(CT)、单光子发射计算机化断层显像(SPECT)等。通过发挥各自的优势,把诊断与治疗试剂组合在一起,起到协同作用,达到对实时监测治疗反馈的作用。同时,诊疗一体化纳米粒可以作为一种工具,用来筛选既安全又有效的剂量,使得医生在发病早期阶段避免药物不合理利用而导致对人体的副作用。在众多载体种类中,高分子是最受欢迎的纳米诊疗一体化平台。在癌症治疗过程中,作为载体平台系统,高分子聚合物可以通过化学键偶联方法,增加游离药物的溶解度和包封率,同时可以延长药物在体内的半衰期,保证药物在体内的释放。另外,高分子聚合物溶液中的粒径大小,通过增强渗透和保留(EPR)作用,实现被动靶向。而且除被动靶向之外,还可以在高分子链上连接主动靶向分子,准确靶向病灶部位,使得药物可以准确、高效运送到疾病部位。尽管上述以高分子为载体的诊疗一体化纳米粒发展迅猛、具有很多优点,但是现在仍然遇到很多问题需要迫切解决。第一,成本高。无毒安全的造影剂和高分子材料价格都很高,一方面,这样无形中增加了研发的成本,很多研发实验室并没有充足的资金去完成大批量的研究,另一方面,增加了病人的就医成本,很多病人并不能担负的起昂贵的高端药物费用,最后导致疾病进展快,耽误及时治疗;第二,在造影剂领域,尤其是涉及到造影剂自身功能与高分子载体相互作用时候物理化学性质方面的更深层次的研究很少,这对于诊疗一体化纳米粒子整体在机体的作用效果和安全性尤为重要。第三,制备方法繁琐,容易破坏药物本身功能基团,引入不必要的杂质,最终导致重复试验差,同时也不能大规模的生产。为了解决上述问题,本课题设计一种制备简单、安全性高、药效好的诊疗一体化纳米粒,对抗肿瘤递药系统有非常大的意义、为今后个体化用药领域深入研究有重要的参考价值。本课题的设计思路为,化学键偶联紫杉醇的聚谷氨酸高分子在水中可以自组装形成纳米粒,纳米粒外部溶于水、内部疏水,形成一个高分子载药平台。然后将溶于有机溶剂的造影剂逐滴加入水相中,通过超声乳化使其变成澄清的溶液,二者作用在统一系统内,再把多余溶剂去除,最终冷冻干燥形成诊疗一体化纳米粒子pGGA-PTX/SPION。通过动态蒸发光散射仪、透射电子显微镜、紫外分光光度计、热重分析仪等精密仪器,对此多功能纳米粒进行物理化学表征;用CCK-8试剂盒、普鲁士蓝染色、HE染色等对其抑制肿瘤效果以及对正常组织安全性进行系统的体外评价;动物体内实验包括两个方面,一方面通过核磁共振仪对此纳米粒的造影效果进行评价,另一方面是通过动物造模,尾静脉注射药物考察药效实验。通过上述一系列的实验,结果证明本实验制备的诊疗一体化纳米粒具有很好的诊断和治疗效果。在物理化学表征方面,pGGA-PTX/SPION纳米粒形态饱满均一,大小匀称,约110nm。通过EPR效应,被动靶向肿瘤组织。纳米粒载药量约为27.66%,造影剂含量约为6.34%,载药量高。在体外评价中,细胞存活率pGGA-PTX/SPION与PTX相似,说明有很好的抑制肿瘤细胞的作用,在HE实验中,此纳米粒对正常组织没有损害作用,充分体现了其良好的安全性能。在体内造影方面,通过核磁共振,我们可以清楚看到,药物在肿瘤体内整个代谢过程,实现了药物实时监测。体内药效实验中,pGGA-PTX/SPION组抑制肿瘤增长的效果最明显。综上所述,我们制备的诊疗一体化纳米粒子在未来的肿瘤诊断与治疗联合应用方面起到重要的作用,提供一种安全、有效、可行的科学研究思路。
[Abstract]:Modern clinical medical methods have gradually changed from the traditional "one-size-fits-all" to the new individualized treatment strategy. This treatment enables doctors to minimize the time between diagnosis and treatment. Integrated treatment is the integration of diagnostic reagents and therapeutic agents in a single system. Individualization plays an important role in the field of medicine. Recently, the types of diagnostic reagents are commonly used: ultrasound (US), nuclear magnetic resonance (MR), fault scan (CT), single photon emission computerized tomography (SPECT) and so on. By playing their respective advantages, the diagnosis and treatment test agents are combined together to achieve the feedback of real-time monitoring and treatment. At the same time, integrated nanoparticles can be used as a tool to screen both safe and effective doses so that doctors can avoid the irrational use of drugs at the early stage of the disease and lead to adverse effects on the human body. In many kinds of carrier species, polymer is the most popular platform for the integration of nano diagnosis and treatment. As a carrier platform system, high molecular polymer can increase the solubility and encapsulation efficiency of the free drug by chemical bond coupling method, at the same time, it can prolong the half-life of the drug in the body and ensure the release of the drug in the body. In addition, the size of the particle in the polymer solution can be achieved by enhancing the effect of EPR. Besides the passive target, besides the passive targeting, the active target molecules can be connected to the polymer chain, and the target location is accurately targeted. The drug can be accurately transported to the site of the disease. Although the integrated nanoparticles with high molecular weight have developed rapidly and have many advantages, there are still many problems to be encountered. The cost is high. On the one hand, it increases the cost of research and development, and many R & D laboratories do not have sufficient funds to complete large quantities of research. On the other hand, the cost of medical treatment is increased, and many patients can not afford to be expensive. The cost of high end drugs eventually leads to the rapid progress of the disease and delays the timely treatment. Second, in the field of contrast agents, in particular the deeper studies involving the physical and chemical properties of the contrast agent's own function and the interaction of the polymer carrier, this has a good effect on the diagnosis and treatment of the whole body in the body. Third, the preparation method is very complicated, easy to destroy the functional group of the drug itself, introduce unnecessary impurities, eventually lead to the poor repetition test, and can not be produced on a large scale at the same time. In order to solve the above problems, we design a simple, safe and effective medical treatment integrated nanoparticle, against the tumor delivery system. It has great significance for further research in the field of individualized drug use in the future. The design idea of this topic is that polyglutamic acid polymers with chemical bond coupling paclitaxel can form nanoparticles in water, and the nanoparticles are dissolved in water and internal hydrophobic, forming a polymer carrier platform. The contrast agent of the solvent is added to the water phase by drop by drop into a clarified solution by phacoemulsification. The two acts in a unified system, then the superfluous solvents are removed, and the final freeze drying is used to form an integrated nanoparticle pGGA-PTX/SPION. for diagnosis and treatment through a dynamic evaporative light scattering instrument, a transdermal electron microscope, a UV spectrophotometer, and a thermogravimetric analysis. A physical and chemical characterization of the multifunction nanoparticles was carried out by a precision instrument such as a CCK-8 kit, Prussian blue staining, and HE staining. The effects of the tumor and the safety of normal tissue were evaluated in vitro. In vivo experiments in animals included two aspects, and one side through MRI was used for the effect of the nanoparticles. On the other hand, the pGGA-PTX/ SPION nanoparticles have a good diagnosis and treatment effect. In the physical and chemical characterization, the morphology of the nanoparticles is full and uniform in size, about 110nm. The EPR effect was used to target the tumor tissue passively. The drug loading amount was about 27.66%, the contrast agent content was about 6.34%, and the drug loading was high. In the in vitro evaluation, the cell survival rate was similar to that of the PTX, indicating that the tumor cells had a good inhibitory effect on the tumor cells. In the HE experiment, the nanorgrain had no damage to normal tissues and fully embodied it. Good safety performance. In vivo imaging, through nuclear magnetic resonance, we can clearly see that the drug can monitor the whole metabolic process in the body of the tumor. In vivo pharmacodynamics experiment, the effect of the pGGA-PTX/SPION group on inhibiting the growth of tumor is most obvious. In summary, we have prepared the integrated nanoparticles in the future. The combination of tumor diagnosis and treatment plays an important role in providing a safe, effective and feasible scientific research train of thought.
【学位授予单位】：华东师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R943
本文编号：2134048
[Abstract]:Modern clinical medical methods have gradually changed from the traditional "one-size-fits-all" to the new individualized treatment strategy. This treatment enables doctors to minimize the time between diagnosis and treatment. Integrated treatment is the integration of diagnostic reagents and therapeutic agents in a single system. Individualization plays an important role in the field of medicine. Recently, the types of diagnostic reagents are commonly used: ultrasound (US), nuclear magnetic resonance (MR), fault scan (CT), single photon emission computerized tomography (SPECT) and so on. By playing their respective advantages, the diagnosis and treatment test agents are combined together to achieve the feedback of real-time monitoring and treatment. At the same time, integrated nanoparticles can be used as a tool to screen both safe and effective doses so that doctors can avoid the irrational use of drugs at the early stage of the disease and lead to adverse effects on the human body. In many kinds of carrier species, polymer is the most popular platform for the integration of nano diagnosis and treatment. As a carrier platform system, high molecular polymer can increase the solubility and encapsulation efficiency of the free drug by chemical bond coupling method, at the same time, it can prolong the half-life of the drug in the body and ensure the release of the drug in the body. In addition, the size of the particle in the polymer solution can be achieved by enhancing the effect of EPR. Besides the passive target, besides the passive targeting, the active target molecules can be connected to the polymer chain, and the target location is accurately targeted. The drug can be accurately transported to the site of the disease. Although the integrated nanoparticles with high molecular weight have developed rapidly and have many advantages, there are still many problems to be encountered. The cost is high. On the one hand, it increases the cost of research and development, and many R & D laboratories do not have sufficient funds to complete large quantities of research. On the other hand, the cost of medical treatment is increased, and many patients can not afford to be expensive. The cost of high end drugs eventually leads to the rapid progress of the disease and delays the timely treatment. Second, in the field of contrast agents, in particular the deeper studies involving the physical and chemical properties of the contrast agent's own function and the interaction of the polymer carrier, this has a good effect on the diagnosis and treatment of the whole body in the body. Third, the preparation method is very complicated, easy to destroy the functional group of the drug itself, introduce unnecessary impurities, eventually lead to the poor repetition test, and can not be produced on a large scale at the same time. In order to solve the above problems, we design a simple, safe and effective medical treatment integrated nanoparticle, against the tumor delivery system. It has great significance for further research in the field of individualized drug use in the future. The design idea of this topic is that polyglutamic acid polymers with chemical bond coupling paclitaxel can form nanoparticles in water, and the nanoparticles are dissolved in water and internal hydrophobic, forming a polymer carrier platform. The contrast agent of the solvent is added to the water phase by drop by drop into a clarified solution by phacoemulsification. The two acts in a unified system, then the superfluous solvents are removed, and the final freeze drying is used to form an integrated nanoparticle pGGA-PTX/SPION. for diagnosis and treatment through a dynamic evaporative light scattering instrument, a transdermal electron microscope, a UV spectrophotometer, and a thermogravimetric analysis. A physical and chemical characterization of the multifunction nanoparticles was carried out by a precision instrument such as a CCK-8 kit, Prussian blue staining, and HE staining. The effects of the tumor and the safety of normal tissue were evaluated in vitro. In vivo experiments in animals included two aspects, and one side through MRI was used for the effect of the nanoparticles. On the other hand, the pGGA-PTX/ SPION nanoparticles have a good diagnosis and treatment effect. In the physical and chemical characterization, the morphology of the nanoparticles is full and uniform in size, about 110nm. The EPR effect was used to target the tumor tissue passively. The drug loading amount was about 27.66%, the contrast agent content was about 6.34%, and the drug loading was high. In the in vitro evaluation, the cell survival rate was similar to that of the PTX, indicating that the tumor cells had a good inhibitory effect on the tumor cells. In the HE experiment, the nanorgrain had no damage to normal tissues and fully embodied it. Good safety performance. In vivo imaging, through nuclear magnetic resonance, we can clearly see that the drug can monitor the whole metabolic process in the body of the tumor. In vivo pharmacodynamics experiment, the effect of the pGGA-PTX/SPION group on inhibiting the growth of tumor is most obvious. In summary, we have prepared the integrated nanoparticles in the future. The combination of tumor diagnosis and treatment plays an important role in providing a safe, effective and feasible scientific research train of thought.
【学位授予单位】：华东师范大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R943
【参考文献】
相关期刊论文 前1条
1 郑明彬;赵鹏飞;罗震宇;龚萍;张鹏飞;盛宗海;高冠慧;蔡林涛;;纳米技术在癌症诊疗一体化中的应用[J];科学通报;2014年31期
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