生物大分子纳米载药体系的功能化设计及其在肿瘤诊疗中的研究
发布时间：2018-10-05 08:28
【摘要】：随着纳米科学技术的发展,纳米药物递送体系的应用为肿瘤的诊断和治疗提供了有效解决途径。纳米载药体系可将药物和成像试剂集成于纳米颗粒中,通过增强型渗透效应或主动靶向作用有针对性地递送到肿瘤组织,提高治疗效果和减少对正常组织的毒副作用。在众多纳米载药体系中,由于生物大分子纳米载药体系具备良好的生物相容性与生物可降解性,因此可避免其在体内长期累积所导致的毒性。基于生物可降解性纳米材料的优点,本论文围绕改善肿瘤诊疗效果这一目标,开发了基于牛血清白蛋白、藻蓝蛋白和DNA的生物大分子纳米载药体系,将其应用于肿瘤的靶向治疗、影像示踪、扭转耐药和放疗/化疗协同治疗等。主要的研究工作如下:第一章介绍了纳米药物在肿瘤诊疗中的发展现状,概述了生物大分子纳米载体的类别、优势及其在肿瘤诊断和治疗中的应用。在第二章中,为了提高放疗效果,合成了叶酸靶向的白蛋白纳米载药体系,并负载有机硒衍生物PSeD,构建了兼具肿瘤靶向性与生物相容性的白蛋白纳米载药体系(FA-BSANP@Se),其能联合X射线能大幅度提高宫颈癌Hela细胞内的活性氧水平,使其发生大规模DNA损伤,从而级联激活G2/M周期阻滞和凋亡信号,最终抑制Hela细胞克隆群落的形成。为了克服异源性BSANPs可能对人体造成的免疫源性,在第三章中制备了无免疫源性的新型蛋白纳米载药体系—藻蓝蛋白纳米粒子(Phycocyanin nanoparticles,PCNPs)作为阿霉素(doxorubicine,DOX)的新型纳米载体,并装载靶向分子叶酸(folate,FA),探究其在扭转肿瘤细胞对阿霉素耐药的作用及相关机制。药代动力学研究表明,PCNPs明显延长了DOX的血液半衰期,在酸性溶酶体环境中释放DOX。FA-PCNP@DOX通过FA/FARs介导的靶向作用识别肿瘤耐药细胞RHep G2,通过上调胞内活性氧水平引起线粒体断裂,继而阻断ATP依赖性ABC转运蛋白的活性,显著提高了DOX在RHepG2细胞内的吸收和停留效率,最终引发RHepG2细胞凋亡。活体成像实验证明,FA-PCNP@DOX能靶向肿瘤区域,在有效抑制瘤体增殖的同时降低DOX引起的肝、肺、肾毒性。进一步地,为了改善金属配合物的血液相容性和靶向抗肿瘤效果,在第四章中构建了cRGD靶向藻蓝蛋白纳米粒子作为钌多吡啶配合物(Ruthenium polypyridyl complexes,RuPOP)的载体,发现cRGD-PCNP@Ru能提高RuPOP的亲水性,并阻止RuPOP和血红细胞相互作用,从而改善了RuPOP的血液相容性。通过对一系列宫颈癌细胞Caski、Siha和Hela细胞进行活性筛选,发现cRGD-PCNP@Ru能通过RGD-整合素介导的靶向作用识别整合素高表达的宫颈癌细胞Caski,提高了RuPOP在Caski肿瘤球内部的累积,从而提高了RuPOP对Caski肿瘤球增殖的抑制作用。为了提高纳米载药体系和对金属配合物的负载效率及体内抗肿瘤活性,在第五章中构建了Biotin靶向DNA cages纳米载药体系作为金属配合物RuPOP的载体(Bio-Cage@Ru),研究其在提高RuPOP抗肿瘤活性和降低毒性方面的潜能。Bio-cage作为具有肿瘤靶向性的四面体笼型结构,通过生物素/受体介导的靶向作用增强了RuPOP对不同肿瘤细胞和正常细胞选择性,实现了良好的抗肿瘤效果。Bio-cage@Ru加速RuPOP定位至细胞核,并在DNase I的作用下裂解DNA结构,促进RuPOP的释放。在荷瘤裸鼠模型中,Bio-cage@Ru提高了RuPOP在肿瘤区域的累积,从而明显提高了其体内抗肿瘤效果。此外,Bio-cage@Ru明显降低了RuPOP所引起的心、肝、肺、肾毒性,整体实现了高效、低毒的目标。在第五章中对全文工作进行了总结,并对未来的研究方向提出了展望。本论文工作的开展为癌症治疗中出现的放疗、化疗效果差、毒副作用明显以及肿瘤耐药等问题提供了解决方法。
[Abstract]:With the development of nano science and technology, the application of nano drug delivery system provides an effective way for diagnosis and treatment of tumor. The nano drug loading system can integrate the drug and the imaging agent into the nano particles, and is targeted to the tumor tissue through the enhanced osmotic effect or the active targeting effect, thereby improving the treatment effect and reducing the toxic and side effects on the normal tissues. In many nano-drug delivery systems, since the biological macromolecule nano-drug carrier system has good biocompatibility and biodegradability, toxicity caused by long-term accumulation in vivo can be avoided. Based on the advantages of biodegradable nano materials, this paper aims at improving the diagnosis and treatment effect of tumor, and develops a biological macromolecule nano medicine loading system based on bovine serum albumin, phycoerythrin and DNA, which is applied to targeted therapy and image tracing of tumors. torsion resistance and radiotherapy/ chemotherapy co-treatment, etc. The main research work is as follows: The first chapter introduces the development status of nano-drugs in tumor diagnosis and treatment, and summarizes the categories, advantages and applications of biological macromolecule nano-carriers in tumor diagnosis and treatment. In the second chapter, in order to improve the effect of radiotherapy, folic acid-targeted albumin nano-drug delivery system was synthesized and the organic selenium derivative PSeD was loaded, and the albumin nano-drug delivery system (FA-BSANP@Se) with tumor targeting and biocompatibility was constructed. The combined X-ray can greatly improve the active oxygen level in the uterine cervix cancer cell and cause large-scale DNA damage, so that the G2/ M cycle block and the apoptosis signal are cascade activated, and finally the formation of the cell clone community of the cervical cancer cell is inhibited. In order to overcome the possible immune source of heterogenic BSANPs to human body, a new type of nano-carrier of phycoerythrin nanoparticles (PCNPs) as adriamycin (DOX) was prepared in the third chapter, and targeted molecular folic acid (Folate) was loaded. FA) to explore its role in reversing adriamycin resistance in tumor cells and its related mechanisms. Pharmacokinetic studies indicate that PCNPs significantly prolong the blood half-life of DOX, release DOX.FA-PCNP@DOX in acidic lysosomal environments to recognize tumor-resistant cells RHep G2 through FA/ FARs-mediated targeting, and cause mitochondrial breakage by up-regulation of intracellular reactive oxygen levels, and then blocking the activity of the ATP-dependent ABC transporter, remarkably improving the absorption and staying efficiency of DOX in the RHepG2 cells, and finally inducing the apoptosis of the RHepG2 cells. In vivo imaging experiments prove that the FA-PCNP@DOX can target tumor regions and reduce the toxicity of liver, lung and kidney caused by DOX while effectively inhibiting the proliferation of tumor bodies. Further, in order to improve the blood compatibility and targeted anti-tumor effect of metal complexes, we constructed the vector of cGVHD-targeting phycoerythrin nanoparticles as the carrier of polychrome complex (RuPOP) in the fourth chapter, and found that the cRGD-PCNP@Ru can improve the hydrophilicity of RuPOP. and prevents the interaction of RuPOP and red blood cells, thereby improving the blood compatibility of RuPOP. By carrying out active screening on a series of cervical cancer cells Caski, Siha, and Caski cells, it is found that the cRGD-PCNP@Ru can recognize the high-expression cervical cancer cell Caski through the targeting action mediated by the caspase-whole, and the accumulation of the RuPOP in the Caski tumor ball is improved, thereby improving the inhibiting effect of RuPOP on the proliferation of Caski tumor balls. In order to improve the loading efficiency and anti-tumor activity of the nano-drug delivery system and the metal complex, Biotin-targeted DNA images nano-drug delivery system was constructed as the carrier (Bio-Cage@Ru) of the metal complex RuPOP in chapter V to study its potential in improving the antitumor activity of RuPOP and reducing toxicity. Bio-Cage, as a tetrahedral cage type structure with tumor targeting property, enhances the selectivity of RuPOP to different tumor cells and normal cells by biotin/ receptor-mediated targeting action, realizes good anti-tumor effect .Bio-cage@Ru, accelerates the localization of RuPOP to the nucleus, and the DNA structure is cleaved under the action of DNase I to promote the release of RuPOP. In the nude mouse model of tumor-bearing mice, the Bio-cage@Ru increased the accumulation of RuPOP in the tumor area, thus improving the anti-tumor effect in vivo. In addition, the Bio-cage@Ru obviously reduces the toxicity of heart, liver, lung and kidney caused by RuPOP, and achieves the goal of high efficiency and low toxicity. In the fifth chapter, the thesis summarizes the work of the whole thesis, and puts forward the future research direction. The work of this thesis provides a solution to the problems of radiation therapy, poor chemotherapy effect, obvious toxic and side effect and drug resistance in cancer treatment.
【学位授予单位】：暨南大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R91;R730
，
本文编号：2252696
[Abstract]:With the development of nano science and technology, the application of nano drug delivery system provides an effective way for diagnosis and treatment of tumor. The nano drug loading system can integrate the drug and the imaging agent into the nano particles, and is targeted to the tumor tissue through the enhanced osmotic effect or the active targeting effect, thereby improving the treatment effect and reducing the toxic and side effects on the normal tissues. In many nano-drug delivery systems, since the biological macromolecule nano-drug carrier system has good biocompatibility and biodegradability, toxicity caused by long-term accumulation in vivo can be avoided. Based on the advantages of biodegradable nano materials, this paper aims at improving the diagnosis and treatment effect of tumor, and develops a biological macromolecule nano medicine loading system based on bovine serum albumin, phycoerythrin and DNA, which is applied to targeted therapy and image tracing of tumors. torsion resistance and radiotherapy/ chemotherapy co-treatment, etc. The main research work is as follows: The first chapter introduces the development status of nano-drugs in tumor diagnosis and treatment, and summarizes the categories, advantages and applications of biological macromolecule nano-carriers in tumor diagnosis and treatment. In the second chapter, in order to improve the effect of radiotherapy, folic acid-targeted albumin nano-drug delivery system was synthesized and the organic selenium derivative PSeD was loaded, and the albumin nano-drug delivery system (FA-BSANP@Se) with tumor targeting and biocompatibility was constructed. The combined X-ray can greatly improve the active oxygen level in the uterine cervix cancer cell and cause large-scale DNA damage, so that the G2/ M cycle block and the apoptosis signal are cascade activated, and finally the formation of the cell clone community of the cervical cancer cell is inhibited. In order to overcome the possible immune source of heterogenic BSANPs to human body, a new type of nano-carrier of phycoerythrin nanoparticles (PCNPs) as adriamycin (DOX) was prepared in the third chapter, and targeted molecular folic acid (Folate) was loaded. FA) to explore its role in reversing adriamycin resistance in tumor cells and its related mechanisms. Pharmacokinetic studies indicate that PCNPs significantly prolong the blood half-life of DOX, release DOX.FA-PCNP@DOX in acidic lysosomal environments to recognize tumor-resistant cells RHep G2 through FA/ FARs-mediated targeting, and cause mitochondrial breakage by up-regulation of intracellular reactive oxygen levels, and then blocking the activity of the ATP-dependent ABC transporter, remarkably improving the absorption and staying efficiency of DOX in the RHepG2 cells, and finally inducing the apoptosis of the RHepG2 cells. In vivo imaging experiments prove that the FA-PCNP@DOX can target tumor regions and reduce the toxicity of liver, lung and kidney caused by DOX while effectively inhibiting the proliferation of tumor bodies. Further, in order to improve the blood compatibility and targeted anti-tumor effect of metal complexes, we constructed the vector of cGVHD-targeting phycoerythrin nanoparticles as the carrier of polychrome complex (RuPOP) in the fourth chapter, and found that the cRGD-PCNP@Ru can improve the hydrophilicity of RuPOP. and prevents the interaction of RuPOP and red blood cells, thereby improving the blood compatibility of RuPOP. By carrying out active screening on a series of cervical cancer cells Caski, Siha, and Caski cells, it is found that the cRGD-PCNP@Ru can recognize the high-expression cervical cancer cell Caski through the targeting action mediated by the caspase-whole, and the accumulation of the RuPOP in the Caski tumor ball is improved, thereby improving the inhibiting effect of RuPOP on the proliferation of Caski tumor balls. In order to improve the loading efficiency and anti-tumor activity of the nano-drug delivery system and the metal complex, Biotin-targeted DNA images nano-drug delivery system was constructed as the carrier (Bio-Cage@Ru) of the metal complex RuPOP in chapter V to study its potential in improving the antitumor activity of RuPOP and reducing toxicity. Bio-Cage, as a tetrahedral cage type structure with tumor targeting property, enhances the selectivity of RuPOP to different tumor cells and normal cells by biotin/ receptor-mediated targeting action, realizes good anti-tumor effect .Bio-cage@Ru, accelerates the localization of RuPOP to the nucleus, and the DNA structure is cleaved under the action of DNase I to promote the release of RuPOP. In the nude mouse model of tumor-bearing mice, the Bio-cage@Ru increased the accumulation of RuPOP in the tumor area, thus improving the anti-tumor effect in vivo. In addition, the Bio-cage@Ru obviously reduces the toxicity of heart, liver, lung and kidney caused by RuPOP, and achieves the goal of high efficiency and low toxicity. In the fifth chapter, the thesis summarizes the work of the whole thesis, and puts forward the future research direction. The work of this thesis provides a solution to the problems of radiation therapy, poor chemotherapy effect, obvious toxic and side effect and drug resistance in cancer treatment.
【学位授予单位】：暨南大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R91;R730
，
本文编号：2252696
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