吉非替尼壳聚糖鱼精蛋白肺靶向纳米粒的制备及靶向性初步研究

发布时间：2018-03-11 11:10

本文选题：吉非替尼　切入点：星点设计-效应面法　出处：《锦州医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的吉非替尼是一种新型治疗非小细胞肺癌药物,为选择性表皮生长因子受体酪氨酸激酶(该酶常见于上皮来源的实体瘤)抑制剂,从而阻断肿瘤之间的信息传递,能抑制肿瘤细胞的生长、转移和血管生成,促进肿瘤细胞凋亡。目前国内市售的吉非替尼制剂为片剂,口服吸收慢,生物利用度低且伴有消化系统方面的不良反应,在血浆中容易被代谢为低活性物质失去临床作用。纳米粒作为一种纳米级微观药物载体传输系统,具有生物膜的透过性,透过肿瘤血管在肿瘤组织聚集,即高通透性和滞留效应,并能保护药物免于在血浆中代谢失活提高了其生物利用度。本实验以无毒、无刺激、具有良好的生物相容性及生物可降解性的壳聚糖和鱼精蛋白为载体原料,制备吉非替尼壳聚糖鱼精蛋白纳米粒,以改善吉非替尼在血浆中的分布代谢,降低给药剂量,提高生物利用度,增加靶组织内聚集性,减少不良反应从而获得更好的疗效。方法结合壳聚糖为载体材料制备纳米粒相关文献及预实验结果,本实验采用离子胶凝法,以壳聚糖和鱼精蛋白为载体材料,制备吉非替尼壳聚糖鱼精蛋白纳米粒。建立体外分析方法并进行方法学考察。以包封率和载药量为评价指标,通过星点设计-效应面法优化处方。对最优处方制备的载药纳米粒进行质量评价,包括外观、形态、粒径分布、表面电位、体外释放度及稳定性的考察。依据相关文献建立小鼠血浆及组织中吉非替尼高效液相色谱检测方法,并对色谱条件进行方法学考察。以吉非替尼溶液作为对照品,吉非替尼壳聚糖鱼精蛋白纳米粒为实验制剂,以小鼠为受试动物,进行组织分布实验,测定各组织及血浆中吉非替尼浓度并计算相应的药代动力学参数,进而对肺靶向吉非替尼壳聚糖鱼精蛋白纳米粒进行靶向性初步研究。结果本课题建立的体外分析方法符合方法学的要求。采用星点设计-效应面法优化的吉非替尼壳聚糖鱼精蛋白纳米粒的最优处方为:吉非替尼的质量浓度为1mg·m L-1,壳聚糖的质量浓度为3.5 mg·m L-1,鱼精蛋白的质量浓度为1 mg·m L-1。最优处方验证结果表明所建立的数学模型具有良好的预测效果。质量评价表明该纳米制剂为结构完整、大小均一稳定的具有蓝色乳光的胶体溶液,为球形或类球形实体粒,平均粒径为169 nm,Zeta电位为+23.7 m V,在p H=5.8处具有明显的缓释效果且较高的累积释药率,在4℃环境下能稳定地存储。本课题成功建立了小鼠体内高效液相检测方法,体内分布实验表明,与吉非替尼口服液相比,纳米粒在小鼠的肝、脾、肺等组织浓度较高,肺组织浓度最高。结论制备吉非替尼壳聚糖鱼精蛋白纳米粒的方法简单成熟,处方优化后制备的纳米粒是结构完整、大小均一稳定的球形或类球形实体微球,粒径分布集中,表面荷正电,体外溶出度具有明显的缓释效果,稳定性好。鼠体内组织分布实验证明,与吉非替尼口服液相比,纳米粒注射液除了具有明显的缓释作用外,还具有明显的肺组织靶向性,从而能够改善吉非替尼在体内的组织分布。
[Abstract]:Objective to gefitinib is a new medicine for treating non-small cell lung cancer, selective epidermal growth factor receptor tyrosine kinase (the enzyme found in solid tumors of epithelial origin) inhibitors, which block the transmission of information between the tumor, can inhibit tumor cell growth, metastasis and angiogenesis, promote the apoptosis of tumor cells at home. The commercially available gefitinib formulations for tablets, oral absorption is slow, bioavailability is low and the adverse reactions associated with the digestive system, the plasma can easily be metabolized to active substances. The clinical effect of low loss of nanoparticles as a nano micro drug carrier transmission system, with biological membrane permeability, through tumor blood vessels in tumor tissue is aggregation, high permeability and retention effect, and can protect the drug from metabolic inactivation in plasma to improve its bioavailability. In this experiment, non-toxic, no stimulation, good Good biocompatibility and biodegradable chitosan and protamine as carrier materials, preparation of gefitinib chitosan protamine nanoparticles to improve gefitinib in the plasma distribution and metabolism, decrease the dosage, improve the bioavailability, increase the cohesion of the target tissue in effect, reduce adverse reactions in order to obtain better methods. Combined with chitosan nanoparticles preparation of related literatures and preliminary experimental results as the carrier material, this experiment adopts ion gelation method, using chitosan and protamine as carrier materials, preparation of gefitinib chitosan protamine rice. Establish and Bai satisfied method was studied in vitro by analytical method. The encapsulation efficiency and drug loading as evaluation index, through the ccd-rsm optimization prescription. To evaluate the quality of the optimal prescription preparation of drug loaded nanoparticles including appearance, morphology, particle size distribution, surface potential, in vitro release Study on dissolution and stability. On the basis of the relevant literature established in plasma and tissues of mice by gefitinib HPLC method, and the chromatographic conditions were investigated. The gefitinib solution as the reference, gefitinib chitosan protamine nanoparticles as experimental preparation in mice by the test animal, tissue distribution experiment, determination of the plasma and tissues of gefitinib concentration and pharmacokinetic parameters were calculated accordingly, then the lung targeting gefitinib chitosan protamine nanoparticles were targeted. Preliminary research results of this paper establish in vitro analysis method in line with the methodological requirements. The ccd-rsm optimization of gefitinib optimal formulation of imatinib chitosan protamine nanoparticles: gefitinib concentration, 1mg m L-1, the concentration of chitosan was 3.5 Mg - M L-1, the quality of protamine Concentration of 1 mg m L-1. to verify the results of the optimal prescription has a good forecast results show that the established mathematical model. The quality evaluation showed that the nano preparation for structural integrity, uniform size and stable colloidal solution with blue opalescence, as spherical or spherical solid particles, the average particle size is 169 nm, the Zeta potential of +23.7 m V, in P H=5.8 has obvious sustained-release effect and higher cumulative drug release rate, can be stably stored in 4 C environment. We successfully established a HPLC method for detection of mice, showed that the distribution in vivo experiments, and gefitinib oral liquid compared to nanoparticles in mouse liver. The spleen, lung tissue was higher, the highest concentration of lung tissue. Conclusion the preparation method of gefitinib imatinib chitosan protamine nanoparticles is simple and mature, optimized preparation of nanoparticles is spherical or spherical structure is complete, uniform and stable physical size The microspheres, particle size distribution, surface is positively charged, with obvious sustained-release effect, in vitro dissolution stability. Tissue distribution experiments show that compared with Kyrgyzstan non imatinib oral liquid, injection in addition to nanoparticles had obviously sustained effect, also has the obvious to the target lung tissue, which can improve Kyrgyzstan gefitinib in vivo tissue distribution.

【学位授予单位】：锦州医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R943

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