纳米脂质体在肿瘤亚细胞靶器官分布动力学计算模型的设计

项目名称： 纳米脂质体在肿瘤亚细胞靶器官分布动力学计算模型的设计

项目编号： No.81202486

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 药物学、药理学

项目作者： 李颖寰

作者单位： 首都医科大学

项目金额： 24万元

中文摘要： 拟研究粒径为100-200 nm的纳米脂质体在细胞/亚细胞水平的转运动力学，设计计算机模型计算纳米脂质体在亚细胞靶器官的量变曲线，建立一种制剂筛选的新方法。细胞水平的实验研究主要分为脂质体与细胞表面的结合（B）及内吞（I）过程，设计了时间动力学和稳态浓度研究两组实验，将B按照生理学路径拆分为不饱和的非特异性表面结合（NSB）和可饱和的表面结合（SB）两种方式，建立相应的数学模型拟合实验值，得到细胞水平的转运速率常数；进一步将I拆分为不同亚细胞隔室的脂质体量，选择不同荧光探针标记亚细胞隔室（早/晚期内涵体、溶酶体、细胞核等），精密追踪脂质体在不同隔室间的转运时间及量变曲线，建模拟合得到亚细胞水平的转运速率常数。为完善模型的普适性，选择化学药物阿霉素和基因药物siRNA为两种模型药物，分别以被动靶向脂质体和主动靶向脂质体为研究对象，选择2种肿瘤细胞为实验对象，拟合实验值确定模型参数。

中文关键词： 脂质体；细胞内转运；内涵体逃逸；数学模型；

英文摘要： The goal of this study is to develop intracellular kinetic models of liposomes with size at 100-200 nm, both at cell level and at sub-cell level, which show a new method for formulation selection. Cell membrane binding (B) is separated into nonsaturable binding (NSB) and saturable binding (SB). Internalization (I) is further described as liposomes in different sub-cellular compartments, with cytosol and lysosome as the most important organelles studied. Doxorubicin and siRNA are selected as model drugs for chemical medicines and gene therapy. Passive-targeting liposomes and active-targeting liposomes are prepared respectively. Two tumor cells (e.g., MCF-7 and HL-60) are tested. Cell-associated liposomal concentration time courses with equal extracellular liposomes concentrations; and curves with different extracellular concentration at equilibrium are measured. Sub-cellular organelles, i.e., early/late endosomes, lysosomes and nuclei, are distinguished with their respective markers or pH-sensitive fluorescent probes. We will visualize the intracellular trafficking of drug-loaded liposomes in live cells by fluorescence microscopy. The observed data will be fitted in the established mathematic models of computer to get different rate constants among cellular and subcellular compartments. Finally, the established c

英文关键词： liposomes；intracellular trafficking；endosomal escape；mathematic models；