Development of active loading technologies for encapsulating highly charged molecules into liposomes

开发将高电荷分子封装到脂质体中的主动装载技术

• 批准号: RGPIN-2017-03787
• 负责人: Li, ShyhDar
• 金额: $ 2.04万
• 依托单位: University of British Columbia
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=682245
• 关键词: Development; active; loading; technologies; encapsulating

Over the next 5 years I will focus on developing new active loading technologies to encapsulate highly charged molecules into liposomes to improve their delivery. I will focus on three types of highly charged drugs: positively charged small molecules, negatively charged small molecules and negatively charged macromolecules. I will select one model drug from each class and develop a new active loading technology for each. Hypothesis: An ion-pairing agent will neutralize the charge of a compound to increase its membrane permeability and drive the loading into liposomes.******Obj 1: Develop an active loading method to encapsulate a positively charged small molecule into liposomes. Our model drug is gentamicin (GEN) that has 5 positively charged amino groups. We will develop and optimize a new loading method to prepare liposomal GEN, and will compare its pharmacokinetics (PK) in mice with free GEN and liposomal GEN prepared with passive loading.******Obj 2: Develop an active loading method to encapsulate a negatively charged small molecule into liposomes. Our model drug is clodronate (CLO) that has 2 negatively charged phosphate groups. Liposomal CLO is used to deplete tissue macrophages in animals to generate research models. We will develop and optimize a new loading technology to prepare liposomal CLO, and will compare its macrophage depleting activity in mice with free CLO and liposomal CLO prepared with passive loading. ******Obj 3: Develop an active loading method to encapsulate a negatively charged macromolecule into liposomes. Our model drug is small interfering ribonucleic acid (siRNA) used to study gene function. We will develop an ion-pairing active loading method to fabricate liposomal siRNA. We will assess the interaction of the liposomal siRNA with blood components and cells and its biodistribution (BD) in tumor-bearing mice, and compare with free siRNA and cationic liposome-siRNA complex.******Short term goal: Develop active loading technologies to encapsulate GEN, CLO and siRNA into liposomes to improve their delivery. Long term goal: Develop a platform technology to load various types of highly charged drugs into liposomes to improve their PK and BD for increased bioavailability, with potential to create effective in vivo research tools or pharmaceutical products. This program will offer comprehensive training for HQP, including pharmaceutical formulation design, fabrication and characterization, in vitro cell based and molecular assays and in vivo PK/BD studies. These knowledge and skills are highly sought after in academia and industry in the pharmaceutical area, and HQP trained in my program have been highly competitive for these skilled jobs. I have published >25 papers and patents with liposomal technologies and licensed 2 technologies to industry with 2 products in clinical trials. This program will yield new liposomal engineering technologies with high commercialization values.**

在接下来的5年中，我将专注于开发新的主动加载技术，以将高电荷分子封装到脂质体中以改善其递送。我将专注于三种高电荷的药物：带积极的小分子，带负电荷的小分子和带负电荷的大分子。我将从每个班级中选择一种模型药物，并为每个班级开发一种新的主​​动加载技术。假设：一个离子对剂将中和化合物的电荷，以增加其膜渗透性并将载荷驱动到脂质体中。我们的模型药物是庆大霉素（Gen），具有5个带电的氨基群。我们将开发并优化一种制备脂质体生长的新负载方法，并将其在小鼠中的药代动力学（PK）与用被动载荷制备制备的自由生长的药代动力学（PK）。我们的模型药物是氯膦酸盐（CLO），该药物具有2个带负电的磷酸基团。脂质体CLO用于耗尽动物的组织巨噬细胞以产生研究模型。我们将开发并优化一种新的加载技术来制备脂质体CLO，并将其在小鼠中的巨噬细胞耗尽活性与免费的CLO和用被动载荷制备的脂质体CLO进行比较。 ****** OBJ 3：开发一种主动加载方法，将带负电荷的大分子封装到脂质体中。我们的模型药物是用于研究基因功能的小干扰核糖酸（siRNA）。我们将开发一种离子配对的主动加载方法来制造脂质体siRNA。我们将评估脂质体siRNA与血液成分和细胞的相互作用及其在肿瘤小鼠中的生物分布（BD），并与免费的siRNA和阳离子脂质体 - siRNA复合物进行比较。********短期目标。长期目标：开发一种平台技术，将各种高度充电的药物加载到脂质体中，以改善其PK和BD以提高生物利用度，并有可能创建有效的体内研究工具或药品。该计划将为HQP提供全面的培训，包括药物配方设计，制造和表征，基于体外细胞和分子测定以及体内PK/BD研究。这些知识和技能在制药区的学术界和工业中受到了极大的追捧，在我的计划中接受培训的HQP对这些熟练的工作具有很高的竞争力。我已经通过脂质体技术发表了> 25篇论文和专利，并在临床试验中使用2种产品获得了2种工业的技术。该计划将产生具有高商业化值的新脂质体工程技术。**