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: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=709507
• 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 年里，我将专注于开发新的主动装载技术，将高电荷分子封装到脂质体中，以改善其递送。我将重点关注三种类型的高电荷药物：带正电的小分子、带负电的小分子和带负电的大分子。我将从每一类中选择一种模型药物，并为每一类开发一种新的主​​动装载技术。假设：离子对剂将中和化合物的电荷，以增加其膜渗透性并驱动脂质体的负载。 目标 1：开发一种主动装载方法，将带正电的小分子封装到脂质体中。我们的模型药物是庆大霉素 (GEN)，它有 5 个带正电荷的氨基。我们将开发和优化一种新的负载方法来制备脂​​质体GEN，并比较其在游离GEN和被动负载制备的脂质体GEN的小鼠体内的药代动力学（PK）。 目标 2：开发一种主动装载方法，将带负电的小分子封装到脂质体中。我们的模型药物是氯膦酸盐 (CLO)，它具有 2 个带负电荷的磷酸基团。脂质体 CLO 用于消耗动物组织巨噬细胞以生成研究模型。我们将开发和优化一种新的装载技术来制备脂质体CLO，并将其与游离CLO和被动装载制备的脂质体CLO在小鼠中的巨噬细胞消耗活性进行比较。 目标 3：开发一种主动装载方法，将带负电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核酸 (siRNA)。我们将开发一种离子对活性负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荷瘤小鼠中的生物分布（BD），并与游离siRNA和阳离子脂质体-siRNA复合物进行比较。 短期目标：开发主动装载技术，将 GEN、CLO 和 siRNA 封装到脂质体中，以改善其递送。长期目标：开发一种平台技术，将各种类型的高电荷药物加载到脂质体中，以改善其 PK 和 BD，从而提高生物利用度，并有可能创造有效的体内研究工具或药品。该项目将为 HQP 提供全面的培训，包括药物配方设计、制造和表征、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领域的学术界和工业界备受追捧，而在我的项目中接受培训的 HQP 在这些技术性工作中具有很强的竞争力。我已经发表了超过 25 篇关于脂质体技术的论文和专利，并向行业授权了 2 项技术，并有 2 种产品处于临床试验阶段。该项目将产生具有高商业化价值的新脂质体工程技术。