Design and development of new Lipid Nano Particle delivery systems for new RNA-based c therapeutics: A rationally designed chemistry and microfluidics

设计和开发新型脂质纳米粒子递送系统，用于新型基于 RNA 的 c 疗法：合理设计的化学和微流体

• 批准号: 2889386
• 金额: --
• 依托单位: University of Strathclyde
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2889386
• 关键词: Design; development; new; Lipid; Nano

The potential of messenger ribonucleic acid (mRNA) based therapeutics which exhibit their effect by the silencing of specific genes or overexpression of therapeutic proteins has now been proven as basis of the Pfizer and Moderna COVID-19 vaccines. As delivery of oligonucleotides therapeutics into the patient is limited due to the instability and undesirable physicochemical properties of the parent molecules, efficacy is achieved by incorporation into a lipid based nanoparticle (LNP). LNPs typically, have diameters of between 50-100 nm and are comprised of a PEGylated lipid that helps control particles size, diasteraroylphosphatidylcholine to help form the structure of the nanoparticle and cholesterol which also contributes to the structure. Most importantly, they also contain an ionisable cationic lipid that encapsulates the negatively charged oligonucleotide and helps to increase the stability of the RNA. The approach in this project is to combine synthetic lipid chemistry with microfluidics to design tailor made ionisable cationic lipids to be incorporated into optimum LNP delivery systems for COVID and/or RNA cancer therapeutics. In a rational approach, a series of modified ionisable lipids will be prepared, incorporated into the LNP and loaded with the specific oligonucleotide before undergoing physicochemical property investigations to determine particle size, encapsulation loading and pKa (both individual lipid and LNP system). Following potency evaluation in specific chosen assays (to be confirmed during the course project as appropriate) and subsequent stability studies, a structure activity relationship profile will be generated which will be used to drive forward an iterative synthesis campaign to find the optimum LNP systems.The specific technical skills that the student will gain as part of this PhD are listed below:Standard and parallel synthesis of highly functionalised lipid moleculesCharacterisation and analysis of lipids using NMR, LCMS, IR, MALDI, HRMS, CHNPurification of lipid molecules and their pre-cursors, by flash column chromatographyPreparation of oligo-lipid nano particle systems via microfluids/micromixerAnalysis of physicochemical properties of LNP systems eg Z-average, PDI, zeta-potential, pKa determination, DLS and oligo encapsulation/adsorption efficiency.

基于信使核糖核酸 (mRNA) 的疗法的潜力已被证明是辉瑞 (Pfizer) 和 Moderna COVID-19 疫苗的基础，这种疗法通过特定基因的沉默或治疗性蛋白质的过度表达来发挥作用。由于母体分子的不稳定性和不良理化性质，寡核苷酸治疗剂向患者的递送受到限制，因此通过掺入基于脂质的纳米颗粒（LNP）来实现疗效。 LNP 的直径通常在 50-100 nm 之间，由有助于控制颗粒大小的聚乙二醇化脂质、有助于形成纳米颗粒结构的二硬脂酰磷脂酰胆碱和有助于形成纳米颗粒结构的胆固醇组成。最重要的是，它们还含有可电离的阳离子脂质，可封装带负电荷的寡核苷酸，有助于提高 RNA 的稳定性。该项目的方法是将合成脂质化学与微流体相结合，设计定制的可电离阳离子脂质，将其纳入用于 COVID 和/或 RNA 癌症治疗的最佳 LNP 输送系统中。在合理的方法中，将制备一系列修饰的可电离脂质，将其掺入 LNP 中并加载特定的寡核苷酸，然后进行物理化学性质研究以确定颗粒大小、封装负载和 pKa（单独的脂质和 LNP 系统）。在特定选择的测定中进行效力评估（在适当的课程项目期间确认）和随后的稳定性研究后，将生成结构活性关系图谱，该图谱将用于推动迭代合成活动以找到最佳的 LNP 系统。作为该博士学位的一部分，学生将获得的具体技术技能如下：高功能化脂质分子的标准和平行合成使用 NMR、LCMS、IR、MALDI、HRMS、CHN 表征和分析脂质脂质分子的纯化及其前体，通过快速柱色谱通过微流体/微混合器制备低聚脂质纳米粒子系统分析LNP系统的理化性质，例如Z平均、PDI、zeta电位、pKa测定、DLS和低聚封装/吸附效率。