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

基于信使核糖核酸（mRNA）的治疗方法通过沉默特定基因或过度表达治疗性蛋白来发挥作用，其潜力现已被证明是辉瑞和Moderna COVID-19疫苗的基础。由于母体分子的不稳定性和不期望的物理化学性质，将寡核苷酸治疗药物递送到患者体内受到限制，因此通过将其掺入脂质纳米颗粒（LNP）来实现疗效。LNPs的直径通常在50-100纳米之间，由聚乙二醇化的脂质组成，聚乙二醇化的脂质有助于控制颗粒大小，二asteraroyl磷脂酰胆碱有助于形成纳米颗粒的结构，胆固醇也有助于形成纳米颗粒的结构。最重要的是，它们还含有一种可电离的阳离子脂质，它包裹着带负电荷的寡核苷酸，有助于增加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，设计量身定制的可电离阳离子脂质，将其纳入针对COVID和/或RNA癌症治疗的最佳LNP递送系统中。在合理的方法下，将制备一系列修饰的可电离脂质，并将其纳入LNP中并装载特定的寡核苷酸，然后进行理化性质研究，以确定粒径、包封量和pKa（包括单个脂质和LNP系统）。在特定选择的测定法中进行效力评估（将在适当的课程项目中确认）和随后的稳定性研究之后，将生成结构活性关系概况，该概况将用于推动迭代合成活动，以找到最佳LNP系统。特定的技能,学生将获得博士学位的一部分下面列出:标准和并行合成高度functionalised脂质moleculesCharacterisation和分析脂质使用核磁共振,LCMS, IR, MALDI,,8经,CHNPurification脂质分子和他们的前体,通过flash列chromatographyPreparation oligo-lipid纳米粒子系统通过微观流体/ micromixerAnalysis LNP系统的物理化学性质如Z-average, PDI,电动电位,pKa测定、DLS和低聚物包封/吸附效率。

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.