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

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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.