Automated Production of RALA/Nucleic Acid Nanoparticles from Bench to Patient Dose

RNAi therapeutics have the potential to transform healthcare interventions as evidenced by the approval of 2 products in the last 2 years for life threatening diseases. RNAi therapy is designed to transiently reduce a defective gene for therapeutic purposes. It is a rapidly growing market with 109 RNAi based therapeutics in clinical trials (July 2018) \[Wu X. and Turnball A.P. 2018\]. However, there are still issues that surround the RNAi therapeutics which include getting to the appropriate tissue and then ensuring intracellular delivery to the destination site. Recent studies have also indicated that those with underlying health conditions such as diabetes, high-blood pressure or smokers have an increased number of ACE-2 receptors in the lung epithelium \[Leung J.M, 2020\]. Studies have revealed that COVID-19 uses the ACE-2 receptor to enter cells in order to produce more viral particles that can infect more ACE-2 receptor positive cells \[Kuba K. 2005\]. The ACE-2 receptor plays a role for many biological functions but if expression could be lowered for a short period of time it could reduce the infectivity of the virus and help tip the balance towards healthy recovery. RNAi could be used to transiently reduce expression of this ACE-2 receptor but only if there is an appropriate delivery system. pHion (Belfast SME) have developed a solution for RNAi delivery that is safe, does not further exacerbate the immune system, preferentially delivers the therapeutic to the lung and is cost-effective, ultimately enabling widespread adoption of the RNAi therapy. The innovation centres around the use of a peptide termed RALA that is designed to condense RNAi into nanoparticles (NPs) that have the properties necessary to cross cell membranes, escape endosomes delivering the cargo to the cytoplasm with high efficiency. The NPs formed between the RNAi which is designed to reduce ACE-2 expression and the RALA peptide NPs do not require cold chain storage and can be stored for many months without losing functionality. However, we do not as yet have a methodology in place to support the large-scale production of these NPs. Indeed, for the nucleic acid industry, one of the greatest hurdles will be the manufacture of novel therapeutics. Therefore with clear alignment to the specific theme of challenges as a result of COVID-19, this project is designed to accelerate and optimise the scale-up of the RALA/RNAi therapeutic to patient doses in order to be 'future ready'. The proposed 9 month project is designed to develop the optimal conditions for the automated production of functional NPs using microfluidics that can be readily transferred to clinical doses. We will also develop the optimal lyophilisation process to ensure a highly stable functional product. Finally, with regulatory framework in place and proof that we can transfer our process externally to scale up to clinical doses, we will be well positioned to take this therapy to the clinic and to position RALA as the go-to delivery system for RNAi therapeutics to the lung.