This project will develop a flexible and scalable RNA-manufacturing process and delivery system for the sustainable manufacture of biologic drugs, with considerably reduced material-use/energy-costs and the need for repurposing or rebuilding facilities. RNA is the medicine platform of the future with many application-types in clinical trial. RNA therapeutics are suitable for manufacture without using cell-based processes, so it is possible to take an RNA sequence to a candidate vaccine or therapeutic in weeks. A particular advantage is that same manufacturing plant can be used to produce an extraordinary array of drugs from personalised medicines to vaccines. The aim of this application is to further enhance the RNA-manufacturing process to reduce waste and improve sustainability. We will achieve this through several RNA-manufacturing innovations. We will optimise a scalable RNA Biofoundry (that we call Biofoundry in a box --BiaB). This will enable globally distributed-manufacturing via continuous-flow processes, to improve RNA purity and reduce waste. We will also develop tunable and thermostable formulations; these reduce cold-chain requirements and the need for energy-consuming ultralow temperature freezers. Alongside the equipment, novel excipients will be developed for a supramolecular _delivery system_ _that does not require organic solvents_ for sustainable manufacture of local, disease-strain-specific therapeutic/vaccine applications. We will also develop a low-loss fill-finish line so that all of the process intensification advantages of the RNA Biofoundary to produce a minimum viable product size for early clinical development are not wasted through the traditional approaches to demonstrating quality and sterility assurance. Whilst we anticipate that RNA can be used for a wide range of indications, the immediate use case is as a vaccine. Humanity has never been at greater risk of zoonotic-pathogen outbreaks, recent high-profile examples include zoonotic viral pathogens (SARS/MERS/Ebola/Influenza/monkeypox). The COVID-19 pandemic demonstrated the capacity for rapid development, high efficacy and positive safety profile of mRNA vaccines as needs arise. The focus of this application therefore will be to optimise the sustainability of facility capable of manufacturing a pipeline of RNA vaccines to prevent a potential avian-influenza crossover-event before it becomes a pandemic. Improved pathogen surveillance and sequencing will enable infectious diseases to be identified and therapeutics designed earlier (e.g. the CEPI 100-day mission). The scale of the manufacture we are proposing, combined with the thermostability makes our approach highly suitable for use in low-and-middle-income countries (LMICs), where there is a vaccination backlog.

RNA-based medicines and vaccines are of increasing importance to UK national health and wealth. These therapeutics are effective at low dose levels. Different diseases can be treated with different RNAs; but Different RNAs, however, may be made by the same process. The RNA performs its therapeutic task inside a patient's cells: intracellular delivery is currently achieved by incorporating RNA into Lipid Nanoparticles (LNPs). However, LNP use is challenging because of high lipid cost and patent constraints: solvent use (then removal) in LNP manufacture; and expensive cold chain between LNP manufacture and patient. We have developed a new delivery system that uses safe and accessible small-molecule components which, with CB\[8\] and RNA, spontaneously self-assemble. These supramolecular RNA therapeutics (SMRTs) efficiently encapsulate and deliver RNA into cells, do not require organic solvents, have low material costs (<£2/dose), good room-temperature stability, and avoid the toxicity of polymer-based RNA-delivery systems. SMRTs are adaptable to different RNA types and may be formulated to include special stabilising or cell-targeting components, giving potential to address diseases in different parts of the body. Our innovation will combine SMRT formulations with development of a miniaturised flow-based manufacture unit to a scale suitable for in-hospital applications. The facility (currently aimed at producing millions of doses of RNA-vaccine/day) uses modular continuous-flow processes, which permits scale independence (down to individual RNA-therapeutic doses). The processes have already demonstrated in-situ in a single unit manufacture of high-purity mRNA at high concentration _and_ formulation into a delivery system. Integrating multi-RNA synthesis with SMRT formulation enables streamlined production of multiple drug products from the same facility. Ease in scaling enables longer running with minimal adjustments, while continuous process-verification-in-flow reduces validation complexity and simplifies regulatory reporting, yielding shorter approval schedules and time-to-market. Small-footprint units can be sited in individual hospitals, enabling local production of RNA-medicine for individual patients or stratified patient groups, eliminating cold- and supply-chain hurdles. The delivery-system/manufacturing-unit combination will be developed for the clinical example of triple negative breast cancer (TNBC). In the UK, 51% of cancer diagnoses are for breast cancer (BC), which has 7% mortality. Compared to other BCs, TNBC tends to grow and spread faster, have fewer treatment options, more recurrences, and worse outcomes.so needs new therapeutic options. BC is an NHS (and global) priority. Project partners UK SMEs Aqdot and Centillion Technology and Nottingham University's research groups in Pharmacy and Medicine combine expertise in supramolecular-assembly, flow-based manufacture, and cancer therapy delivery/design/testing.