The success of mRNA-based COVID-19 vaccines has generated interest in RNA-based drug development, with over 200 RNA drugs estimated to be in development. The promise of RNA medicines in turn has engendered innovation in manufacturing these drugs, including from start-up companies.

By Patricia Van Arnum, Editorial Director, DCAT, pvanarnum@dcat.org

Enzymatic RNA synthesis
A relatively new company focused on improving RNA-based manufacturing is Watertown, Massachusetts-based EnPlusOne Biosciences, which was founded in 2022 as a spinout from the Wyss Institute for Biologically Inspired Engineering at Harvard University. The company has a proprietary enzymatic RNA oligonucleotide synthesis technology, which was used to make the antisense strand of a commercially approved small interfering RNA (siRNA) drug, Leqvio (inclisiran), a treatment for hypercholesteremia (high cholesterol) licensed from Alnylam Pharmaceuticals by Novartis.

The standard method of chemically synthesizing RNA is limited in terms of the range of nucleotides that can be incorporated in the RNA molecule and also requires the use of solvents in the manufacturing process. The company’s proprietary aqueous-based, template-independent enzymatic RNA oligonucleotide synthesis platform is used as an alternative to traditional chemical methods. The aqueous-based system allows for reduced solvent use. The company reported in August (August 2024) the synthesis of a Leqvio antisense strand, an enzymatically-synthesized 23-nucleotide (23-nt) sequence fully modified with 2’-F and 2’-OMe nucleotide modifications along with two leading and trailing phosphorothioate backbone modifications. The company says it represents the first known template-independent enzymatic synthesis of a fully natural siRNA length oligonucleotide (23-nt).

EnPlusOne is also part of a collaboration led by the Wyss Institute that was awarded up to $27 million by the Advanced Research Projects Agency for Health (ARPA-H), an agency within the US Department of Health and Human Services that supports biomedical and health research. The agreement, announced in August (August 2024) covers multi-disciplinary efforts to develop a disease-agnostic novel RNA therapeutic with the potential to treat multiple diseases, including types of cancer and infectious diseases. Initially, up to $3.5 million of the funding is focused on EnPlusOne’s enzymatic platform.

The ARPA-H award will allow the Wyss Institute team to accelerate and expand efforts to advance the therapy toward an investigational new drug submission to the US Food and Drug Administration (FDA). The team is initially focusing on cancer and will explore the platform’s potential for difficult-to-treat infectious diseases. The ARPA-H project will build on a newly developed Duplex RNA technology and use RNA delivery capabilities developed by groups at the Wyss Institute, along with the enzymatic RNA synthesis capabilities of EnPlusOne, to optimize the RNA’s design and scalability.

Codexis, a Redwood City, California-based enzyme engineering company, which provides biocatalysts for small-molecule drug manufacturing, has leveraged its expertise in enzymatic processes to RNA manufacturing through its ECO Synthesis manufacturing platform for enzymatic RNA synthesis. A second RNA platform, Codec HiCap RNA Polymerase, is an engineered, co-transcriptional capping RNA polymerase for produceing synthetic mRNA. Last December (2023), Codexis partnered with Aldevron, a CDMO of plasmid DNA, RNA, and proteins, under which Aldevron acquired a global exclusive license to Codexis’ Codex HiCap RNA Polymerase. Under the deal, Aldevron will receive global manufacturing and commercialization rights to the Codex HiCap RNA Polymerase, and Codexis will receive payments for near-term technical milestones, along with commercial milestones and sales-based royalties.

Continuous mRNA manufacturing
Like the production of many other biologics, the current production of mRNA is batched and requires many steps that can create bottlenecks in its production. Continuous mRNA manufacturing has the potential to mitigate those bottlenecks and help meet the growing demand for mRNA material and make it easier to ramp up production quickly, for example, in the event of a pandemic or other public health emergency.

FDA plays a role in facilitating the development of new manufacturing technology through the Emerging Technology Program under the Center for Drug Evaluation and Research (CDER) and through the Advanced Technologies Program under the Center for Biologics and Research (CBER).

CBER’s Advanced Technologies Program provides an interactive mechanism to promote dialogue, education, and input between CBER and prospective innovators/developers of advanced manufacturing and testing technologies. In addition, CBER has awarded several grants and contracts to support research projects to study and recommend improvements for the advanced manufacturing of biological products, including the investigation and development of innovative monitoring and control techniques. The funded research addresses knowledge and experience gaps identified for emerging manufacturing technologies and ​supports​ the development and adoption of such technologies in the biological product sector.

In this context, a consortium led by researchers at the Massachusetts Institute of Technology (MIT) announced last year (2023),  a three-year, $82-milion award from FDA, through CBER, to develop a pilot-scale system for the first fully integrated, continuous mRNA manufacturing platform. 

With the funding from FDA, MIT has formed the Center for Continuous mRNA Manufacturing, led by Richard Braatz, the Edwin R. Gilliland Professor in MIT’s Department of Chemical Engineering and the principal investigator for the project. The engineering challenges will be tackled by researchers at MIT as well as collaborators at Penn State University, led by Professor Andrew Zydney, and Rensselaer Polytechnic Institute, led by Professors Steven Cramer and Todd Przybycien. A substantial portion of the project has been subcontracted to ReciBioPharm, the biologics business unit of Recipharm, a Stockholm, Sweden headquartered CDMO. 

The new research program builds on the success of the Novartis–MIT Center for Continuous Manufacturing, a $85-million program that ran from 2007 to 2019, which developed and demonstrated the first bench-scale integrated continuous manufacturing system of both the API and drug product, a so-called “blue-sky vision” for continuous manufacturing.  

The main goal of the work of the Center for Continuous mRNA Manufacturing is to advance the field of mRNA therapeutics by providing a continuous manufacturing template for companies to follow while facilitating collaboration throughout the biopharmaceuticals industry. The research team will also work closely with FDA to ensure the pilot-scale system adheres to cGMP and informs regulatory strategies. MIT is focusing on experimental innovation, and ReciBioPharm is implementing the attenable and GMP-level manufacturing program. Work at MIT and ReciBioPharm is already underway to develop automation and advanced controls for quality assurance and improve midstream processing.

RNA delivery
Overcoming the challenge of RNA delivery has also engendered deals in the RNA space as larger bio/pharma companies seek to access specialized technologies. Last year (2023), Novartis acquired DTx Pharma, a San Diego, California-based preclinical stage biotechnology company developing siRNA therapies to build its neuroscience pipeline and xRNA platform capabilities, in a $1-billion deal ($500 million upfront and $500-million in potential milestones). The deal included DTx-1252, a potential therapy for the neuromuscular disorder, Charcot-Marie-Tooth disease type 1A (CMT1A), and two additional preclinical programs for other neuroscience indications. The acquisition also netted Novartis with DtxPharm’s Fatty Acid Ligand Conjugated OligoNucleotide (FALCON) platform, which enables the delivery and activity of siRNA therapeutics to tissues beyond the liver, enhancing biodistribution and cellular uptake.

Ethris, a Munich, Germany-based company developing mRNA therapeutics, has a proprietary, non-immunogenic messenger RNA (SNIM mRNA) and lipidoid nanoparticle (SNaP LNP) technology platforms for delivering stabilized mRNAs directly to the respiratory system via optimized formulation and nebulization technologies. In June (June 2024), Cipla, a Mumbai, India-based bio/pharmaceutical company, took an additional $3-million stake in Ethris, to add to the $15-million stake taken in 2022. Last November (November 2023), Ethris became one of the founding members of the Alliance for mRNA Medicines, a scientific and policy organization for advancing and advocating for mRNA and next-generation encoding RNA therapeutics and vaccines.