Research based on messenger RNA (mRNA) has been a gamechanger for healthcare, boosted by the launch of the first COVID-19 vaccine in December 2020—just a year after the first reported case of the disease. mRNA-based products are now moving rapidly beyond infectious disease prophylaxis, and support from emerging companies like ExploRNA Therapeutics will be vital to the growth of the area.
Cutting-edge capping reagents
ExploRNA Therapeutics’ co-founders Jacek Jemielity (CEO) and Joanna Kowalska (CTO) created the company based on their ground-breaking research at the University of Warsaw. Supported by researchers with years of experience in chemistry, biology, and immunology, the company is developing technology that aims to increase and optimize protein expression for new and better therapeutics and vaccines. ExploRNA has strong intellectual property (IP), four products on the market, and a number of projects in its pipeline. Its approach is validated by it being the first eastern European company to receive a grant from the Bill & Melinda Gates Foundation.
At the core of ExploRNA is its library of over 150 cap analogs, based on decades of experience in mRNA synthesis. These analogs are chemically modified derivatives of the naturally occurring cap structures at the 5′ end of the mRNA in all eukaryotic cells.
In vivo, the cap structures play a role in RNA stabilization, pre-mRNA splicing, initiation of mRNA translation, and the cellular transport of mRNAs and small nuclear RNAs (snRNAs). ExploRNA’s library of diverse di-, tri-, and tetra-nucleotide derivatives of these structures, which are compatible with various in vitro transcription (IVT) conditions and templates, can be used to develop more efficient and cost-effective reagents, IVT protocols, and IVT-related molecular tools, and enzymes for research. They can also be used to create and optimize mRNA structures, and build an additional layer of IP protection for therapies and technologies. Each cap analog in the library has data associated with it, and the materials can be ordered online via the company’s website.
Working towards a shared vision
Collaborations are a powerful way for companies to access expertise and resources. ExploRNA partners with academic institutions and biopharma companies to develop mRNA therapeutics and prophylactics. Projects so far have included vaccines, protein-replacement therapies, gene-editing-based therapeutics, and cell therapies. Contract development and manufacturing organizations also value ExploRNA for the support in solving problems for their clients.
An example of an ongoing relationship is ExploRNA’s work with Wacker, a German provider of specialist products for the pharma, biotechnology, food, and nutraceutical industries.
“ExploRNA’s technologies allow us to offer our customers an effective alternative to other technologies, increasing the options for their research. They are a good collaborative partner to work with—they support us, provide the test materials we need, are available for troubleshooting, and keep us updated on new developments,” said Hagen Richter, head of nucleic acids research at Wacker. “We gain from each other.”
ExploRNA provides access to its technologies through a number of different collaboration models, ranging from purchasing capping reagents under free research licenses to exclusive therapy co-development agreements.
“Our partners may want to increase protein yield or reduce the amount of cap needed, and we work together to design and optimize the new cap analogs and test them in vitro and in vivo. The results can also help us to improve our products to support future research,” said Jemielity.
Creating unique mRNA technology
ExploRNA’s leading products are the proprietary AvantCap AG, ExploCap, AvantCap Q1, and AvantCap Q2. The AvantCap analogs enhance mRNA activity in vitro and increase the expression of therapeutic proteins in vivo compared with natural cap1, which is a currently dominating solution in the market.
AvantCap AG, a modified version of the cap1 structure, delivers high capping efficiency and produces low levels of double-stranded RNA (dsRNA) during in vitro transcription. The modification also helps with the separation of capped and uncapped mRNA fractions and impurities via high-performance liquid chromatography (HPLC). In a mouse model, AvantCap AG increased human erythropoietin (hEPO) production by between two- and six-fold with modified and unmodified uridines, respectively (Fig. 1). This improved protein expression has potential to reduce the dose of mRNA therapeutics required for patients.
Fig. 1 | Next-generation cap analogs boost protein expression. AvantCap increased human erythropoietin (hEPO) production up to two-fold when using modified N1-MeΨ messenger RNA (mRNA) and up to six-fold when using natural uridine-5′-triphosphate (UTP) compared with Cap1 in an in vivo study of C57BL/6 mice. Serum was tested for hEPO levels with an enzyme-linked immunosorbent assay (ELISA). U, uridine.
The second-generation AvantCap tetra-nucleotide cap analogs, AvantCap Q1 and Q2, are robust and have high capping efficiency and improved IVT parameters at a lower cap concentration. AvantCap Q2 is the first modified cap analog on the market to implement the cap2 structure.
ExploCap delivers high IVT yields and capping efficiencies, equivalent to unmodified cap1 analogs, and is a cost-effective alternative to the industry standard.
“Our goal is to build relationships and create better mRNA products. We believe that our technology platform has the potential to harness the power of mRNA not only in antiviral vaccines but also in more advanced applications,” said Jemielity.