Manufacturing Challenge: Engineering enzymes to scale up nucleoside analogue manufacturing

A team of researchers from the University of Manchester—part of the NATA-funded BioManNAT (Manufacturing Challenge) research consortium—has achieved a significant breakthrough in the production of nucleoside analogues, critical components of many nucleic acid therapeutics. Their innovative approach, described in a recent paper published in Nature Synthesis, uses engineered enzymes to carry out synthesis in a faster, cleaner, and more sustainable way. 

Nucleoside analogues are modified versions of the natural building blocks of DNA and RNA, playing a critical role in treating viral infections and genetic diseases, and enhancing the stability and performance of therapeutic oligonucleotides. However, their manufacturing remains a major bottleneck, often requiring complex, multi-step chemical syntheses that involve protecting group strategies, harsh reagents, and lengthy purification processes. This makes production time-consuming, costly, and difficult to scale.

The Manchester team tackled this challenge by developing a biocatalytic method for synthesising 2′-functionalised nucleoside analogues. They engineered variants of the enzyme DERA (deoxyribose-5-phosphate aldolase) to expand its ability to accept a broader range of chemical building blocks. These improved enzymes were then integrated into a one-pot biocatalytic cascade, allowing the multi-step synthesis to proceed in a single vessel, without the need for intermediate purification or protecting groups. This work draws on the expertise of Professor Nick Turner’s group, which led the enzyme engineering and synthesis of the modified DERA variants, and Dr Sarah Lovelock’s team, which developed the one-pot reaction system and contributed to process optimisation.

Using this platform, the researchers successfully synthesised a range of 2′-modified nucleoside analogues—including 2′-OH, 2′-Me, and 2′-F forms of adenosine and 2’-OH guanosine—with high selectivity and efficiency. The process was scaled up to preparative levels, suitable for further development, highlighting its potential for industrial application.

This enzymatic approach has direct relevance to the supply of raw materials for nucleic acid therapeutics, offering a scalable and flexible route to intermediates such as phosphoramidites and nucleoside triphosphates. These modified nucleosides are critical for the manufacture of oligonucleotide therapies, which often require gram-scale quantities for development and multi-kilo scale quantities for production. By avoiding time-consuming protection strategies while preserving high stereoselectivity, the biocatalytic method provides efficient access to these essential building blocks. Its streamlined, scalable design not only helps ease supply chain pressures, but also reduces environmental impact and lowers the barrier to large-scale, sustainable manufacturing of complex oligonucleotides. 

“What’s exciting about this approach is not just the efficiency, but its potential to make the production of oligonucleotide building blocks more sustainable. It offers a new route that could ease supply pressures while reducing environmental impact.”, said Barrie Cassey, Technical Director, Medicines Manufacturing Innovation Centre, CPI

The work represents a significant step towards the sustainable and flexible manufacturing of nucleic acid therapeutics. For organisations like NATA, this breakthrough offers a promising pathway to accelerate the development and scalable production of advanced nucleic acid technologies, ultimately benefiting patients worldwide.

Read the full article: An engineered aldolase enables the biocatalytic synthesis of 2′-functionalized nucleoside analogues

Learn more about the Manufacturing Challenge: NATA Manufacturing Challenge: BioManNAT

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