Biocatalytic Approaches to Therapeutic Oligonucleotide Manufacturing
Therapeutic oligonucleotides bind to mRNA to modulate the production of disease related proteins and have emerged as a potential new drug modality for the treatment of a whole range of disease areas. However, current methods of chemical synthesis are not scalable and current marketed therapies are generally limited to the treatment of rare diseases. To synthesise high volume oligonucleotide products required for the treatment of more common diseases, more scalable and sustainable methods are required. Our research focusses on developing biocatalytic approaches to oligonucleotide synthesis using engineered DNA modifying enzymes. Our research is support by the UKRI, Nucleic Acid Therapeutic Accelerator, Medicines Manufacturing Innovation Centre, and multiple industrial partners.
Featured publications:
Moody, E. R.; Obexer, R.; Nickl, F.; Spiess, R.; Lovelock, S. L. “An Enzyme Cascade Enables Production of Therapeutic Oligonucleotides in a Single Operation”. Science 2023, 380 (6650), 1150–1154. https://doi.org/10.1126/science.add5892.
Obexer, R.; Nassir, M.; Moody, E. R.; Baran, P. S.; Lovelock, S. L. “Modern Approaches to Therapeutic Oligonucleotide Manufacturing”. Science 2024, 384 (6692), eadl4015. https://doi.org/10.1126/science.adl4015
Wild-type enzymes are often not suitable for direct use in pharmaceutical manufacturing and must first undergo optimization to improve properties such as substrate specificity, selectivity and stability. Directed evolution is a powerful and versatile technology for adapting enzymes to improve their efficiency under process conditions. The MIB is a world-leading interdisciplinary research centre with state-of-the-art infrastructure for accelerated enzyme evolution, including automated liquid handling robots, a colony picker and a high throughput analytical facility. We employ a range of medium to ultra-high throughput screening techniques in order to engineer biocatalysts for sustainable and cost-effective manufacturing of pharmaceuticals.
Featured publication:
A. J. Burke, W. R. Birmingham, Y. Zhuo, B. Zucoloto da Costa, R. Crawshaw, T. W. Thorpe, I. Rowles, J. Finnigan, C. Young, S. J. Charnock, S. L. Lovelock*, N. J. Turner*, A. P. Green* “An Engineered Cytidine Deaminase for Biocatalytic Production of a Key Intermediate of the Covid-19 Antiviral Molnupiravir” J. Am. Chem. Soc. 2022 144, 3761-3765.
Engineering Enzymes for Sustainable Pharmaceutical Manufacturing
The combination of computational design and directed evolution is a powerful approach to create enzymes for non-biological reactions. However, the range of chemistries accessible is restricted by Nature’s alphabet of canonical amino acids, which contain limited functionality and are not optimal for many desirable biocatalytic transformations. Genetic code expansion allows the site-selective installation of ‘chemically-programmed’ non-canonical amino acids (ncAAs) into enzyme active sites, which provide opportunities to access organocatalytic mechanisms not represented in Nature. Our research focusses on engineering translation components to allow incorporation of new functional residues into proteins. We then design and engineer proteins containing these ncAAs, to deliver catalysts for a broad range of chemical transformations.
Featured publications:
R. Crawshaw, A. E. Crossley, L. Johannissen, A. J. Burke, S. Hay, C. Levy, D. Baker, S. L. Lovelock*, A. P. Green*, “Engineering an Efficient and Enantioselective Enzyme for the Morita-Baylis-Hillman Reaction” Nature Chem. 2022, 14, 313-320.
A. Burke†, S. L. Lovelock†, A. Frese, R. Crawshaw, M. Ortmayer, M. Dunstan, C. Levy, A. P. Green, ‘’Design and evolution of an enzyme with a non-canonical organocatalytic mechanism’’ Nature 2019, 570, 219.
