New Mexico Patent of the Month – May 2024
The quest for effective RNA-based therapeutics and diagnostics has led to groundbreaking innovations in nucleotide synthesis. Among these innovations, Nature’s Toolbox, Inc. (NTx) emerges with a revolutionary system for the production of pseudouridine compounds, a critical component in stabilizing RNA molecules.
At the heart of NTx’s invention lies a meticulously designed system comprising isolated uracil nucleobase and ribose-5-phosphate. These substrates undergo a series of enzymatic and chemical transformations orchestrated to produce pseudouridine-5′-monophosphate (ΨMP), the cornerstone of NTx’s method.
Central to the process is pseudouridine-5′-phosphate glycosidase (PsuG), an enzyme catalyst that facilitates the conversion of uracil nucleobase and ribose-5-phosphate into ΨMP. PsuG, derived from either thermophilic or enteric bacteria, catalyzes this transformation with remarkable efficiency, laying the foundation for subsequent steps.
Following PsuG’s enzymatic action, a protecting agent, often hexamethyldisilazane (HMDS), reacts with ΨMP, forming a protected ΨMP. This protective step ensures the stability of the compound throughout the synthesis process, safeguarding against undesired reactions.
Next, a methylating agent, typically iodomethane, reacts with the protected ΨMP, yielding a protected N1-methyl-pseudouridine-5′-monophosphate (protected m1ΨMP) compound. This methylation step introduces critical modifications essential for the desired properties of the final product.
Subsequently, a deprotection agent, commonly consisting of a combination of ammonia and methanol, removes the protecting groups, resulting in the formation of N1-methyl-pseudouridine-5′-monophosphate (m1ΨMP). This deprotection step unveils the active form of the compound, poised for further modification.
Finally, a phosphorylating agent, often polyphosphate kinase (PPK2) sourced from thermophilic bacteria like Deinococcus geothermalis, catalyzes the sequential phosphorylation of m1ΨMP, culminating in the production of N1-methyl-pseudouridine-5′-diphosphate (m1ΨDP) and N1-methyl-pseudouridine-5′-triphosphate (m1ΨTP). This phosphorylation process imparts crucial functionalization, rendering the final product ready for therapeutic applications.
NTx’s innovative system revolutionizes the production of pseudouridine compounds, addressing critical bottlenecks in RNA-based therapeutics and diagnostics. By combining enzymatic and chemical methodologies with precise substrate manipulation, NTx paves the way for a new era of RNA-focused innovations, promising transformative advancements in healthcare and beyond.
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