Modern approaches to therapeutic oligonucleotide manufacturing

Science, Journal Year: 2024, Volume and Issue: 384(6692)

Published: April 11, 2024

Therapeutic oligonucleotides are a powerful drug modality with the potential to treat many diseases. The rapidly growing number of therapies that have been approved and in advanced clinical trials will place unprecedented demands on our capacity manufacture at scale. Existing methods based solid-phase phosphoramidite chemistry limited by their scalability sustainability, new approaches urgently needed deliver multiton quantities required for therapeutic applications. community has risen challenge rethinking strategies oligonucleotide production. Advances chemical synthesis, biocatalysis, process engineering technologies leading increasingly efficient selective routes sequences. We review these developments, along remaining challenges opportunities innovations allow sustainable diverse products.

Language: Английский

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Controlled enzymatic synthesis of oligonucleotides

Communications Chemistry, Journal Year: 2024, Volume and Issue: 7(1)

Published: June 18, 2024

Abstract Oligonucleotides are advancing as essential materials for the development of new therapeutics, artificial genes, or in storage information applications. Hitherto, our capacity to write (i.e., synthesize) oligonucleotides is not efficient that read sequencing) DNA/RNA. Alternative, biocatalytic methods de novo synthesis natural modified dire need circumvent limitations traditional synthetic approaches. This Perspective article summarizes recent progress made controlled enzymatic synthesis, where temporary blocked nucleotides incorporated into immobilized primers by polymerases. While robust protocols have been established DNA, RNA XNA more challenging. Nevertheless, using a suitable combination protected and polymerase has shown promises produce even though production long DNA/RNA/XNA sequences (>1000 nt) remains We surmise merging ligase- polymerase-based would help current shortcomings synthesis.

Language: Английский

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Template-independent enzymatic synthesis of RNA oligonucleotides

Nature Biotechnology, Journal Year: 2024, Volume and Issue: unknown

Published: July 12, 2024

Abstract RNA oligonucleotides have emerged as a powerful therapeutic modality to treat disease, yet current manufacturing methods may not be able deliver on anticipated future demand. Here, we report the development and optimization of an aqueous-based, template-independent enzymatic oligonucleotide synthesis platform alternative traditional chemical methods. The is made possible by controlled incorporation reversible terminator nucleotides with common 3′- O -allyl ether blocking group using new CID1 poly(U) polymerase mutant variants. We achieved average coupling efficiency 95% demonstrated ten full cycles liquid phase produce natural therapeutically relevant modified sequences. then qualitatively assessed solid phase, performing several N + 5 controlled-pore glass support. Adoption aqueous-based process will offer key advantages including reduction solvent use sustainable manufacturing.

Language: Английский

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Synthesis of short DNA and RNA fragments by resonant acoustic mixing (RAM)

RSC Mechanochemistry, Journal Year: 2024, Volume and Issue: 1(3), P. 244 - 249

Published: Jan. 1, 2024

We demonstrate the first use of Resonant Acoustic Mixing (RAM) without bulk solvent for synthesis short oligonucleotide fragments.

Language: Английский

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Artificial nucleotide codons for enzymatic DNA synthesis

Chemical Communications, Journal Year: 2023, Volume and Issue: 59(98), P. 14547 - 14550

Published: Jan. 1, 2023

Herein, we report the high-yielding solid-phase synthesis of unmodified and chemically modified trinucleotide triphosphates (dN3TPs). These synthetic codons can be used for enzymatic DNA provided their scaffold is stabilized with phosphorothioate units. Enzymatic three rather than one letter nucleotides will useful to produce xenonucleic acids (XNAs) in vitro selection functional nucleic acids.

Language: Английский

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Oligonucleotides: evolution and innovation

Medicinal Chemistry Research, Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 21, 2024

Abstract Oligonucleotides, comprising single or double strands of RNA DNA, are vital chemical compounds used in various laboratory and clinical applications. They represent a significant class therapeutics with rapidly expanding range uses. Between 1998 2023, 19 oligonucleotides have received approval from the U.S. Food Drug Administration (FDA). Their synthesis methods undergone evolution over time. This review examines several oligonucleotide techniques, including phosphodiester, phosphotriester, phosphoramidite approaches. It begins discussion an early method involving phosphoryl chloride intermediate, which proved unstable prone to hydrolysis. The then transitions solid-phase method, uses polymer resins as solid support, emphasizing its advantages both phosphotriester techniques. is followed by exploration recent advancements enzymatic synthesis, concluding on modifications bases, sugars, backbones designed improve their properties therapeutic potential.

Language: Английский

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Template-dependent DNA ligation for the synthesis of modified oligonucleotides

Published: Dec. 6, 2023

Chemical modification of DNA is a common strategy to improve the properties oligonucleotides, particularly in context therapeutics and nanotechnology. Existing synthetic methods essentially rely on phosphoramidite chemistry or polymerization nucleoside triphosphates are limited terms size, scalability, sustainability. Herein, we report robust alternative method for de novo synthesis modified oligonucleotides using template-dependent ligation shortmer fragments. Our approach based fast scaled accessibility chemically monophosphates as substrates T3 ligase. This has shown high tolerance chemical modifications, flexibility overall efficiency, thereby granting access an ultimately broad range different lengths (20 →160 nucleotides). We have applied this clinically relevant antisense drugs highly ultramers. Furthermore, designed chemoenzymatic great potential numerous applications oligonucleotide therapeutics, bioorganic chemistry, pharmacology, biology.

Language: Английский

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Template-Independent Enzymatic RNA Synthesis

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 9, 2024

Abstract A route to prepare ribonucleoside triphosphates featuring a 3’-aminoxy (3’-O-NH 2 ) removable blocking group is reported here. We then show that versions of two DNA polymerases, human polymerase theta (Polθ) and mimiviral PrimPol, accept these as substrates add single nucleotides an RNA primer under engineered conditions. Cleaving the O-N bond in 3’-O-NH within extended regenerates 3’-OH group, facilitating subsequent cycles second, selected, nucleotide. These enzymes together enable template-independent enzymatic synthesis (TIERS) exploiting cyclic reversible termination framework. The study shows this process ready for instrument adaptation by using it three ribonucleotides Polθ. This work creates new way synthesize with de novo defined sequence, without requiring protecting groups, hazardous solvents, sensitive reagents bedevil phosphoramidite-based synthesis.

Language: Английский

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Nucleic Acids Production

Topics in medicinal chemistry, Journal Year: 2024, Volume and Issue: unknown

Published: Jan. 1, 2024

Language: Английский

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