Introduction DOI
Alessandra Tolomelli, Lucia Ferrazzano, Walter Cabri

et al.

Royal Society of Chemistry eBooks, Journal Year: 2024, Volume and Issue: unknown, P. 1 - 8

Published: Oct. 18, 2024

The increasing number of new therapeutic oligopeptides and oligonucleotides, commonly referred to as TIDES, approved each year has led consistent rapid growth the market size need produce large volumes these active pharmaceutical ingredients. In addition, in TIDES chemistry, experimental design be tailored for specific molecule most suitable technology often selected from among a effective approaches. development sustainable processes are therefore necessary guarantee not only optimal purity yield, but also management solvents, reagents, energy waste. This introductory chapter outlines purpose this book, namely serve valuable reference synthesis sustainability, which topics highlighted that discussed following chapters.

Language: Английский

Controlled enzymatic synthesis of oligonucleotides DOI Creative Commons
Maëva Pichon, Marcel Hollenstein

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

Citations

13
Template-independent enzymatic synthesis of RNA oligonucleotides DOI Creative Commons
Daniel J. Wiegand, Jonathan Rittichier,

Ella Meyer

et al.

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

Citations

10
Toward sustainable solid-phase peptide synthesis strategy – in situ Fmoc removal DOI Creative Commons

Ndumiso Mthethwa,

K. P. Nandhini,

Ashish Kumar

et al.

Green Chemistry Letters and Reviews, Journal Year: 2024, Volume and Issue: 17(1)

Published: March 11, 2024

Solid-phase peptide synthesis (SPPS) is the method of choice for peptides research and production purposes. Despite having several positive features, it remains a challenge to reduce amount solvent waste generated during synthesis. We proposed 3-step protocol (in-situ Fmoc removal) where washing after coupling was eliminated. Here, in-situ removal optimized by adding an extra 4-methylpiperidine (4-MP) treatment ensure Fmoc. Additionally, second addition carbodiimide step performed form in situ more active ester Fmoc-amino acid. The number washings has been kept minimum three two them 1% OxymaPure total 4-MP. This strategy, which saves up 60% solvent, successfully demonstrated important Active Pharmaceutical Ingredients (APIs), angiotensin II afamelanotide synthesis, with high purity. strategy will be added green toolbox SPPS making approach sustainable.

Language: Английский

Citations

6
Speeding up sustainable solution-phase peptide synthesis using T3P® as a green coupling reagent: methods and challenges DOI Creative Commons
Alexia Mattellone, Dario Corbisiero, Lucia Ferrazzano

et al.

Green Chemistry, Journal Year: 2023, Volume and Issue: 25(7), P. 2563 - 2571

Published: Jan. 1, 2023

A fast and greensolution-phase peptide synthesis (GSolPPS) via continuous protocol, addressed with propylphosphonic anhydride T3P® as coupling reagent N -benzyloxycarbonyl-protecting group easily removed by hydrogenation is herein reported.

Language: Английский

Citations

13
Mechanism Unraveling of Scalable Antifreeze Oligopeptides for Enhanced Cryopreservation DOI

Hongfeng Zhou,

Mengyao Song,

Xiaohu Zhang

et al.

Langmuir, Journal Year: 2025, Volume and Issue: unknown

Published: March 31, 2025

Cryopreservation is fundamental to cell-based therapeutics but severely limited by ice formation and growth, which causes irreversible membrane rupture, osmotic imbalance, chilling injury. Antifreeze proteins conventional cryoprotectants like gold standard dimethyl sulfoxide (DMSO) struggle offer a cost-effective biocompatible strategy fitting various cellular storage. Herein, we design fabricate class of oligopeptides exhibiting potent recrystallization inhibition (IRI) activity, achieving 55.5% reduction in the mean largest grain size at concentration less than 0.1 wt %. The side-chain functional groups (e.g., hydroxyl amine groups) length (<10 amino acids) are meticulously optimized avoid thermal hysteresis (TH) activity that sudden burst fatal needle-like crystals. Simulation experimental results illustrate mechanism involves binding crystal surface disrupting ordering water molecules, thereby preventing well-structured Notably, employment as maintains cell proliferation differentiation capabilities while having high viability 90–95%, comparable 10% DMSO.

Language: Английский

Citations

0
Eco-friendly synthesis, structural insights, and antimicrobial potential of 1,3,5-tri-N-substituted hexahydro-1,3,5-triazine derivatives DOI
Jyoti Kumawat, Sonika Jain,

Namita Misra

et al.

Analytical Chemistry Letters, Journal Year: 2025, Volume and Issue: unknown, P. 1 - 23

Published: March 16, 2025

Language: Английский

Citations

0
Design and application of hydroxyl ionic liquids for the supported synthesis of oligopeptides DOI
Hao Ling, Shuaijun Ding, Jianmin Chen

et al.

Tetrahedron Letters, Journal Year: 2025, Volume and Issue: unknown, P. 155645 - 155645

Published: May 1, 2025

Language: Английский

Citations

0
A Platform for Controlled Template-Independent Enzymatic Synthesis of RNA Oligonucleotides and Therapeutics DOI Open Access
Daniel J. Wiegand, Jonathan Rittichier,

Ella Meyer

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: June 29, 2023

Abstract Therapeutic RNA oligonucleotides have shown tremendous potential to manage and treat disease, yet current manufacturing methods cannot deliver on this promise. Here, we report the development optimization of a novel, aqueous-based, template-independent enzymatic oligonucleotide synthesis platform as an alternative traditional chemical methodologies. Our is made possible by reversible terminator nucleoside triphosphates enzyme capable their incorporation. We show that many common therapeutic modifications are compatible with our process demonstrate natural modified in both liquid solid phases. offers unique advantages over synthesis, including realization more sustainable produce oligonucleotides. One-Sentence Summary An novel nucleotide building blocks used synthesize template independently under aqueous conditions.

Language: Английский

Citations

9
Assessment of Sulfur Impurities in GMP-Grade Diisopropylcarbodiimide and Their Impact on Coupling Reagent-Induced Side Reactions in Peptide Synthesis DOI
Jan Pawlas,

Jon H. Rasmussen

Organic Process Research & Development, Journal Year: 2023, Volume and Issue: 27(5), P. 982 - 992

Published: May 4, 2023

Diisopropylcarbodiimide (DIC) constitutes one of the most widely used coupling reagents for amide bond formation in peptide synthesis. We have previously reported that DIC contains a varying amount sulfur (61–2030 ppm), and main S-containing impurity is thiourea DITU, which exhibits an excellent ability to suppress side reactions caused by N-oxyl radicals formed from common N-hydroxyl-coupling additives such as HOBt Oxyma (Green Chem. 2019, 21, 5990). Aiming understand how quality impacts synthesis, here, we report evaluation 14 batches GMP-grade seven different vendors. With elemental analysis (S), gas chromatography–mass spectrometry, high-performance liquid chromatography, chromatography-mass nuclear magnetic resonance, determined currently use not but rather diazetidine-2-thione [1,3-diisopropyl-4-(isopropylimino)-1,3-diazetidine-2-thione, DIDT], 2 + cycloaddition between iPr-NCS. The identity DIDT was confirmed synthesis iPr-NCS, followed spiking batch with synthesized material. In comparative DITU aimed at determining their breakdown Fmoc-Cys(Trt)-OH Oxyma, completely prevented Cys decomposition, whereas had no effect on suppression amino acid. Finally, catalytic amounts well thiols DTT NAC positive minimization HOBt-induced Fmoc-Trp(Boc)-OH, urea DIU, every DIC-mediated reaction, accelerated Trp substrate.

Language: Английский

Citations

7
Elevating 1-tert-Butyl-3-ethylcarbodiimide (TBEC) as a Reagent for Sustainable Peptide Synthesis: Quality Assessment and Minimizing Racemization, Precipitation, and Radical-Induced Side Reactions by TBEC/Oxyma Couplings in an Environmentally Sensible Solvent DOI
Jan Pawlas,

Ji-Hyung Choi,

Christoph von Bargen

et al.

Organic Process Research & Development, Journal Year: 2023, Volume and Issue: 27(7), P. 1348 - 1364

Published: June 23, 2023

The finding that the widely used peptide coupling reagents DIC and Oxyma form toxic H-CN (McFarland, A. D. Org. Process Res. Dev. 2019, 23, 2099) has prompted studies aimed at minimization, attained, for example, by solvent engineering (Erny, M. 2020, 24, 1341) substituting with TBEC (Manne, S. R. 2022, 26, 2894). Here, an integrated study of TBEC/Oxyma as couplers is reported, focusing not only on performance in couplings but also its cost, hazards associated use, sustainability route synthesis, end life strategies, well potential impact impurities reagent synthesis. TBEC/Oxyma-mediated NBP/EtOAc (1:4) proceeded minimal racemization, free precipitation, radical side reactions irrespective quality. These results hold great promise broad adoption suitable green media a strategy sustainable synthesis from R&D lab to manufacturing plant.

Language: Английский

Citations

7