Solution‐Phase Synthesis of Short Oligo‐2′‐deoxyribonucleotides by Using Clustered Nucleosides as a Soluble Support

Kungurtsev, Vyacheslav; Laakkonen, J.; Molina, Alejandro Gimenez; Virta, Pasi

doi:10.1002/ejoc.201300864

Cited by 24 publications

(39 citation statements)

References 35 publications

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“…Pentaerythritol-based (Kungurtsev et al, 2013;Molina et al, 2015;Molina et al, 2014) 5-mer (20 nucleotides, 4-arm) P (×2) 1.5 eq/branch (DNA/RNA) (Adaman-1-yl)acetyl (de Koning et al, 2006) 6-mer (6 nucleotides, 1-arm) E (×8) 1.5 eq. (DNA)…”

Section: Of 17mentioning

confidence: 99%

“…During the last decade, various chemical protocols for LPOS have been a subject of significant interest due to the increasing demand for large quantities of therapeutic ONs. This effort includes extraction (de Koning et al, 2006), precipitation (Donga et al, 2006;Kim et al, 2013;Kungurtsev et al, 2013;Matsuno et al, 2016;Molina et al, 2015;Molina et al, 2014;Schwenger et al, 2017), chromatography (Molina et al, 2012), and OSNbased (Gaffney et al, 2015;Kim et al, 2016) purification techniques for scale up of ONs. Although some of these methods provide possible avenues for large-scale LPOS, none have been demonstrated at scales useful for the commercial production of ONs.…”

Section: Summary and Future Predictions Toward Green Manufacturing Ofmentioning

confidence: 99%

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Liquid‐Phase Oligonucleotide Synthesis: Past, Present, and Future Predictions

Molina

Sanghvi²

2019

CP Nucleic Acid Chemistry

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Therapeutic oligonucleotides have emerged as a powerful paradigm with the ability to treat a wide range of the human diseases. As a result, we have witnessed more than one hundred oligonucleotides currently in active clinical trials and eight Food and Drug Administration (FDA)‐approved drugs. Until now, the demand for oligonucleotide‐based drugs has been fulfilled by conventional solid‐phase synthesis in an effective manner. However, there are products in advanced stages of clinical trials projecting a collective demand of metric ton quantities in the near future. Therefore, large‐scale manufacturing of these products has become a high priority for process chemists. This article summarizes the advances in liquid‐phase oligonucleotide synthesis (LPOS) as a possible alternative strategy to meet the scale‐up challenge. A review of the literature describing major efforts in developing LPOS technologies is presented. Gratifyingly, serious attempts are under way to develop an efficient environmentally benign green chemistry protocol that is scalable and cost effective for the manufacturing of oligonucleotides. A summary of the most innovative LPOS protocols has been included to provide a glimpse of what may be possible in the future for large‐scale production of oligonucleotides. © 2019 by John Wiley & Sons, Inc.

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Section: Of 17mentioning

confidence: 99%

Section: Summary and Future Predictions Toward Green Manufacturing Ofmentioning

confidence: 99%

Liquid‐Phase Oligonucleotide Synthesis: Past, Present, and Future Predictions

Molina

Sanghvi²

2019

CP Nucleic Acid Chemistry

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“…Owing to the symmetrical structure of the support, NMR and mass spectroscopic characterization is possible at any stage of the chain assembly. The 3´-terminal nucleoside was immobilized to this support as a 3´- O -(4-pentynoyl) derivative by Cu(I)-catalyzed 1,3-dipolar cycloaddition [ 43 ]. This support is soluble in MeCN and dioxane but precipitates quantitatively in MeOH.…”

Section: Reviewmentioning

confidence: 99%

Synthesis of oligonucleotides on a soluble support

Lönnberg¹

2017

Beilstein J. Org. Chem.

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Oligonucleotides are usually prepared in lab scale on a solid support with the aid of a fully automated synthesizer. Scaling up of the equipment has allowed industrial synthesis up to kilogram scale. In spite of this, solution-phase synthesis has received continuous interest, on one hand as a technique that could enable synthesis of even larger amounts and, on the other hand, as a gram scale laboratory synthesis without any special equipment. The synthesis on a soluble support has been regarded as an approach that could combine the advantageous features of both the solution and solid-phase syntheses. The critical step of this approach is the separation of the support-anchored oligonucleotide chain from the monomeric building block and other small molecular reagents and byproducts after each coupling, oxidation and deprotection step. The techniques applied so far include precipitation, extraction, chromatography and nanofiltration. As regards coupling, all conventional chemistries, viz. phosphoramidite, H-phosphonate and phosphotriester strategies, have been attempted. While P(III)-based phosphoramidite and H-phosphonate chemistries are almost exclusively used on a solid support, the “outdated” P(V)-based phosphotriester chemistry still offers one major advantage for the synthesis on a soluble support; the omission of the oxidation step simplifies the coupling cycle. Several of protocols developed for the soluble-supported synthesis allow the preparation of both DNA and RNA oligomers of limited length in gram scale without any special equipment, being evidently of interest for research groups that need oligonucleotides in large amounts for research purposes. However, none of them has really tested at such a scale that the feasibility of their industrial use could be critically judged.

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“…[17,[19][20][21] Recently ad iscrete, non-polymeric synthesis support was developed in which four oligo chains were grown simultaneously around ap entaerythritol core, and the productsw ere then precipitated from methanol. [22,23] Whilst approaches to oligo synthesis based upon precipitation or crystallisation are in theory scalable, it is questionable whether this would be truly practical. Process development of industrial precipitation is often time-consuming and labour-intensive, with the conditions being unique to each compound.…”

Section: Introductionmentioning

confidence: 99%

Liquid‐Phase Synthesis of 2′‐Methyl‐RNA on a Homostar Support through Organic‐Solvent Nanofiltration

Gaffney

Kim

Valtcheva

et al. 2015

Chemistry A European J

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Due to the discovery of RNAi, oligonucleotides (oligos) have re-emerged as a major pharmaceutical target that may soon be required in ton quantities. However, it is questionable whether solid-phase oligo synthesis (SPOS) methods can provide a scalable synthesis. Liquid-phase oligo synthesis (LPOS) is intrinsically scalable and amenable to standard industrial batch synthesis techniques. However, most reported LPOS strategies rely upon at least one precipitation per chain extension cycle to separate the growing oligonucleotide from reaction debris. Precipitation can be difficult to develop and control on an industrial scale and, because many precipitations would be required to prepare a therapeutic oligonucleotide, we contend that this approach is not viable for large-scale industrial preparation. We are developing an LPOS synthetic strategy for 2′-methyl RNA phosphorothioate that is more amenable to standard batch production techniques, using organic solvent nanofiltration (OSN) as the critical scalable separation technology. We report the first LPOS-OSN preparation of a 2′-Me RNA phosphorothioate 9-mer, using commercial phosphoramidite monomers, and monitoring all reactions by HPLC, 31P NMR spectroscopy and MS.

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Solution‐Phase Synthesis of Short Oligo‐2′‐deoxyribonucleotides by Using Clustered Nucleosides as a Soluble Support

Cited by 24 publications

References 35 publications

Liquid‐Phase Oligonucleotide Synthesis: Past, Present, and Future Predictions

Liquid‐Phase Oligonucleotide Synthesis: Past, Present, and Future Predictions

Synthesis of oligonucleotides on a soluble support

Liquid‐Phase Synthesis of 2′‐Methyl‐RNA on a Homostar Support through Organic‐Solvent Nanofiltration

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