Organic Solvent Nanofiltration (OSN): A New Technology Platform for Liquid-Phase Oligonucleotide Synthesis (LPOS)
Organic Solvent Nanofiltration (OSN): A New Technology Platform for Liquid-Phase Oligonucleotide Synthesis (LPOS)
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DOI：
10.1021/acs.oprd.6b00139
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发表时间：
2016-08-01
影响因子：
3.4
通讯作者：
Livingston, Andrew G.
中科院分区：
文献类型：
--
作者：
Kim, Jeong F.;Gaffney, Piers R. J.;Livingston, Andrew G.
有机溶剂纳滤（OSN）技术是一种在恶劣的有机介质中进行分子分离的膜工艺。然而，尽管 OSN 技术的潜在应用已有充分记录，但开发障碍阻碍了 OSN 技术的广泛采用。最有前途的应用领域之一是作为迭代合成平台，例如寡核苷酸或肽，其中每个合成周期后都需要彻底的纯化步骤，最好是在相同的工作溶剂中。在这项工作中，我们报告了一项使用 OSN 技术进行液相寡核苷酸合成 (LPOS) 的工艺开发研究。基于寡核苷酸（oligo）的药物正在被开发为在蛋白质表达水平发挥作用的新一代疗法。目前，超过 100 种基于寡核苷酸的药物正在进行临床试验，这表明很快就有必要以每年公吨的规模生产寡核苷酸。然而，目前还没有能够批量生产 >10 kg 寡核苷酸的合成平台。通过这里开发的工艺，我们成功地进行了八次链延伸迭代循环，并合成了 5 聚体和 9 聚体 2'-O-甲基寡核糖核苷酸硫代磷酸酯，所有这些都在液相介质中进行。本文讨论了该流程开发过程中面临的预期和意外的关键挑战，并提出了缩短开发周期的解决方案。已经进行了经济分析，强调了 LPOS-OSN 工艺的潜在竞争力和溶剂回收装置的必要性。
Organic solvent nanofiltration (OSN) technology is a membrane process for molecular separation in harsh organic media. However, despite having well-documented potential applications, development hurdles have hindered the widespread uptake of OSN technology. One of the most promising areas of application is as an iterative synthesis platform, for instance for oligonucleotides or peptides, where a thorough purification step is required after each synthesis cycle, preferably in the same working solvent. In this work, we report a process development study for liquid-phase oligonucleotide synthesis (LPOS) using OSN technology. Oligonucleotide (oligo) based drugs are being advanced as a new generation of therapeutics functioning at the protein expression level. Currently, over 100 oligo based drugs are undergoing clinical trials, suggesting that it will soon be necessary to produce oligos at a scale of metric tons per year. However, there are as yet no synthesis platforms that can manufacture oligos at >10 kg batch scale. With the process developed here, we have successfully carried out eight iterative cycles of chain extension and synthesized 5-mer and 9-mer 2'-O-methyl oligoribonucleotide phosphorothioates, all in liquid phase media. This paper discusses the key challenges, both anticipated and Unexpected, faced during development of this process and suggests solutions to reduce the development period. An economic analysis has been carried out, highlighting the potential competitiveness of the LPOS-OSN process and the necessity for a solvent recovery unit.