GlaxoSmithKline, Medicines Research Centre, Gunnels Wood Road, Stevenage, Hertfordshire, UK.
Rapid Commun Mass Spectrom. 2011 Feb 28;25(4):511-25. doi: 10.1002/rcm.4886.
Oligonucleotide-based drugs are beginning to establish themselves within the pharmaceutical industry as important agents in the treatment of various disease states with the potential of exhibiting high specificity with gene targeted therapies. Recent studies regarding RNA interference has stimulated interest in this field. There are now an increasing number of oligonucleotide-based pharmaceutical products in various stages of clinical development for the treatment of life-threatening diseases. As a result, the production of synthetic oligonucleotides has become increasingly important, with both antisense and RNAi-related oligonucleotides under development as therapeutic agents. One potential drug candidate currently under development at GlaxoSmithKline, is a 2'-O-methyl phosphorothioate in which the non-bridging oxygens of the phosphate diester are replaced with sulphur. Oligonucleotides are polymeric sequences made from an array of nucleotides (RNA, DNA and their respective analogs) usually ranging from 20-100 nucleotides. The polar nature, low thermal stability, complexity and large molecular weights of oligonucleotides have posed a challenge for the analysis of oligonucleotides by mass spectrometry. This paper demonstrates the use of negative ion electrospray with a combination of high resolution and high mass accuracy for the characterisation of oligonucleotides with the intention of supporting an evidence of structure document for a regulatory submission. This is a new area within the mass spectrometry field and as such there is limited software amongst the instrument companies for the data processing for the analysis of these compounds. Therefore, many of the examples in the literature only use mass spectrometry to generate average molecular weights by deconvoluting the multiple charged states observed to give an average molecular weight; under-utilizing the capability of high-resolution instruments.
寡核苷酸类药物作为治疗各种疾病状态的重要药物，在制药行业中开始崭露头角，具有基因靶向治疗的高特异性潜力。最近关于 RNA 干扰的研究激发了人们对这一领域的兴趣。现在，有越来越多的基于寡核苷酸的药物产品处于不同的临床开发阶段，用于治疗危及生命的疾病。因此，合成寡核苷酸的生产变得越来越重要，反义寡核苷酸和 RNAi 相关寡核苷酸都在作为治疗剂进行开发。目前，葛兰素史克公司正在开发的一种潜在药物候选物是 2'-O-甲基硫代磷酸酯，其中磷酸二酯的非桥氧被硫取代。寡核苷酸是由核苷酸（RNA、DNA 及其各自的类似物）组成的聚合物序列，通常长度为 20-100 个核苷酸。寡核苷酸的极性、低热稳定性、复杂性和大分子量给通过质谱分析寡核苷酸带来了挑战。本文展示了负离子电喷雾与高分辨率和高精度质量分析的结合，用于寡核苷酸的特征描述，旨在为监管申报的结构文件提供支持证据。这是质谱领域的一个新领域，因此仪器公司的数据分析处理软件在该领域非常有限，无法满足分析这些化合物的需求。因此，文献中的许多例子仅使用质谱通过解卷积观察到的多电荷态来生成平均分子量，从而利用高分辨率仪器的能力不足。
