LC-MS法在治疗性寡核苷酸药物检测中的应用

Application of LC-MS in the analysis of therapeutic oligonucleotide drugs

治疗性寡核苷酸(oligonucleotides,OGNs)药物是人工化学合成的短单链或双链核酸,长度通常为15~30个碱基对。OGNs作为一种新的治疗药物正在迅速发展,并在各种疾病领域的药物发现和开发中受到越来越多的关注。与欧美相比,除Spinraza(nusinersen)作为孤儿药在中国获批上市外,暂无其他OGNs药物在国内上市。国内OGNs药物开发起步较晚,仍然处于发展初期,但由于国内患者群体基数较大,需求较多,未来伴随OGNs药物开发的持续推进,以及国内企业相应技术的逐步成熟,我国OGNs药物市场有望迎来快速发展。OGNs药物由于其独特的理化性质,生物分析方法开发难度大,目前,国内关于OGNs药物定量分析方法的报道还很少。开发一种灵敏可靠的方法来表征生物样品中的OGNs是研究其药代动力学和药效学性质的关键。相对于传统的ELISA法,LC-MS法可以同时定量完整OGNs及其代谢物,特别是高分辨质谱的广泛应用不仅可以提供目标OGNs的定量信息,还可以对代谢物进行鉴定,提供碱基组成、序列结构等信息,目前已成为OGNs定量分析的首选方法。本文主要描述了LC-MS在治疗性OGNs药物检测中的应用,并阐述了其优点和局限性,最后探讨了LC-MS用于检测OGNs的发展趋势,即更低的检测水平和潜在的通用方法。

Therapeutic oligonucleotides(OGNs)drugs are artificially synthesized single or double stranded short nucleic acids,typically 15 to 30 base pairs in length.OGNs have been rapidly developed as new therapeutic drugs with increasing attention in the discovery and development of drugs concerning various disease fields.Compared with Europe and America,there are currently no other OGNs drugs listed in China,except for Spinraza,which has been approved for marketing as an orphan drug.The development of OGNs in China started relatively late and is still in its early stages of development.However,the OGNs drug market in China is anticipated to grow quickly due to the country's large population,high patient demand,ongoing support for the development of oligonucleotide drugs in the future,and the steady maturation of related technologies by domestic businesses.Because of their special physicochemical characteristics,OGNs drugs are challenging to design biological analysis techniques.Currently,there are few reports on quantitative analysis methods for oligonucleotide drugs in China.Therefore,the development of sensitive and reliable bioanalysis methods for oligonucleotides is the key to investigate oligonucleotides'pharmacokinetic and pharmacodynamic properties.Liquid chromatography-mass spectrometry(LC-MS)can quantify OGNs and their metabolites concurrently,compared with traditional ELISA approaches.Numerous benefits come with using LC-MS,in particular,the extensive use of high-resolution mass spectrometry allows for the identification of metabolites,which provides details on base composition and sequence structure,in addition to quantitative information about target oligonucleotides.It has now emerged as the go-to technique for OGN quantitative analysis.The application of LC-MS in the identification of therapeutic oligonucleotide medicines is the primary focus of this paper,which also discusses its benefits and drawbacks.Lastly,it looks at the LC-MS development trend for oligonucleotide detection,which includes a lower detection level and potential general methods.