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一个准确而快速的P qNMR测定方法用于寡核酸治疗药物

Jiayi Li¹, Fu Chen², Deyi Zhang³

¹Division of Pharmaceutical Quality Research II, Office of Pharmaceutical Quality Research, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, Maryland 20993, United States.

Analytical chemistry

|October 11, 2024

概括

一种新的定量NMR (qNMR) 方法可以准确量化寡核酸治疗药物,克服UV-260nm分析的局限性. 这种31P qNMR方法为寡核酸药物和参考标准提供了可靠的初级测定.

科学领域:

分析化学分析化学
生物化学生物化学
制药科学制药科学

背景情况:

氧核酸治疗药物需要精确的质量保证测试.
传统的UV-260nm量化是不准确的,因为结构修改和更高阶结构 (HOS).
重力测量分析受到水和 counterions 的影响,影响准确性.

研究的目的:

开发一个快速,准确和灵活的定量测定对寡核酸治疗药物.
为了比较一种新的31P定量-NMR (qNMR) 方法与UV-260nm和重力测量分析的性能.
评估31P qNMR作为对寡核类药物的主要测定方法的适用性.

主要方法:

开发一个外部引用的31P定量-NMR (qNMR) 试验,运行时间小于1小时.
使用单链DNA标准验证qNMR方法.
对完整的寡核酸药物 (eteplirsen, inotersen, inclisiran) 的31P qNMR和UV-260nm方法进行比较分析.

主要成果:

31P qNMR测定显示了DNA标准的UV-260nm的1-5%的结果.
与药物标签相比,31P qNMR的低估度为7 ± 2% (eteplirsen),8 ± 1% (inotersen) 和12 ± 1% (inclisiran).与药物标签相比,31P qNMR的低估度为7 ± 2% (eteplirsen),8 ± 1% (inotersen) 和12 ± 1% (inclisiran).
紫外线-260nm低估的度显着更高:28 ± 3% (eteplirsen),10 ± 3% (inotersen) 和10 ± 1% (inclisiran),这表明灭绝系数存在问题,特别是对于PMO结构.

结论:

31P qNMR 方法提供了精确可靠的核酸治疗药物的量化.
该方法克服了UV-260nm测试的局限性,特别是对改性寡核酸的测试.
31P qNMR被提议作为寡核酸药物和参考标准的主要测定方法,提供更高的准确性,效率和灵活性.

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