基于NMR指纹谱相关分析的生物类似药高级结构评价

建立了基于1H NMR指纹技术的生物类似药高级结构相似性分析方法，利用相关系数r和决定系数R2定量评估生物类似药与其原研药的结构相似度。将该方法应用在一种分子量为1620的抗菌性脂肽达托霉素上，比较了其类似药的活性原料成分(API)、类似药制剂与原研药克必信的高级结构相似性。实验结果显示，3个达托霉素样品的谱图存在显著差异，表明达托霉素类似药与其原研药在水溶液中存在结构差异。将该方法应用在分子量为145423的单克隆抗体药物曲妥珠单抗的高级结构比对上。对1个批次的赫赛汀原研药、4个批次的类似药的1H NMR谱进行相似性分析，相关系数r和决定系数 R2的结果显示，曲妥珠单抗的类似药与赫赛汀原研药相似度很高；进一步采用局部相关系数进行谱图分析，得到了曲妥珠单抗类似药与赫赛汀原研药之间的微小差异。研究表明，有效结合NMR指纹分析技术和统计分析方法，将在生物类似药质量控制中发挥重大的作用。

Higher Order Structure Assessment of Biosimilars Based on the Correlation of NMR Spectral Fingerprints

A procedure based on the1H NMR fingerprints of biologics was established for the assessment of higher order structure of biosimilar samples. The global correlation coefficientsr andR2 of binned NMR spectra were used to quantify structural similarity between biosimilars and originators, and a local correlation analysis method was used to find out the specific regions that contribute to the structure differences. This method was applied to the quantitative assessment of daptomycin, a lipopeptide antibiotic with a molecular weight of 1 620, by comparing the higher order structure of the biosimilar active pharmaceutical ingredient (API), its injection and originator injection Cubicin. The results showed that there were significant spectral differences among daptomycin samples, indicating structural differences between daptomycin originator and its biosimilars. Local correlation analysis further revealed the specific spectral regions that contributed to these differences. The method was also applied to a monoclonal antibody drug Trastuzumab, which is a large protein with a molecular weight of 145 423.1H NMR spectra of the originator drug Herceptin and four lots of its biosimilar were recorded and binned for statistical analysis. The correlation coefficientsr andR2 showed that Trastuzumab biosimilars seem to be highly similar to Herceptin. These results demonstrated that the combination of NMR fingerprints and statistical analysis is a powerful and robust technique in biosimilar quality control.