Li Yangjie, Hu Yanyang, Logsdon David L, Liu Yong, Zhao Yuejie, Cooks R Graham
560 Oval Drive, Department of Chemistry, Purdue University, West Lafayette, IN, 47907-1393, USA.
Dept. of Analytical Sciences, MRL, Merck & Co., Inc., 126 East Lincoln Ave, Rahway, NJ, 07065, USA.
Pharm Res. 2020 Jul 10;37(7):138. doi: 10.1007/s11095-020-02868-y.
Forced degradation is critical to probe the stabilities and chemical reactivities of therapeutic peptides. Typically performed in bulk followed by LC-UV or LC-MS analysis, this traditional workflow consists of a reaction/analysis sequence and usually requires half a day to several days to form and measure the desired amounts of degradants. A faster method is needed to study peptide degradation in a shorter time in order to speed up the drug development process.
In the new rapid method developed in this study, peptide degradation occurs in levitated aqueous microdroplets using the Leidenfrost effect.
This two-minute reaction/analysis workflow allows major degradation pathways of Buserelin, Octreotide, Desmopressin and Leuprorelin to be studied. The reactions include deamidation, disulfide bond cleavage, ether cleavage, peptide bond hydrolysis, and oxidation.
The accelerated forced degradation method requires a minimal amount of therapeutic peptide per stress condition, and the appropriate extent of degradation can be readily generated in seconds by adjusting the droplet levitation time. Levitated microdroplets should be applicable in pharmaceutical development to rapidly determine the intrinsic stability of therapeutic peptides and to aid formulation development by screening the effects of excipients on the stability of the peptides. Graphical abstract.
强制降解对于探究治疗性肽的稳定性和化学反应性至关重要。这种传统的工作流程通常是批量进行，随后进行液相色谱 - 紫外（LC - UV）或液相色谱 - 质谱（LC - MS）分析，它由一个反应/分析序列组成，通常需要半天到几天的时间来形成和测量所需量的降解产物。为了加快药物研发进程，需要一种更快的方法在更短时间内研究肽的降解。
在本研究开发的新的快速方法中，利用莱顿弗罗斯特效应使肽在悬浮的水微滴中发生降解。
这个两分钟的反应/分析工作流程能够研究布舍瑞林、奥曲肽、去氨加压素和亮丙瑞林的主要降解途径。这些反应包括脱酰胺、二硫键断裂、醚键断裂、肽键水解和氧化。
加速强制降解方法在每种应激条件下所需的治疗性肽量极少，通过调整微滴悬浮时间，可在数秒内轻松产生适当程度的降解。悬浮微滴应可应用于药物研发，以快速确定治疗性肽的固有稳定性，并通过筛选辅料对肽稳定性的影响来辅助制剂研发。图形摘要。
