Assessing the Behaviour of Endotoxin in Biopharmaceutical Formulations: Lowering the Likelihood of Drug-Induced Toxic Shock in Patients

生物制药是治疗各种疾病的增长最快的治疗类型，具有巨大的个性化药物潜力。注射给患者的生物药物制剂应不含污染内毒素(脂多糖)，否则可能会引起不必要的热原反应，更严重的情况可能会导致感染性休克和死亡。脂多糖是一种大的两亲性分子，由脂肪酸成分(脂类A)和多糖成分组成，含有磷酸盐和氨基葡萄糖。这就产生了亲水的“外组分”、离子核和亲水的“内”组分；这意味着它可以在超过临界浓度(临界胶束浓度，CMC)的情况下形成胶束。LLP可能会与制剂成分相互作用，如离子、缓冲液和生物制药本身，这可能会使它们的检测和随后的去除复杂化。内毒素单体和胶束与制剂的不同成分之间的特异性和非特异性相互作用的性质尚未得到研究。这项拟议的项目旨在开发新的方法来监测LLP分子在溶液中的行为，并表征它们与二价阳离子、螯合剂、缓冲液和单抗的相互作用，并探索这些分子簇的形成方式。该项目的发现最终将有助于提高将新的、更安全的治疗方法推向市场的速度，并降低救命药物的成本。该项目将在高级溶液核磁共振技术、LLPS化学和其他生物物理方法(如CG-MALS、DLS和A4F)方面提供广泛的培训，以全面了解该系统中的分子间和分子内相互作用。每年至少有一个月的时间将在阿斯利康位于剑桥的工厂进行安置，以进行额外的实验，并获得进一步的工业工作经验。

Biopharmaceuticals are the fastest-growing type of treatments for wide range of diseases, with enormous potential for personalised medicine. Preparations of biopharmaceuticals for injections into patients should be free of contaminating endotoxin (Lipopolysaccharides, LPS), which otherwise may cause unwanted pyrogenic response and in its more severe form could lead to septic shock and death. LPS is a large amphiphilic molecule, comprised of a fatty acid component (Lipid A) and a polysaccharide component, containing phosphates and also glucosamine. This leads to hydrophilic 'outer component', an ionic core, and a hydrophilic 'inner' component; that means it can form micelles above a critical concentration (critical micelle concentration, CMC). LLPS can potentially interact with formulation components such as ions, buffers, and biopharmaceuticals themselves, which may complicate their detection and subsequent removal. The nature of specific and non-specific interactions that endotoxin monomers and micelles can have with different components of the formulation is as yet unstudied. The proposed project is aimed at developing novel approaches for monitoring behaviour of LLPS molecules in solution, and characterising their interactions with divalent cations, chelating agents, buffers and monoclonal antibodies, and exploring the way the clusters of these molecules are formed. The findings of this project will ultimately contribute to improving speed of bringing new and safer treatments to market and lowering costs of life saving drugs. The project will provide extensive training in advanced solution NMR techniques, LLPS chemistry and other biophysical methods, such as CG-MALS, DLS, and A4F, to obtain a comprehensive picture of inter- and intra-molecular interactions in this system. At least one month each year will be spent at placement at AstraZeneca's site in Cambridge, to run additional experiments and obtain further experience working in Industry.