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增长最快的类型，具有巨大的个性化医疗潜力。用于注射到患者体内的生物药物制剂应不含污染性内毒素（脂多糖，LPS），否则可能导致不必要的热原反应，并且在其更严重的形式下可能导致败血性休克和死亡。LPS是一种大的两亲分子，由脂肪酸组分（脂质A）和多糖组分组成，含有磷酸盐和葡糖胺。这导致亲水性的“外部组分”，离子核心和亲水性的“内部”组分;这意味着它可以在临界浓度（临界胶束浓度，CMC）以上形成胶束。LLPS可能与制剂组分（如离子、缓冲液和生物药物本身）相互作用，这可能使其检测和随后的去除复杂化。内毒素单体和胶束与制剂不同组分的特异性和非特异性相互作用的性质尚未研究。 拟议的项目旨在开发新的方法来监测溶液中LLPS分子的行为，并表征它们与二价阳离子，螯合剂，缓冲液和单克隆抗体的相互作用，并探索这些分子形成簇的方式。该项目的研究结果最终将有助于加快将新的、更安全的治疗方法推向市场的速度，并降低挽救生命的药物的成本。 该项目将提供先进的溶液NMR技术，LLPS化学和其他生物物理方法（如CG-MALS，DLS和A4 F）的广泛培训，以获得该系统中分子间和分子内相互作用的全面图像。每年至少有一个月的时间将在阿斯利康剑桥的工厂实习，以进行额外的实验并获得更多的工业工作经验。

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.