Development of predictive tools and formulations for improved stability and delivery

开发预测工具和配方以提高稳定性和交付

• 批准号: 1667876
• 金额: --
• 依托单位: University of Kent
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1667876
• 关键词: Development; predictive; tools; formulations; improved

Project Description:Industry traditionally develops biotherapeutic recombinant protein and peptide formulations to stabilise and deliver such drugs by undertaking pre-formulation studies that identify drug concentration, excipient, buffer and other formulation conditions that provide a formulation fit for purpose (i.e. one that has the required stability and can be delivered in an active form without degradation of the bioactive drug). Traditionally the development of such formulations is undertaken using design of experiment approaches to screen a large number of potential variables and 'identify' a 'suitable' formulation for a specific product. This approach generally works well for established biotherapeutics, for example monoclonal antibodies (mAbs), where historical knowledge is used as a starting point in the development of a new molecule, although even in this field for some molecules it can be difficult to stabilise the molecule at the desired concentration. For non-mAb biotherapeutics it can be more challenging to design appropriate formulations due to the divergent nature of the molecules, lack of historical data on these and our general limited understanding of the relationship between stability, recombinant proteins/peptides and formulations. In this project we will investigate two classes of non-mAb based biotherapeutics, peptides and heavy chain fusion proteins, and the influence of residue host cell proteins and excipient conditions on subsequent stability using accelerated and long term stability studies. The student will express in CHO or E. coli cells model peptide and heavy chain fusion proteins (milestone 1) and then purify these using affinity and ion exchange chromatography (milestone 2). The student will use mass spectrometry, ELISA and gel based approaches throughout the purification process to determine the amounts of host cell protein remaining and relate this to subsequent stability of the molecule under standard accelerated stability studies (as required for regulator approval of final products) monitored by DSC, CD, particle formation analysis, size exclusion chromatography, cell based activity assays (where appropriate) and mass spectrometry (milestone 3). The purified materials will then be subjected to design of experiments analysis of formulation variables at MedImmune using standard industrial procedures to develop and test formulations for biotherapeutic products (milestone 4). In silico analysis will also be undertaken to highlight potential amino acids or areas that might introduce instability into the molecules and site directed mutants of these produced and related to stability using the design-of-experiments formulation screen (milestone 5)The proposed programme of work will improve our understanding of the molecular mechanisms by which proteins may be stabilised. We also note that trained formulation scientists in biotherapeutics (as opposed to small molecules) are difficult to recruit in the UK and that this is an area of Bioprocessing. This project will train a formulation scientist and further our understanding of the issues preventing stable formulation of a number of key therapeutic target molecules. This project arises as a direct result of a successful collaborative PhD studentship between the two institutes in formulation studies.

项目简介：业界传统上开发生物治疗性重组蛋白质和多肽制剂，通过进行配方前研究来确定药物浓度、赋形剂、缓冲剂和其他配方条件，以提供适合用途的配方(即具有所需稳定性并可以以活性形式输送而不会降解生物活性药物的制剂)。传统上，此类配方的开发是通过设计实验方法来筛选大量潜在变量，并为特定产品‘确定’一个‘合适’的配方。这种方法通常适用于已有的生物疗法，例如单抗(MAbs)，其中历史知识被用作开发新分子的起点，尽管即使在这一领域，对于某些分子来说，也很难将分子稳定在所需的浓度。对于非单抗生物疗法，由于分子的不同性质，缺乏这些方面的历史数据，以及我们对稳定性、重组蛋白质/肽和制剂之间的关系的一般有限的理解，设计适当的制剂可能更具挑战性。在这个项目中，我们将通过加速和长期稳定性研究来考察两类基于非单抗的生物治疗药物，多肽和重链融合蛋白，以及残存的宿主细胞蛋白和赋形剂条件对后续稳定性的影响。学生将在CHO或E.Coli细胞模型中表达多肽和重链融合蛋白(里程碑1)，然后使用亲和和离子交换层析(里程碑2)纯化它们。学生将在整个纯化过程中使用质谱仪、ELISA和基于凝胶的方法来确定宿主细胞剩余蛋白质的量，并将其与DSC、CD、颗粒形成分析、尺寸排除层析、基于细胞的活性分析(适当时)和质谱仪(里程碑3)监测的标准加速稳定性研究(监管机构对最终产品的批准所需)下随后的分子稳定性联系起来。然后，提纯后的材料将在MedImmune接受实验设计和配方变量分析，使用标准工业程序开发和测试生物治疗产品的配方(里程碑4)。在计算机分析中，还将利用实验设计配方筛选(里程碑5)来突出可能给分子带来不稳定的潜在氨基酸或区域，以及这些氨基酸的定点突变和与稳定性相关的突变。拟议的工作计划将提高我们对蛋白质可能稳定的分子机制的理解。我们还注意到，在英国很难招募到训练有素的生物疗法(而不是小分子)配方科学家，这是生物加工领域的一个领域。这个项目将培训一名配方科学家，并进一步加深我们对阻碍一些关键治疗靶标分子稳定配方的问题的理解。这个项目是两个研究所在配方研究方面成功合作的博士生的直接结果。