Development of a novel production platform for complex recombinant biotherapeutic

开发复杂重组生物治疗的新型生产平台

• 批准号: 8251701
• 负责人: Gabriela Denning
• 金额: $ 29.63万
• 依托单位: EXPRESSION THERAPEUTICS
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8251701
• 关键词: A Mouse; Address; Adolescence; Amino Acids; Biochemical; Biological Products; Biological Response Modifier Therapy; Biomanufacturing; Biomedical Engineering; Bioreactors; Blood Coagulation Factor; Blood Proteins; Capital; Cell Aggregation; Cell Culture Techniques; Cell Density; Cell Line; Cells; Characteristics; Chinese Hamster Ovary Cell; Clinical; Column Chromatography; Complex; Conditioned Culture Media; Consumption; Development; Disease; Drug Formulations; Electroporation; Eukaryota; Evaluation; Factor IX; Factor VIII; Family suidae; Generations; Genetic; Goals; Gold; Growth; Hamsters; Harvest; Hemophilia A; Hormones; Human; Interferons; Ion Exchange; Kidney; Kilogram; Legal patent; Lentivirus Vector; Liposomes; M cell; Mammalian Cell; Marketing; Measures; Mediating; Methods; Modeling; Modification; Monoclonal Antibodies; Nutrient; Persons; Pharmacologic Substance; Pharmacy facility; Phase; Polymers; Post-Translational Protein Processing; Price; Procedures; Production; Productivity; Prokaryotic Cells; Protein Biosynthesis; Proteins; Recombinant Proteins; Recombinants; Relative (related person); Roller Bottle; Running; Serum; Serum Proteins; Serum-Free Culture Media; Small Business Innovation Research Grant; Suspension Culture; Suspension substance; Suspensions; System; Technology; Testing; Therapeutic; Thrombin; Time; Validation; Yeasts; base; blood product; cost; cytokine; design; flasks; flexibility; genetically modified cells; improved; in vivo; kidney cell; manufacturing process; mortality; novel; pre-clinical; recombinant antihemophilic factor VIII; research and development

DESCRIPTION (provided by applicant): The manufacturing of recombinant protein-based biopharmaceuticals is a complex, labor and capital intensive endeavor. It currently is accepted that the use of mammalian cells is a requirement for the production of most human proteins, which typically contain extensive post-translational modifications that are not performed by prokaryotes and single-celled eukaryotes. Although mammalian cells such as Chinese hamster ovary cells and baby hamster kidney cells can faithfully biosynthesize most human proteins, the efficiency is dramatically lower than is achieved by bacterial or yeast cells. Of the recombinant proteins currently marketed, coagulation factor VIII is manufactured with the lowest efficiency and is by far the most expensive on a per unit mass basis. Other expensive and difficult to manufacture biopharmaceuticals include the other coagulation factors (e.g. factors IX and VIIa), interferon, and hormone-based cytokines. The goal of the current application is to develop a technology that can improve the production volume and pharmacy price of these difficult to manufacture recombinant biologics. Expression Therapeutics has developed a new baby hamster kidney-derived (BHK) cell line, designated BHK-MS, which can be cultured and expanded indefinitely in serum and blood product-free medium under suspension bioreactor conditions. In the current studies, we are 1) developing and validating the suspension adapted BHK-MS cell line for superior coagulation factor VIII and factor IX production, and 2) performing a pilot-scale manufacturing run and analyzing the biochemical and pharmaceutical characteristics of BHK-MS biosynthesized recombinant factor VIII and factor IX, which will serve as a model, difficult to manufacture biopharmaceuticals. PUBLIC HEALTH RELEVANCE: The long-term commercial objective of the current project is to commercialize a suspension cell-based biomanufacturing platform designed for improved volumetric productivity of complex, hard to manufacture biologics. Low-yield manufacturing processes currently in place often result in limited product availability and high-product costs. Expression Therapeutics has developed a patent-pending cell line designed for efficient manufacturing of recombinant biologics. The studies proposed herein are designed to provide proof-of-concept of the efficiency and scalability, as well as, rigorous preclinical validation and characterization of this novel manufacturing platform.

描述（由申请人提供）：重组蛋白生物制药的生产是一项复杂的劳动和资本密集型工作。目前公认的是，使用哺乳动物细胞是生产大多数人类蛋白质的必要条件，这些蛋白质通常含有原核生物和单细胞真核生物无法进行的广泛的翻译后修饰。尽管哺乳动物细胞如中国仓鼠卵巢细胞和小仓鼠肾细胞可以真实地生物合成大多数人类蛋白质，但其效率明显低于细菌或酵母细胞。在目前上市的重组蛋白中，凝血因子VIII的生产效率最低，单位质量的成本也是迄今为止最高的。其他昂贵且难以制造的生物药品包括其他凝血因子（如因子IX和因子VIIa）、干扰素和基于激素的细胞因子。目前申请的目标是开发一种技术，可以提高这些难以生产的重组生物制剂的产量和药品价格。Expression Therapeutics开发了一种新的幼鼠肾源性（BHK）细胞系，命名为BHK- ms，可以在悬浮生物反应器条件下在血清和无血液制品的培养基中无限培养和扩增。在目前的研究中，我们正在1)开发和验证适用于BHK-MS细胞系的悬浮液，用于更好的凝血因子VIII和因子IX的生产；2)进行中试生产，并分析BHK-MS生物合成重组因子VIII和因子IX的生化和药物特性，这将作为一个模型，难以制造生物制药。