Engineering cells for concurrent protein drug biosynthesis and polysialylation

用于并行蛋白质药物生物合成和聚唾液酸化的工程细胞

• 批准号: 8645308
• 负责人: Christoph Geisler
• 金额: $ 14.82万
• 依托单位: GLYCOBAC, LLC
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8645308
• 关键词: Address; Anabolism; Antibodies; Baculoviruses; Biocompatible; Biology; Cell Line; Cells; Chemicals; Chemistry; Data; Dose; Drug Kinetics; Erythropoietin; Escherichia coli; Future; Glycoproteins; Goals; Growth; Half-Life; Health; Heterogeneity; Hormones; Human; Human body; In Vitro; Insecta; Intellectual Property; Length; Letters; Licensing; Mammalian Cell; Manufacturer Name; Marketing; Measures; Medicine; Metabolic Clearance Rate; Methods; Nature; Parents; Pathway interactions; Pharmaceutical Preparations; Phase; Polymers; Polysaccharides; Polysialic Acid; Positioning Attribute; Private Sector; Process; Production; Productivity; Proteins; Qualifying; Recombinant Proteins; Recombinants; Research; Sialic Acids; Site; Small Business Innovation Research Grant; Staging; Technology; Testing; Therapeutic; Universities; Wyoming; alpha 1-Antitrypsin; biocompatible polymer; biodegradable polymer; cell type; cellular engineering; clinical efficacy; commercialization; cost; design; dosage; experience; glycosyltransferase; improved; in vivo; innovation; meetings; next generation; prototype; public health relevance; research and development; scaffold; success; therapeutic protein

Project Summary / Abstract Therapeutic proteins, or biologics, represent a $100 billion market that includes drugs such as antibodies, hormones, and many others. The clinical efficacy of biologics is critically determined by their circulating half-lives. Hence, various methods have been developed to increase their circulating half-lives by reducing clearance rates. This is commonly achieved by chemically conjugating biologics with biocompatible polymers in vitro. However, chemical conjugation is expensive, complicated, and often results in substantial losses of specific activity as well as a heterogeneous product mixture. These serious drawbacks have created a demand for a technology that can add biocompatible polymers to biologics without in vitro chemistry. To meet this demand, GlycoBac proposes a new, innovative method to add polysialic acid to biologics during their biosynthesis. Polysialic acid is (PSA) naturally found in the human body, and is a fully biocompatible, biodegradable and non-immunogenic polymer. In vitro chemically polysialylated biologics have already shown improved tolerance and pharmacokinetics compared to parent drugs. Moreover, sialic acid biology is well-understood through over half a century of research. Thus, PSA is an excellent choice to add to biologics with the goal of increasing their half-lives. Our new method uses existing N-glycans on glycoprotein biologics as a scaffold for PSA addition. Cells used for biologic production already add N-glycans to well-defined positions. We propose to enzymatically add PSA to these pre-existing N-glycans during biologic biosynthesis (in vivo). In contrast to chemical conjugation, our method is site-specific, does not require additional processing steps, and does not introduce additional cost and complexity. This SBIR project is designed to prove the feasibility of in vivo polysialylation as a next- generation platform technology. We will achieve this by Aims focused on producing a prototype cell line with a polysialylation pathway. These cells will be used to produce two polysialylated, commercially relevant glycoprotein biologics. For Phase I, we will use glycoengineered insect cells, as GlycoBac has extensive experience with this cell type. Our polysialylation technology is also compatible with mammalian cell lines such as CHO and PerC.6, which are commonly used to produce biologics. Phase I success will set the stage for a larger Phase II project focused on demonstrating the pharmacokinetics and activity of in vivo polysialylated biologics. Phase III commercialization of our in vivo polysialylation technology with private-sector partners is expected to significantly impact human health by enabling production of more efficacious glycoprotein biologics that require less-frequent dosing and/or reduced dosages.

项目摘要 /摘要 治疗蛋白或生物制剂代表了1000亿美元的市场，包括药物 例如抗体，激素和许多其他。生物制剂的临床功效至关重要 取决于他们循环的半衰期。因此，已经开发了各种方法 通过降低清除率来增加循环的半衰期。这通常是由 在体外与生物相容性聚合物化学结合生物制剂。但是，化学 共轭昂贵，复杂，并且通常会导致特定活动的大量损失 以及异质产品混合物。这些严重的缺点创造了需求 对于可以在没有体外化学的情况下将生物相容性聚合物添加到生物制剂中的技术。 为了满足这一需求，糖果实提出了一种新的创新方法来添加多氨基酸 在生物合成期间生物制剂。多胺酸是自然发现的（PSA），在人体中， 并且是一种完全具有生物相容性的，可生物降解和非免疫原性聚合物。化学上体外 多酰化生物制剂已经显示出提高的耐受性和药代动力学比较 给家长毒品。此外，在半个多世纪的 研究。因此，PSA是增加生物制剂的绝佳选择，目的是增加其 半衰期。我们的新方法在糖蛋白生物制剂上使用现有的N-聚糖作为脚手 PSA添加。用于生物生产的细胞已经在定义明确的位置中添加了N-聚糖。 我们建议在生物学期间酶促添加PSA 生物合成（体内）。与化学共轭相反，我们的方法是特定于地点的，不是 需要其他处理步骤，并且不引入额外的成本和复杂性。 该SBIR项目旨在证明体内多酰基化作为下一步的可行性 生成平台技术。我们将通过专注于制作原型的目标来实现这一目标 带有多准会途径的细胞系。这些细胞将用于产生两个多酶促的， 商业相关的糖蛋白生物制剂。对于第一阶段，我们将使用糖化昆虫 细胞，因为糖果在这种细胞类型方面具有丰富的经验。我们的多酰基化技术是 也与通常使用的哺乳动物细胞系（例如Cho和Perc.6）兼容 生产生物制剂。第一阶段的成功将为一个重点的II阶段项目奠定基础 证明体内多糖化生物制剂的药代动力学和活性。第三阶段 与私营部门合作伙伴对我们体内多准会技术的商业化是 预计通过产生更有效的生产会对人类健康产生重大影响 糖蛋白生物制剂需要少量给药和/或降低剂量。