Engineering cells for concurrent protein drug biosynthesis and polysialylation

项目概要/摘要 治疗性蛋白质或生物制剂代表着一个价值 1000 亿美元的市场，其中包括药物 例如抗体、激素等等。生物制剂的临床疗效至关重要 由它们的循环半衰期决定。因此，人们开发了各种方法来 通过降低清除率来延长其循环半衰期。这通常是通过以下方式实现的 在体外将生物制品与生物相容性聚合物进行化学结合。然而，化学 缀合昂贵、复杂，并且常常导致特定活性的大量损失 以及异质产品混合物。这些严重的缺陷催生了需求 一项无需体外化学即可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，GlycoBac 提出了一种添加聚唾液酸的创新方法 生物制剂的生物合成过程中。聚唾液酸（PSA）天然存在于人体中， 是一种完全生物相容、可生物降解且非免疫原性的聚合物。体外化学 与相比，聚唾液酸化生物制剂已显示出更好的耐受性和药代动力学 为母药。此外，半个多世纪以来，唾液酸生物学已被充分了解。 研究。因此，PSA 是添加到生物制剂中的绝佳选择，目的是提高其活性 半衰期。我们的新方法使用糖蛋白生物制剂上现有的 N-聚糖作为支架 PSA 添加。用于生物生产的细胞已经将 N-聚糖添加到明确的位置。 我们建议在生物制剂过程中将 PSA 酶促添加到这些预先存在的 N-聚糖中。 生物合成（体内）。与化学缀合相反，我们的方法是位点特异性的，不 需要额外的处理步骤，并且不会引入额外的成本和复杂性。 该 SBIR 项目旨在证明体内聚唾液酸化作为下一步的可行性 生成平台技术。我们将通过专注于生产原型来实现这一目标 具有多唾液酸化途径的细胞系。这些细胞将用于产生两种聚唾液酸化的， 商业相关的糖蛋白生物制剂。对于第一阶段，我们将使用糖工程昆虫 细胞，因为 GlycoBac 在这种细胞类型方面拥有丰富的经验。我们的聚唾液酸化技术是 还与常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更大的第二阶段项目奠定基础，该项目的重点是 证明体内聚唾液酸化生物制剂的药代动力学和活性。第三阶段 我们与私营部门合作伙伴的体内聚唾液酸化技术的商业化是 预计将通过生产更有效的药物来显着影响人类健康 需要较少给药频率和/或减少剂量的糖蛋白生物制剂。

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.