SBIR Phase II: Synthetic biology platform for production of stabilized high-value proteins

这个小型企业创新研究(SBIR)第二阶段项目的更广泛影响是帮助糖尿病患者。这种疾病占美国死亡人数的12%，患者面临着重大的生活方式变化。大多数患者过渡到胰岛素替代疗法，这种疗法有一个复杂的剂量计划，如果不密切遵循，可能会使患者处于血糖失调的危险状态。目前有5000多万糖尿病患者使用基础胰岛素类似物，旨在比人类胰岛素活动更长时间。这些类比的便利性和更高的安全性已导致广泛采用，全球市场规模超过100亿美元。然而，目前所有的基础胰岛素都需要每天注射，这是一种剂量负担，导致治疗依从性差，使患者容易受到血糖危险波动的影响。此二期项目中描述的改良胰岛素旨在提供实现每周一次给药所需的稳定性。放宽注射时间表应该会极大地提高患者的依从性和安全性；此外，解决方案可以以更低的成本提供。该项目使用可扩展的体内蛋白质生产平台为糖尿病市场生产长效胰岛素类似物。该项目利用该平台的独特能力，将非标准氨基酸定点安装到蛋白质中，并规模化生产修饰蛋白质。通过用形成更强键的非标准氨基酸取代胰岛素中关键的成键氨基酸，修饰后的胰岛素可以实现支持每周一次给药所需的稳定性。研究目标是：生产足够数量的这种胰岛素类似物的变种以支持实验计划，在基于细胞的分析中证明变种比野生型胰岛素的稳定性更好，并在动物试验中证明胰岛素类似物的药效学充分延长，以支持每周给药一次。潜在的结果包括第一个能够满足临床和商业对负担得起的、安全的胰岛素类似物的主要需求的胰岛素类似物，给药时间表放松。此外，这项工作提供了一种新型蛋白质生产平台的技术验证。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact of this Small Business Innovation Research (SBIR) Phase II project is to help patients living with diabetes. The disease accounts for 12% of deaths in the US and patients face major lifestyle changes. Most patients transition to insulin replacement therapy, which carries a complex dosing schedule that, if not followed closely, can leave patients in dangerous states of glucose dysregulation. More than 50 million diabetics currently use basal insulin analogs designed for longer activity than human insulin. The convenience and improved safety of these analogs has led to widespread adoption and a global market surpassing $10B. However, all current basal insulins require daily injections, a dosing burden that leads to poor treatment adherence, leaving patients vulnerable to dangerous fluctuations in blood glucose. The modified insulin described in this Phase II project is intended to provide the stability necessary to achieve once-weekly dosing. Relaxing the injection schedule should dramatically improve compliance and safety for patients; furthermore, the solution can be delivered at lower cost. The project uses a scalable in vivo protein production platform to produce long-acting insulin analogs for the diabetes market. The project utilizes the platform’s unique capability to site-specifically install non-standard amino acids into proteins, and to produce the modified proteins at scale. By replacing key bond-forming amino acids in insulin with non-standard amino acids that form stronger bonds, the modified insulins can achieve the stability necessary to support once-weekly dosing. The research objectives are to: produce sufficient quantities of variants of this insulin analog to support an experimental program, demonstrate improved stability of the variants over wild-type insulin in cell-based assays, and demonstrate sufficiently prolongated pharmacodynamics of the insulin analogs in an animal study to support once-weekly dosing. Potential outcomes include the first insulin analog capable of filling a major clinical and commercial need for affordable, safe insulin analogs with relaxed dosing schedules. Further, the work provides technical validation of a novel protein production platform.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.