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

这项小型企业创新研究（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改变。 大多数患者过渡到胰岛素替代疗法，其具有复杂的给药方案，如果不密切遵循，可能使患者处于葡萄糖调节异常的危险状态。目前有超过5000万糖尿病患者使用基础胰岛素类似物，其活性比人胰岛素更长。这些类似物的便利性和更高的安全性导致了广泛的采用和超过100亿美元的全球市场。然而，目前所有的基础胰岛素都需要每天注射，这是一种给药负担，导致治疗依从性差，使患者容易受到血糖危险波动的影响。本II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。放松注射时间表应该大大提高患者的依从性和安全性;此外，该解决方案可以以较低的成本提供。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，并大规模生产改性蛋白质。通过用形成更强键的非标准氨基酸取代胰岛素中的关键成键氨基酸，修饰的胰岛素可以实现支持每周一次给药所需的稳定性。研究目的是：生产足够量的这种胰岛素类似物的变体以支持实验计划，在基于细胞的测定中证明变体相对于野生型胰岛素的稳定性改善，并在动物研究中证明胰岛素类似物的充分增强的药效学以支持每周一次给药。潜在的成果包括第一个胰岛素类似物能够满足主要的临床和商业需求，负担得起的，安全的胰岛素类似物与宽松的给药方案。此外，这项工作提供了一个新的蛋白质生产平台的技术验证。这个奖项反映了NSF的法定使命，并已被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。

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.