Optimized Receptor Binding Profile in an Ultra-Stable, Ultra-Rapid-Acting Insulin

描述（由申请人提供）：我们寻求开发一种用于治疗糖尿病的超快速无锌胰岛素类似物配方。超快速药代动力学（PK/PD）分析有望实现泵治疗（持续皮下胰岛素输注）的卓越性能，提高安全性，并与葡萄糖传感技术更强大的集成。超快速动力学也有助于单次餐后注射后的餐后血糖控制。皮下吸收的障碍是胰岛素的自组装。因此，超快速配方的关键组成部分将是一种具有足够内在化学和物理稳定性的工程胰岛素单体，使锌介导的自组装变得不必要。这种类似物早在20年前就被开发出来（aspb10 -胰岛素），但由于其潜在的致瘤性和相对于天然胰岛素的体外有丝分裂性增加，未能进行临床前测试。这些特性被认为反映了与igf -1受体（IGF-1R）结合增加和在胰岛素受体（IR）停留时间延长。我们发现，AspB10-胰岛素类似物的战略性氟修饰(a)消除了与IGF-1R的不良结合和延长的IR停留时间，同时(b)增强了AspB10的稳定作用。氟修饰残基为邻位f - pheb24，可通过化学合成或新型基因工程插入。因此，需要i期支持，以实现与含有B28和B29位置速效b链取代的含氟AspB10类似物的稳定性、效力、有丝分裂性和PK/PD相关的里程碑(源自当前产品Humalog（R)和Novolog(R)）。本建议创新性地利用氟（药物化学的支柱）在蛋白质生物技术中，以提高胰岛素替代疗法的安全性和有效性。

DESCRIPTION (provided by applicant): We seek to develop an ultra-fast, zinc-free insulin analog formulation for the treatment of diabetes mellitus. An ultra-fast pharmacokinetic-dynamic (PK/PD) profile promises to enable superior performance of pump therapy (continuous subcutaneous insulin infusion) with enhanced safety and more robust integration with glucose- sensing technologies. Ultra-fast kinetics would also facilitate post-prandial glycemic control following single mealtime injections. The barrier to subcutaneous absorption is insulin self-assembly. The key component of an ultra-fast formulation would, thus, be an engineered insulin monomer with sufficient intrinsic chemical and physical stability to render zinc-mediated self-assembly unnecessary. Such an analog was developed 20 years ago (AspB10-insulin), but failed preclinical testing due to its potential tumorigenicity and increased in vitro mitogenicity relative to native insulin. These properties are thought to reflect increased binding to the IGF-I receptor (IGF-1R) and prolonged residence time at the insulin receptor (IR). We have discovered that a strategic fluoro-modification of an AspB10-insulin analog (a) eliminates undesirable binding to IGF-1R and prolonged IR residence time and, at the same time, (b) enhances the stabilizing effects of AspB10. The fluoro-modified residue is ortho-F-PheB24, which is amenable to insertion by chemical synthesis or by novel genetic engineering. Phase-I support is, therefore, requested to achieve milestones related to the stability, potency, mitogenicity, and PK/PD of fluoro-protected AspB10 analogs containing rapid-acting B-chain substitutions at positions B28 and B29 (derived from current products Humalog(R) and Novolog(R)). This proposal makes innovative use of fluorine (a mainstay of medicinal chemistry) in protein biotechnology to enhance the safety and efficacy of insulin replacement therapy. PUBLIC HEALTH RELEVANCE: Diabetes is increasing in global prevalence. To provide greater convenience, improved glycemic control, and fewer adverse side-effects (all of which result in greater compliance and lower healthcare costs), we have invented novel ultra-stable and ultra-rapid-acting insulin analogs (designated Fluorolog-1 and Fluorolog-2) that exhibit optimized receptor binding profiles attenuating unwanted effects of AspB10 on binding to the insulin receptor (IR) and the IGF-I receptor (IGF-1R), in principle reducing cancer risk. The innovative design of these analogs exploits fluorine-based electrostatic engineering to "tune" the stability, mitogenicity, and potency of an engineered Zn-free insulin monomer. This project will complete feasibility testing on Fluorolog-1 and Fluorolog-2.