Enabling implantable artificial pancreas pumps with heat-stable, ultra-rapid insulin

项目摘要 我们寻求开发一种超稳定超快速胰岛素类似物配方，用于先进的“智能”泵， 糖尿病的治疗。一种市售胰岛素制剂，在体内稳定， 在室温下至少180天和在室温下一年将分别使（a） 用于治疗T1DM/T2DM的植入式腹膜内闭环泵和（B）预填充贴片 治疗T2DM的胰岛素泵。后一种市场的经济学使发展一种 用于连接到连续葡萄糖的植入式人工胰腺（"AP"）装置的关键使能技术 监视器.超快速药代动力学/动力学（PK/PD）有望提高药物的安全性和有效性。 在这种闭环系统中采用的反馈算法。 目前开发更稳定的胰岛素制剂的障碍是温度依赖性 胰岛素对纤维化的敏感性;这种物理降解导致促炎性淀粉样蛋白。 在体温条件下，商业胰岛素制剂中的原纤维可以在温和的一周内形成 搅拌（如在泵储液器中）。一旦纤维化开始，一个种子成核生长过程促进了纤维化。 天然胰岛素分子迅速转化为淀粉样蛋白;活性呈指数下降，使剂量 不准确并留下导致导管堵塞的沉积物。 为了克服这一障碍，提出了一种基于单链胰岛素的创新结构方法 (SCI)平台是完全有效的，但对原纤化和化学降解是难治的。设计这种 平台基于（i）最近关于胰岛素如何与其主要结合位点相互作用的晶体学研究， 胰岛素受体（"位点1"）和（ii）胰岛素原纤维的分子模型。特别是，我们发现， 在B链（ThrB 30）的C-末端和A链（ThrB 30）的N-末端之间适当构建的6 - 8个残基接头 链（GlyA1）可以防止原纤维形成> 1年的温和搅拌在37 ℃，同时保持天然的 生物活性这样的SCI在无锌单体制剂和锌基制剂中都是稳定的。 六聚体制剂，从而在选择用于同时给药的赋形剂方面提供显著的灵活性。 优化稳定性和吸收速率（快速启动）。我们将把这种优化扩展到快速工程- 通过修饰胰岛素的辅助受体结合表面（与 受体α-亚基纤连蛋白同源结构域中的位点2。 超稳定的快速开/关SCI制剂将提供AP技术的重大进步。我们 因此，建议合成并表征五种此类SCIs作为候选制剂。B.H.博士弗兰克 （主要研究者）在Eli Lilly之前的职业生涯中是Humidiform的共同发明人。Thermalin糖尿病， LLC拥有SCI相关知识产权的独家许可，该知识产权归CWRU所有。

Project Summary We seek to develop an ultra-stable ultra-fast insulin analog formulation for use in advanced “smart” pumps in the treatment of diabetes mellitus. A commercially available insulin formulation that is stable at body temperature for at least 180 days and at room temperature for a year would respectively make practical (a) implantable intraperitoneal closed-loop pumps for the treatment of T1DM/T2DM and (b) pre-filled patch pumps for the treatment of T2DM. The economics of the latter market makes practical the development of a critical enabling technology for an implanted artificial pancreas (“AP”) device linked to a continuous glucose monitor. Ultra-fast pharmacokinetic/dynamic (PK/PD) promises to improve the safety and efficacy of the feedback algorithms employed in such closed-loop systems. The current barrier to the development of more stable insulin formulations is the temperature-dependent susceptibility of insulin to undergo fibrillation; such physical degradation leads to a pro-inflammatory amyloid. At body temperature fibrils can form in commercial insulin formulations in as little as one week on gentle agitation (as in a pump reservoir). Once fibrillation begins, a seeded nucleation-growth process promotes the rapid conversion of the native insulin molecules into amyloid; activity declines exponentially, making dosing inaccurate and leaving deposits that lead to catheter occlusion. To overcome this barrier, an innovative structural approach is proposed based on a single-chain insulin (SCI) platform that is fully potent and yet refractory to fibrillation and chemical degradation. Design of this platform is based on (i) recent crystallographic studies of how insulin interacts with its primary binding site in the insulin receptor (“Site 1”) and (ii) molecular models of insulin fibrils. In particular, we have discovered that a properly constructed 6-8 residue linker between the C-terminus of the B chain (ThrB30) and N-terminus of the A chain (GlyA1) can prevent fibril formation for >1 year on gentle agitation at 37 oC while preserving native biological activity. Such an SCI is stable both in a zinc-free monomeric formulation and in a zinc-based hexameric formulation, thus providing marked flexibility in choice of excipients for the simultaneous optimization of stability and rate of absorption (fast-ON). We will extend such optimization to engineer fast- OFF pharmacodynamics (PD) through modification of insulin’s ancillary receptor-binding surface, cognate to Site 2 in the fibronectin-homology domains of the receptor -subunit. An ultra-stable fast-ON/fast-OFF SCI formulation would provide a major advance in AP technology. We therefore propose to synthesize and characterize five such SCIs as candidate formulations. Dr. B.H. Frank (principal investigator) was co-inventor of Humalog during his prior career at Eli Lilly. Thermalin Diabetes, LLC has an exclusive license to SCI-related IP, which is owned by CWRU.