Manipulating Aromaticity: characterization of an ultra-rapid insulin analog

控制芳香度：超快速胰岛素类似物的表征

• 批准号: 8511621
• 负责人: Bruce Hill Frank
• 金额: $ 26.35万
• 依托单位: THERMALIN DIABETES, LLC
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-16 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8511621
• 关键词: Algorithms; Amino Acids; Artificial Pancreas; Blood; Blood Glucose; Boxing; Budgets; Buffers; Clinical; Clinical Research; Clinical Trials; Color; Cystine; Development; Diabetes Mellitus; Digestion; Dose; Drug Formulations; Drug Kinetics; Enzyme-Linked Immunosorbent Assay; Event; Family suidae; Feedback; Generations; Glycosylated hemoglobin A; Gold; Gray unit of radiation dose; Hyperglycemia; Hypoglycemia; Infusion procedures; Injection of therapeutic agent; Insulin; Insulin Infusion Systems; Insulin, Aspart, Human; Insulin, Lispro, Human; Insulin-Dependent Diabetes Mellitus; Ions; Kinetics; Lead; Left; Letters; Licensing; Life; Light; Mediating; Modeling; Mutagenesis; Mutation; NPM1 gene; National Institute of Diabetes and Digestive and Kidney Diseases; NovoLog; Octreotide; Parents; Partner in relationship; Patients; Performance; Pharmacodynamics; Phase; Physical condensation; Positioning Attribute; Principal Investigator; Production; Proteins; Protomer; Pump; Rattus; Regimen; Relative (related person); Risk; Safety; Sampling; Series; Serum; Side; Site-Directed Mutagenesis; Small Business Innovation Research Grant; Solubility; Stream; Streptozocin; Structure; Study Section; Subcutaneous Injections; Suggestion; Sulfur; System; Technology; Testing; Thermodynamics; Time; TimeLine; Trypsin; Zinc; absorption; analog; animal data; base; career; chemical stability; chemical synthesis; design; dimer; glucose monitor; glycemic control; improved; in vivo; innovation; insulin dimers; non-diabetic; pre-clinical; programs; response; subcutaneous

DESCRIPTION (provided by applicant): We seek to develop an ultra-fast insulin analog formulation for the treatment of diabetes mellitus. Ultra-fast pharmacokinetic/dynamic (PK/PD) promises to enable superior performance of pump therapy (continuous subcutaneous insulin infusion; CSII) with enhanced safety and more robust integration with continuous glucose monitors (CGM). Our product promises to improve the safety and efficacy of algorithm-controlled closed-loop systems (the "artificial pancreas"); ultra-fast PK/PD would also facilitate post-prandial glycemic control in conventional multi-injection regimens. An innovative structural approach is proposed based on non-standard mutagenesis of the insulin molecule. This application has been revised based on the suggestions of the Study Section, most saliently to enhance its focus on stability and to provide supporting animal data. The current barrier to more rapid subcutaneous absorption is the rate of disassembly of the zinc insulin hexamer. This rate is controlled by the stability of an anti-parallel ¿-sheet at the dimer interface of the hexamr. Partial progress toward accelerated disassembly following subcutaneous injection was achieved in the 1990s by conventional site-directed mutagenesis at the periphery of this ¿-sheet. Insulin aspart (the active component of Novolog(R); Novo-Nordisk) and insulin lispro (the active component of Humalog(R); Eli Lilly) contain such partially destabilizing mutations (ProB28->Asp in Novolog; ProB28->Lys and LysB29->Pro in Humalog ("KP")). Because their designs were limited by conventional mutagenesis, however, neither of these products could fully exploit structural strategies to promote disassembly of the core ¿-sheet in the insulin hexamer. The core ¿ -sheet of the insulin hexamer is remarkable for the confluence of 8 conserved aromatic rings (TyrB16, PheB24, PheB25, TyrB26, and their dimer-related mates). The thermodynamic and kinetic stability of this interface is regulated by an intricate series of aromatic-aromatic interactions. We have discovered that the substitution of PheB24 by its non-aromatic analog cyclohexanylalanine (Cha) is compatible with native-like structure and function, but markedly enhances the rate of disassembly of KP-insulin. Since submission of the original application, we have also invented an ultra-stable version of this analog containing additional A-chain substitution ThrA8->Glu. We therefore propose to synthesize and characterize ChaB24-KP-insulin and ChaB24- GluA8-insulin as candidate ultra-rapid formulations. Dr. B. Frank (principal investigator) was co-inventor of Humalog(R) during his prior career at Eli Lilly. Thermalin Diabetes, LLC has an exclusive license to ChaB24- related IP, which is owned by CWRU. We thank the Study Section for its guidance.

描述（由应用提供）：我们试图开发一种用于治疗糖尿病的超快速胰岛素模拟公式。超快速的药代动力学/动力学（PK/PD）有望使泵治疗（连续的皮下胰岛素输注； CSII）具有卓越的性能，并具有增强的安全性和与连续葡萄糖监测仪（CGM）更强的结合。我们的产品有望提高算法控制的闭环系统的安全性和效率（“人造胰腺”）；超快速的PK/PD还将促进常规多注射方案中的后原始血糖控制。基于胰岛素分子的非标准诱变提出了一种创新的结构方法。根据研究部分的建议，对该应用程序进行了修订，最明显地提高了其对稳定性的关注并提供支持动物数据。当前皮下吸收更快的障碍是锌胰岛素己酯的拆卸速率。该速率由六二聚体界面处的反平行� -sheet的稳定性控制。在1990年代，通过常规位置定向的诱变在此€-sheet的外围，在1990年代采取了皮下注射后加速拆卸的部分进展。胰岛素阿斯帕特（Novolog（R）的活性组成部分； Novo-Nordisk）和胰岛素lispro（HumanLog（R）的活性成分（R）； Eli Lilly）包含这种部分破坏性的突变（Novolog in Novolog; Prob28-> lys和Lysb29-> pro in HumanLog In HumanLog（nov28-> lys28-> lys28-> lys））。但是，由于它们的设计受到常规诱变的限制，因此，这些产品都无法完全利用结构策略来促进胰岛素己酸酯中核心表的拆卸。胰岛素六聚体的核心表对于8个保守的芳环（Tyrb16，Pheb24，Pheb25，Pheb25，Tyrb26及其与二聚体相关的伴侣）而言是显着的。该界面的热力学和动力学稳定性受一系列复杂的芳香芳香族相互作用的调节。我们已经发现，Pheb24的非芳基类似环己基丙氨酸（CHA）与天然的结构和功能兼容，但显着提高了KP-胰岛素的拆卸率。自从提交原始应用程序以来，我们还发明了此模拟的超稳定版本，其中包含其他A链替代thra8-> glu。因此，我们建议合成并表征CHAB24-KP-胰岛素和CHAB24-GLUA8-胰岛素作为候选超优化公式。 B. Frank博士（首席研究员）在Eli Lilly的职业生涯中是HumanLog（R）的共同发明。 Thermalin Diabetes，LLC拥有CWRU拥有的CHAB24相关IP的独家许可。我们感谢研究部分的指导。