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（由申请人提供）：我们寻求开发一种用于治疗糖尿病的超快速胰岛素类似物制剂，超快速药代动力学/动力学（PK/PD）有望实现泵疗法（连续皮下胰岛素输注； CSII）具有增强的安全性以及与连续血糖监测仪（CGM）更强大的集成，我们的产品有望提高算法控制的闭环系统（“人工胰腺”）的安全性和有效性； PK/PD 还将促进传统多次注射方案中的餐后血糖控制。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。该申请已根据研究部分的建议进行了修订。显着加强其对稳定性的关注并提供支持性动物数据，目前阻碍更快速皮下吸收的障碍是锌胰岛素六聚体的分解速率，该速率由反平行的稳定性控制。 20 世纪 90 年代，通过在 hexamr 的二聚体界面处进行传统的定点诱变，在皮下注射后加速分解方面取得了部分进展。 -门冬胰岛素（Novolog(R) 的活性成分；Novo-Nordisk）和赖脯胰岛素（Humalog(R) 的活性成分；Eli Lilly）含有此类部分不稳定突变（Novolog 中的 ProB28->Asp；ProB28-然而，由于它们的设计受到传统诱变的限制，因此它们都不是 Humalog 中的 >Lys 和 LysB29->Pro。产品可以充分利用结构策略来促进核心的拆卸 ¿ - 胰岛素六聚体的片层 ¿胰岛素六聚体的片层因 8 个保守芳香环（TyrB16、PheB24、PheB25、TyrB26 及其二聚体相关配偶体）的汇合而引人注目，该界面的热力学和动力学稳定性受到一系列复杂的芳香族-的调节。我们发现 PheB24 被其非芳香族类似物环己基丙氨酸 (Cha) 取代与天然类似物兼容。结构和功能，但显着提高了 KP-胰岛素的分解率，自提交原始申请以来，我们还发明了该类似物的超稳定版本，其中包含额外的 A 链替代 ThrA8->Glu。合成并表征了 ChaB24-KP-胰岛素和 ChaB24-GluA8-胰岛素作为候选超快速制剂。B. Frank 博士（首席研究员）是 的共同发明人。 Humalog(R) 在 Eli Lilly 的职业生涯中拥有 ChaB24 相关知识产权的独家许可，该许可由 CWRU 所有。