Molecular engineering of complementary glucose-responsive conformational switches in insulin and glucagon

胰岛素和胰高血糖素中互补葡萄糖响应构象开关的分子工程

• 批准号: 10263301
• 负责人: FARAMARZ ISMAIL-BEIGI
• 金额: $ 58.36万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10263301
• 关键词: 3-Dimensional; Affinity; Algorithms; Animal Testing; Animals; Attenuated; B-Lymphocytes; Binding; Biological Assay; Biophysics; Blood; Blood Glucose; Boron; Boronic Acids; Carbohydrates; Cell Culture Techniques; Cell physiology; Chemistry; Clinical; Clinical Endocrinology; Clinical Management; Closure by clamp; Complex; Computer Simulation; Computers; Cryoelectron Microscopy; Crystallization; Data; Dependence; Development; Devices; Diabetes Mellitus; Dose; Elements; Endocrinology; Engineering; Exclusion; Female; Formulation; Foundations; Glucagon; Glucagon Receptor; Glucose; Glycols; Goals; Health; Heteronuclear NMR; Homeostasis; Hormonal; Hormone Receptor; Hormone replacement therapy; Hormones; Hyperglycemia; Hypoglycemia; Injections; Insulin; Insulin Infusion Systems; Insulin Receptor; Insulin, Lispro, Human; Insulin-Dependent Diabetes Mellitus; Joints; Lead; Ligands; Measures; Metabolic; Metabolism; Methods; Modeling; Molecular; Molecular Conformation; Nature; Non-Insulin-Dependent Diabetes Mellitus; Outcome; Patients; Peptides; Performance; Pharmacology; Physiology; Play; Positioning Attribute; Property; Protein Engineering; Proto-Oncogene Proteins c-akt; Publications; Pump; Rat-1; Rattus; Receptor Signaling; Recommendation; Research Personnel; Rest; Risk; Role; Safety; Scheme; Series; Somatostatin; Specificity; Stream; Streptozocin; Structure; Synthesis Chemistry; System; Technology; Time; Variant; Weight Gain; Western Blotting; X-Ray Crystallography; Yang; analog; base; clinical candidate; design; diabetic rat; frontier; hormone analog; image reconstruction; in silico; in vivo; innovation; interdisciplinary approach; male; mathematical model; multidisciplinary; next generation; novel; operation; particle; patient subsets; phosphoproteomics; prototype; simulation; structural biology; sugar; three dimensional structure

Project Summary Insulin and glucagon play central roles in metabolic homeostasis with long-standing application to the clinical management of diabetes mellitus (DM). This MPI application focuses on the development of glucose-responsive analogs of these hormones. The proposed technology promises to enhance the safety and efficacy of hormone replacement therapy, including in innovative bihormonal pumps in closed-loop systems. This is a key frontier of molecular pharmacology and non-standard protein engineering. The multidisciplinary MPI team encompasses protein design, biophysics, structural biology, animal physiology, clinical endocrinology, and computer simulations of mammalian metabolism. Animal studies will be performed in normal and STZ rats under the guidance of Prof. F. Ismail-Beigi (Subcontract to CWRU); computer-based interpretation of these studies as part of a design cycle will be undertaken in simulated models by Prof. M. Strano and coworkers (Subcontract to MIT). Cryo-EM studies of variant insulin-insulin receptor (IR) complexes will be performed by Prof. M.C. Lawrence (Subcontract to WEHI, Melbourne AU). The MPI team has recent joint publications, including in Nature Chemistry, J. Biol. Chem. and Diabetes. Glucose-responsive insulin (GRI) analogs are envisioned as a technology to attenuate IR signaling under conditions of hypoglycemia; glucose-responsive glucagon (GRG) analogs are envisioned as a complementary technology to attenuate glucagon-receptor (GlR) signaling under conditions of hyperglycemia. Respective protein design rests upon two complementary premises: Hypothesis 1: That development of an appropriate glucose-binding element (GBE) will enable construction of a glucose-regulated conformational switch between a glucose-free closed (inactive) state and a glucose-bound open (active) state in accord with how WT insulin binds to and activates the IR; and Hypothesis 2: That development of a distinct GBE will enable construction of a glucose-regulated conformational switch between a glucose-bound inactive state and a glucose-free active state in accordance with how WT glucagon binds to and activates the GlR. In each case the GBEs will exploit the diol-binding properties boronic acids and benzoxaboroles. Binding of glucose in a GRI activates the hormone whereas binding of glucose in a GRG inactivates the hormone. Aims 1- 3 focus on GRIs whereas Aim 4 extends our approach to GRGs. These technologies may markedly enhance the long-term health of patients with T1D and a subset of patients with T2D. Protein design will be based on classical crystal structures of insulin and glucagon, extended by dramatic recent advances in the structural biology of the IR, GlR and their respective ligand complexes. Salient structural differences between these systems promise to enable construction of opposing switches. An interdisciplinary team Approach is proposed within integrated MPI Management Plan.

项目概要 胰岛素和胰高血糖素在代谢稳态中发挥着核心作用，长期应用于临床 糖尿病（DM）的管理。该 MPI 应用重点关注葡萄糖响应性药物的开发 这些激素的类似物。所提出的技术有望提高激素的安全性和有效性 替代疗法，包括闭环系统中的创新双激素泵。这是一个关键前沿 分子药理学和非标准蛋白质工程。 多学科 MPI 团队涵盖蛋白质设计、生物物理学、结构生物学、动物生理学、 临床内分泌学和哺乳动物新陈代谢的计算机模拟。动物研究将在 F. Ismail-Beigi 教授（分包给 CWRU）指导下的正常大鼠和 STZ 大鼠；基于计算机的 M. Strano 教授将在模拟模型中对这些研究进行解释，作为设计周期的一部分 和同事（分包给麻省理工学院）。变体胰岛素-胰岛素受体 (IR) 复合物的冷冻电镜研究将 由 M.C. 教授表演Lawrence（分包给澳大利亚墨尔本 WEHI）。 MPI 团队最近联合 出版物，包括《自然化学》、《生物杂志》。化学。和糖尿病。 葡萄糖反应性胰岛素 (GRI) 类似物被设想为一种在以下情况下减弱 IR 信号传导的技术 低血糖情况；葡萄糖反应性胰高血糖素 (GRG) 类似物被认为是一种补充 在高血糖条件下减弱胰高血糖素受体（GlR）信号传导的技术。各自 蛋白质设计基于两个互补的前提： 假设 1：开发适当的葡萄糖结合元件（GBE）将使 构建无葡萄糖闭合（非活性）状态之间的葡萄糖调节构象转换 以及与 WT 胰岛素如何结合并激活 IR 一致的葡萄糖结合开放（活性）状态；和 假设 2：独特 GBE 的发展将能够构建葡萄糖调节的 葡萄糖结合非活性状态和无葡萄糖活性状态之间的构象转换 根据 WT 胰高血糖素如何结合并激活 GlR。 在每种情况下，GBE 都会利用硼酸和苯并氧杂硼杂环己烷的二醇结合特性。绑定 GRI 中的葡萄糖激活激素，而 GRG 中的葡萄糖结合则使激素失活。目标 1- 3 重点关注 GRI，而目标 4 将我们的方法扩展到 GRG。这些技术可能会显着增强 1 型糖尿病患者和一部分 2 型糖尿病患者的长期健康状况。 蛋白质设计将基于胰岛素和胰高血糖素的经典晶体结构，并通过最近的戏剧性扩展进行扩展 IR、GlR 及其各自配体复合物的结构生物学进展。结构突出 这些系统之间的差异有望构建相反的开关。跨学科 综合 MPI 管理计划中提出了团队方法。