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: 10443890
• 负责人: FARAMARZ ISMAIL-BEIGI
• 金额: $ 49.03万
• 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 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10443890
• 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; 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.

项目摘要 胰岛素和胰高血糖素在代谢稳态中起着核心作用，并长期使用临床 糖尿病的管理（DM）。该MPI应用的重点是葡萄糖响应的发展 这些激素的类似物。提出的技术有望增强激素的安全性和功效 替代疗法，包括闭环系统中创新的生物荷尔泵。这是 分子药理学和非标准蛋白工程。 多学科MPI团队包括蛋白质设计，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代谢的计算机模拟。动物研究将在 在F. Ismail-Beigi教授的指导下（分包给CWRU）的指导下的正常和STZ大鼠；基于计算机 将这些研究解释为设计周期的一部分，将在M. Strano教授的模拟模型中进行。 和同事（分包合同）。变异胰岛素胰岛素受体（IR）复合物的冷冻EM研究将是 由M.C.教授执行劳伦斯（分包给Wehi，Melbourne au）。 MPI团队最近有联合 出版物，包括自然化学，J。Biol。化学和糖尿病。 葡萄糖响应性胰岛素（GRI）类似物被设想为在下降的技术 低血糖的条件；葡萄糖响应性胰高血糖素（GRG）类似物被设想为互补 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前提： 假设1：适当的葡萄糖结合元件（GBE）的发展将启用 在无葡萄糖封闭状态之间构建葡萄糖调节的构象切换 与WT胰岛素如何与IR结合并激活IR，并与葡萄糖结合的开放状态（活跃）状态；和 假设2：独特的GBE的发展将实现葡萄糖调节的构建 在葡萄糖结合的非活性状态与无葡萄糖活性状态之间的构象切换 根据WT胰高血糖素结合并激活GLR的方式。 在每种情况下，GBES都会利用二醇结合特性硼酸和苯唑替洛尔斯。结合 GRI中的葡萄糖激活激素，而GRG中葡萄糖的结合使激素失活。目标1- 3专注于GRIS，而AIM 4则扩展了我们对GRG的方法。这些技术可能明显增强 T1D患者和T2D患者子集的长期健康状况。 蛋白质设计将基于胰岛素和胰高血糖素的经典晶体结构。 IR，GLR及其各自配体复合物的结构生物学的进步。显着结构 这些系统之间的差异有望实现相对开关的构建。跨学科 在集成的MPI管理计划中提出了团队方法。