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.

项目摘要 胰岛素和胰高血糖素在代谢稳态中起着重要作用，长期应用于临床 糖尿病（DM）的管理。该MPI应用程序侧重于葡萄糖响应型 这些激素的类似物。这项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。这是一个关键的前沿领域， 分子药理学和非标准蛋白质工程。 多学科的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新陈代谢的计算机模拟。动物研究将在 在F. Ismail-Beigi（分包给CWRU）;计算机化 作为设计周期的一部分，这些研究的解释将由M.斯特拉诺 和同事（分包给MIT）。将对变异胰岛素-胰岛素受体（IR）复合物进行冷冻电镜研究。 由M.C.教授执行。Lawrence（分包给WEHI，墨尔本Au）。MPI团队最近联合 出版物，包括Nature Chemistry，J. Biol. Chem. and Diabetes。 葡萄糖响应性胰岛素（GRI）类似物被设想为在胰岛素抵抗下减弱IR信号传导的技术。 低血糖症的条件;葡萄糖响应性胰高血糖素（GRG）类似物被设想为补充 在高血糖条件下减弱胰高血糖素受体（GlR）信号传导的技术。相应 蛋白质设计基于两个互补的前提： 假设1：适当的葡萄糖结合元件（GBE）的开发将使 在无葡萄糖闭合（失活）状态和无葡萄糖闭合（失活）状态之间构建葡萄糖调节的构象转换 和与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：不同GBE的发展将能够构建葡萄糖调节的 葡萄糖结合的非活性状态和无葡萄糖的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将利用硼酸和苯并氧杂硼杂环戊烯的二醇结合特性。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侧重于GRI，而Aim 4将我们的方法扩展到GRG。这些技术可以显著提高 T1 D患者和T2 D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，最近戏剧性的扩展 IR、GlR及其相应配体复合物的结构生物学进展。突出构造 这些系统之间的差异保证能够构造相对的开关。一个跨学科 在综合MPI管理计划中提出了团队方法。