Development of enzymatic tools for rapid measurement of Advanced Glycation End Product-protein adducts

开发用于快速测量高级糖基化终产物-蛋白质加合物的酶工具

• 批准号: 10757215
• 负责人: Aaron Cravens
• 金额: $ 27.76万
• 依托单位: REVEL PHARMACEUTICALS INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10757215
• 关键词: Address; Advanced Glycosylation End Products; Aging; Amino Acids; Animal Model; Atherosclerosis; Biological Assay; Biological Markers; Biology; Blood; Blood Glucose; Characteristics; Chemicals; Clinical; Complications of Diabetes Mellitus; Coupled; Data; Detection; Development; Diabetes Mellitus; Diabetic Nephropathy; Diabetic Retinopathy; Diagnosis; Diagnostic; Digestion; Disease; Engineering; Enzymes; Excision; FDA approved; Functional disorder; Future; Glucose; Glycobiology; Glycosylated hemoglobin A; Goals; Hemoglobin; Horseradish Peroxidase; Human; Hydrogen Peroxide; Inflammatory; Kinetics; Ligation; Longevity; Lysine; Maillard Reaction; Measurement; Measures; Medicine; Methodology; Methods; Modeling; Modification; Molecular; Monitor; N(6)-carboxymethyllysine; National Institute on Aging; Organ; Oxidases; Pathogenesis; Pathologic; Patients; Peptide Hydrolases; Performance; Pharmacologic Substance; Phase; Physiological; Play; Production; Proteins; Reaction; Reagent; Request for Applications; Research; Resistance; Risk; Risk Marker; Role; Sampling; Serum; Signal Pathway; Signal Transduction; Small Business Innovation Research Grant; Sorting; Specificity; Techniques; Testing; Tissue Sample; Tissues; Variant; Vascular Diseases; adduct; age related; clinical predictors; design; detection assay; genetic selection; glycation; glyoxylate; human tissue; improved; in vitro Assay; innovation; macromolecule; macula; mortality; programs; proliferative diabetic retinopathy; prototype; rapid technique; rapid test; receptor for advanced glycation endproducts; repaired; response; risk prediction; screening; sugar; tool

PROJECT SUMMARY Advanced glycation end product (AGE) accumulation is a hallmark of mammalian aging. AGEs play causative roles in various age-related diseases by interfering with cell signaling pathways and forming adducts on cellular macromolecules, contributing to their inactivation and pathophysiology in many tissues/organs. Nε-carboxymethyl-lysine (CML), a maillard reaction product of lysine, is the best characterized AGE and is used clinically for predictive screening of diabetic kidney disease and as a biomarker for diabetic retinopathy and macular oedema. Despite the importance of CML as a biomarker of and contributor to age related disease, there are currently no techniques available for rapid, quantitative detection of CML in patient samples. Therefore, improved methods for measuring CML would be highly significant for advancing the study and treatment of diseases of aging. Revel Pharmaceuticals proposes to develop enzymes designed to cleave CML, thus enabling reagents for enzyme-based detection of one of the most important AGEs in aging, diabetes and glycobiology. Development of a convenient enzymatic method for measurement of CML in clinical samples would be an important step forward in diagnostic medicine. Revel has discovered a CML oxidase enzyme, capable of oxidizing CML to lysine. The newly discovered CML oxidase provides an opportunity to develop enzymes for diagnostic use in monitoring CML-modifications. The goal of this Phase I SBIR is to develop an enzymatic assay for rapid measurement of CML. The proposed assay will be developed analogous to the enzymatic HbA1c test which is used to assess patient blood sugar levels and diagnose diabetes. In Phase II, we will develop a workflow for CML detection in patient samples.

项目摘要 高级糖基化终产物（年龄）积累是哺乳动物衰老的标志。年龄发挥因果关系 通过干扰细胞信号通路并在细胞上形成加合物，在各种年龄相关疾病中的作用 大分子，有助于许多组织/器官的失活和病理生理学。 Nε-羧甲基赖氨酸（CML）是赖氨酸的Maillard反应产物，是最佳的年龄，IS 在临床上用于预测筛查糖尿病肾脏疾病，并作为糖尿病性视网膜病的生物标志物 和黄斑水肿。尽管CML是对年龄相关疾病的生物标志物和贡献者的重要性，但 目前尚无用于快速，定量检测患者样品中CML的技术。 因此，改进的测量CML的方法对于推进研究和 治疗衰老疾病。 Revel Pharmaceuticals的建议开发旨在清除CML的酶，从而使试剂适合 基于酶的衰老，糖尿病和糖生物学中最重要的年龄之一的检测。发展 在临床样品中测量CML的方便酶促方法将是重要的步骤 诊断医学前进。 Revel发现了一种CML氧化酶，能够将CML氧化为赖氨酸。新发现的CML 氧化酶提供了开发用于监测CML修饰的诊断使用酶的机会。这 该阶段I SBIR的目标是开发一种酶法测定，以快速测量CML。提议 测定将开发类似于用于评估患者血糖的酶促测试的测定 水平和诊断糖尿病。在第二阶段，我们将开发一个用于患者样品中CML检测的工作流程。