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.

项目概要 晚期糖基化终末产物 (AGE) 积累是哺乳动物衰老的一个标志。 通过干扰细胞信号传导途径并在细胞上形成加合物，在各种与年龄相关的疾病中发挥作用 大分子，有助于其在许多组织/器官中的失活和病理生理学。 Nε-羧甲基-赖氨酸 (CML) 是赖氨酸的美拉德反应产物，是表征最好的 AGE， 临床上用于糖尿病肾病的预测筛查以及作为糖尿病视网膜病变的生物标志物 尽管 CML 作为年龄相关疾病的生物标志物和促成因素很重要， 目前尚无可用于快速、定量检测患者样本中 CML 的技术。 因此，改进测量 CML 的方法对于推进研究和 治疗衰老疾病。 Revel Pharmaceuticals 提议开发旨在裂解 CML 的酶，从而使试剂能够用于 对衰老、糖尿病和糖生物学发展中最重要的 AGE 之一进行酶检测。 建立一种方便的酶法来测量临床样本中的 CML 将是重要的一步 诊断医学的进步。 Revel 发现了一种 CML 氧化酶，能够将 CML 氧化为赖氨酸。 氧化酶提供了开发用于监测 CML 修饰的诊断用途的酶的机会。 该 I 期 SBIR 的目标是开发一种用于快速测量 CML 的酶法。 将开发类似于用于评估患者血糖的酶促 HbA1c 测试的检测方法 在第二阶段，我们将开发一个用于患者样本中 CML 检测的工作流程。