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

项目总结 晚期糖基化终末产物(AGE)的积累是哺乳动物衰老的标志。年龄起着因果作用 干扰细胞信号通路和形成细胞加合物在多种年龄相关疾病中的作用 大分子，在许多组织/器官中导致其失活和病理生理学。 N-ε-羧甲基-赖氨酸是赖氨酸的美拉德反应产物，是最具特征性的年龄和 临床上用于糖尿病肾病的预测筛查和糖尿病视网膜病变的生物标志物 黄斑水肿。尽管慢性粒细胞白血病作为年龄相关疾病的生物标志物和贡献者很重要， 目前还没有快速、定量检测患者样本中慢性粒细胞白血病的技术。 因此，改进测量慢性粒细胞白血病的方法对于推进这项研究和 老年病的治疗。 Revel制药公司建议开发旨在裂解CML的酶，从而使试剂能够用于 以酶为基础的检测衰老、糖尿病和糖生物学中最重要的年龄之一。发展 建立一种简便的检测临床标本中慢性粒细胞白血病的酶促方法将是重要的一步 诊断医学的前沿。 Revel发现了一种慢性粒细胞白血病氧化酶，能够将慢性粒细胞白血病氧化成赖氨酸。新发现的慢性粒细胞白血病 氧化酶为开发用于监测慢性粒细胞白血病修饰的诊断用的酶提供了机会。这个 这一阶段SBIR的目标是开发一种快速检测慢性粒细胞白血病的酶分析方法。建议数 将开发类似于用于评估患者血糖的酶促糖化血红蛋白检测方法 水平和诊断糖尿病。在第二阶段，我们将开发一个在患者样本中检测CML的工作流程。

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.