上海交通大学学报（医学版）

›› 2012, Vol. 32 ›› Issue (5): 684-.doi: 10.3969/j.issn.1674-8115.2012.05.034

• 技术与方法 • 上一篇    

N-乙酰-β-D-氨基葡萄糖苷酶新型底物的合成及在肾小管疾病诊断中的应用

徐 蕾1, 姚莉韵2, 刘慧中3, 张建华2, 杨婉花1   

  1. 上海交通大学 1.医学院附属瑞金医院药剂科, 上海 200025； 2.医学院药学系, 上海 200025； 3.医学院化学生物学实验中心, 上海 200025
  • 出版日期:2012-05-28 发布日期:2012-06-01
  • 通讯作者: 姚莉韵, 电子信箱: yaoliyun517@yahoo.com.cn。
  • 作者简介:徐 蕾（1989—）, 女, 药师, 学士；电子信箱: 2006xulei@163.com。
  • 基金资助:

    上海市自然科学基金（11ZR1419200）

Synthesis of novel substrate for N-acetyl-β-D-glucosaminidase and its value in diagnosis of renal tubular lesions

XU Lei1, YAO Li-yun2, LIU Hui-zhong3, ZHANG Jian-hua2, YANG Wan-hua1   

  1. 1.Department of Pharmacy, Ruijin Hospital, 2.Department of Pharmacy, 3.Laboratory for Chemical Biology, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China
  • Online:2012-05-28 Published:2012-06-01
  • Supported by:

    Shanghai Natural Science Foundation, 11ZR1419200

PDF (PC)

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摘要：

目的 优化N-乙酰-β-D-氨基葡萄糖苷酶（NAG）的新型底物6-甲基-2-吡啶基-1-硫-N-乙酰-β-D-氨基葡萄糖苷（MPT-NAG）的合成方法，探讨其在肾小管疾病诊断中的应用价值。方法 采用活泼中间物（1-氯-1-脱氧乙酰糖）与6-甲基-2-巯基吡啶成苷合成底物；以MPT-NAG为底物，采用全自动生化仪测定糖尿病肾病、肾病综合征、肾移植患者和正常对照者尿液样本中NAG的活性。结果 底物MPT-NAG的结构经元素分析和1HNMR谱鉴定，高效液相色谱（HPLC）检测纯度达99.8%。NAG活性测定结果表明，糖尿病肾病、肾病综合征和肾移植患者的尿NAG活性增高，与正常对照者比较差异有统计学意义（P<0.01）。结论 经优化制备工艺合成获得NAG新型底物MPT-NAG，所用原料成本低、操作简便且产物得率较高，在肾小管疾病的诊断中具有临床应用价值。

关键词: N-乙酰-β-D-氨基葡萄糖苷酶, 6-甲基-2-吡啶基-1-硫-N-乙酰-β-D-氨基葡萄糖苷, 肾小管疾病

Abstract:

Objective To optimize the synthesis approach of 6-methyl-2-pyridyl N-acetyl-1-thio-β-D-glucosaminide (MPT-NAG) as a novel substrate for N-acetyl-β-D-glucosaminidase (NAG), and explore its value in diagnosis of renal tubular lesions. Methods The substrate was prepared by the use of 1-chloro-1-deoxy-tetraacetyl-D-glucosamine and 2-mercato-6-methylpyridine in glycoside reaction, and urinary NAG activities of samples from patients with diabetic nephropathy, patients with nephrotic syndrome, patients with renal transplantation and normal controls were detected by automated analytical device with MPT-NAG as substrate. Results The structure of the substrate of MPT-NAG was identified by elemental analysis and 1HNMR, and the purity was 99.8% determined by high performance liquid chromatography (HPLC). It was revealed that the urinary NAG activities in patients with diabetic nephropathy, nephrotic syndrome and renal transplantation were significantly higher than those in the normal group (P<0.01). Conclusion The synthesized novel substrate of MPT-NAG obtained by optimized approach with low cost and high production can be used in the diagnosis of renal tubular lesions.

Key words: N-acetyl-β-D-glucosaminidase, 6-methyl-2-pyridyl N-acetyl-1-thio-β-D-glucosaminide, renal tubular lesion