| 引用本文: | 董佩杰,张新国,刘英娟,唐鹏,苏媛,苏岷山,韩锡萍,邵男.腺苷脱氨酶抑制剂快速筛选模型的建立[J].中国现代应用药学,2015,32(11):1301-1305. |
| DONG Peijie,ZHANG Xinguo,LIU Yingjuan,TANG Peng,SU Yuan,SU Minshan,HAN Xiping,SHAO Nan.Rapid Drug Model for Screening Adenosine Deaminase Inhibitor[J].Chin J Mod Appl Pharm(中国现代应用药学),2015,32(11):1301-1305. |
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| 摘要: |
| 目的 建立腺苷脱氨酶(adenosine deaminase，ADA)抑制剂的快速筛选模型。方法 采用ADA酶，以腺苷为底物，2'-脱氧助间型霉素作为抑制剂验证反应体系，通过紫外分光光度法检测，优化反应条件，并对248种中药提取物及其内生菌发酵产物进行筛选。结果 建立了最佳模型方案为：pH 7.5的PBS缓冲液体系，底物浓度为腺苷200 μg·mL-1，ADA酶浓度55 IU·mL-1，连续检测腺苷在265 nm下OD值动态变化情况。此模型可以用于ADA酶抑制剂的快速筛选，且基于此模型共筛选获得23个有一定活性的粗提物。结论 本实验建立的体系可以为ADA酶抑制剂的筛选提供简单易行、稳定、可靠的研究平台。 |
| 关键词: 腺苷脱氨酶 抑制剂 筛选模型 |
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| 基金项目:国家自然科学基金(31360379)；国家级大学生创新创业训练计划支持(091073117) |
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| Rapid Drug Model for Screening Adenosine Deaminase Inhibitor |
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DONG Peijie1, ZHANG Xinguo1, LIU Yingjuan1, TANG Peng1, SU Yuan2, SU Minshan1, HAN Xiping1, SHAO Nan1
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1.School of Life Science and Engineering, Lanzhou University of Technology, Key Laboratory of Screening and Deep Processing in New Tibetan Medicine of Gansu Province, Lanzhou 730050, China;2.Xining Institute for Drug Control, Qinghai 810000, China
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| Abstract: |
| OBJECTIVE To establish a rapid screening model for adenosine deaminase(ADA) inhibitor. METHODS Using the bovine spleen ADA as enzyme, adenosine as substrate, pentostatin as an inhibitor of the reaction system verification, an ultraviolet spectrophotometry method was used to screen for adenosine deaminase inhibitor after optimize its reaction conditions. RESULTS The best model scheme for pH was 7.5, phosphate buffer solution(PBS) was buffer system, the concentration of the substrate concentration of adenosine was at 200 μg·mL-1, and the concentration of ADA enzymes was at 55 IU·mL-1. The model could be used for rapid screening of ADA enzyme inhibitors and 23 activities were obtained from 248 kinds of Chinese herb extracts and fermentation products endophytes screening by the present model. CONCLUSION The present method is simple, stable and reliable for screening ADA inhibitor. |
| Key words: adenosine deaminase inhibitor screening model |
