| 种蕾,张亚西,王楠,等.毛细管电泳用于神经氨酸酶抑制剂筛选方法研究D[J].中国海洋药物,2014,33(6):47-51. |
| 毛细管电泳用于神经氨酸酶抑制剂筛选方法研究D |
| The method study of capillary electrophoresis used in neuraminidase inhibitors screening |
| 投稿时间：2014-06-05 修订日期：2014-07-09 |
| DOI： |
| 中文关键词: 神经氨酸酶 毛细管 可口革囊星虫 |
| English Keywords:neuraminidase capillary electrophoresis Phascolosoma esculenta |
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| 中文摘要: |
| 目的 建立适用于复杂体系的神经氨酸酶抑制剂毛细管电泳筛选方法。方法 以神经氨酸酶为靶点,以2￠-(4-甲基伞形酮-a-N-乙酰神经氨酸)为底物,建立神经氨酸酶抑制剂的毛细管电泳筛选方法,并将该方法用于海洋药物可口革囊星虫粗提物的活性筛选。结果 以10 mmol.L-1 pH 10.0的硼砂为背景电解质,酶促反应底物与产物在4.5分钟内基线分离。酶促反应在含4 mmol.L-1 CaCl2的pH 3.5的磷酸体系中,37℃水浴中酶与底物孵育15min,产物有稳定的较大的生成量。扎那米韦阳性对照验证结果表明,该方法可用于神经氨酸酶抑制剂筛选。对源于可口革囊星虫的11个粗提物筛选结果显示,7个粗提物对神经氨酸酶有抑制活性。结论 系统优化了毛细管电泳分离分析方法及神经氨酸酶反应体系,建立的模型可用于复杂体系中抗流感病毒药物神经氨酸酶抑制剂的筛选。 |
| English Summary: |
| Abstract: Objective To establish the capillary electrophoresis methods for screening neuraminidase inhibitors. Methods A method for neuraminidase inhibitor screening by capillary electrophoresis is described. The neuraminidase as target and 2?-(4-methyl umbelliferone-?-N-acetylneuraminic acid) as substrate. Finally, screening 19 natural extracts of Phascolosoma esculenta was demonstrated. Result With borax solution whose concentrtion is 10mmol?L-1 and pH is 10.0 as background electrolyte, the substrate and product of enzymatic reaction were baseline separated within 4.5 minutes. The product is stable and generated largely when the enzymatic reaction reacts in the phosphoric acid system with 4 mmol?L-1 CaCl2 at pH 3.5, and the enzyme and substrate were incubated for 15 min in the 37℃ water bath. The zanamivir positive control verification results show that the method can be used for neuraminidase inhibitor screening. The screening results of 11 crude extracts from Phascolosoma esculenta show that 7 components have inhibitory activity on neuraminidase. Conclusion Systematcally optimized the capillary electrophoresis analysis method and the neuraminidase reaction system, the model established can be used for screening neuraminidase inhibitors of anti influenza virus drugs from complex systems. |
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