目的 建立以γ-氨基丁酸转氨酶(GABA-T)抑制剂类抗癫痫神经系统药物筛选模型,并运用此模型对天麻有效成分及其类似物进行体外活性筛选和构效关系分析。方法 优化酶催化反应温度、反应时间、底物NAD⁺浓度、底物α-酮戊二酸浓度、底物γ-氨基丁酸(GABA)浓度等因素建立GABA-T酶系活性筛选模型,采用对羟基苯甲醛(HBA)及其11种结构类似物验证模型可靠性。结果 成功构建GABA-T酶系抗癫痫活性筛选模型,HBA及11个结构类似物测定结果与文献报道抗癫痫作用相一致,且构效分析发现-OH及苯环对位上的-CHO为必需药效团。结论 建立的模型可应用于GABA-T酶抑制剂的高通量筛选,为GABA-T酶抑制剂类抗癫痫活性成分的筛选及其作用机制研究提供参考。

Objective To establish a screening model in vitro with γ-aminobutyric acid transaminase (GABA-T) as a target for screening the inhibitors of anti-epilepsy drugs for the treatment of nervous diseases and finish the structure-activity relationship analysis of p-hydroxybenzaldehyde as effective components in Gastrodiae Rhizoma and its analogs. Methods Catalyst performed temperature, reaction time, the concentration of NAD⁺, α-ketoglutarate, and GABA were investigated to optimize the model. The test of p-hydroxybenzaldehyde (HBA) and its 11 analogs were selected as verification of this model. Results The screening model in vitro based on GABA-T was established. The testing result of p-HBA and its 11 analogs was consistent with the practical activity recorded in the literature. Hydroxyl groups of -OH and -CHO in p-aldehyde on the benzene ring were pharmacophores. Conclusion The model established in this paper can be used for high throughput screening the inhibitors of anti-epilepsy drugs, which provides the reference for the study on the inhibitors of anti-epilepsy drugs for nervous diseases and their mechanisam.

国家自然科学基金资助项目(81173519);北京中医药大学中青年教师自主科研课题面上项目(2015-JYB-JSMS023);北京中医药大学科研创新团队项目(中药先导化合物发现与开发,2011-CXTD-15)