新型京尼平衍生物的设计、合成及抗抑郁活性研究

Design,synthesis and antidepressant activity of novel genipin derivatives

抑郁症发病机制复杂,现有部分单胺类抗抑郁药物存在耐药或失效脱靶等问题。中药经方具有多成分、多靶点等特点,在临床中用于治疗抑郁症效果显著。越鞠丸在抑郁症治疗中成效显著,其中“栀子-川芎”药对发挥了关键的抗抑郁作用,京尼平苷和川芎嗪分别作为栀子和川芎的有效成分具有良好抗抑郁活性。本研究基于京尼平的神经保护活性及川芎嗪的快速抗抑郁活性,通过药效团拼合原理将二者进行拼合,设计合成一系列新型京尼平衍生物,并进行神经保护活性和抗抑郁作用研究。结果表明,新型京尼平衍生物在谷氨酸诱导的HT-22细胞模型上具有良好的神经保护活性,其中化合物W-1和W-3的保护活性较好。在行为绝望抑郁(behavioral despair depression,BDD)模型小鼠的悬尾实验与强迫游泳实验的研究中发现,化合物W-3相较于W-1抗抑郁活性更优。进一步对慢性不可预知性温和应激(chronic unpredictable mild stress, CUMS)模型小鼠的行为学进行研究,结果显示, W-3可显著改善模型小鼠抑郁样行为,所有动物实验经安徽中医药大学实验动物伦理委员会批准(批准号:AHUCM-mouse-2022027)。通过蛋白质印迹法分析了优选化合物W-3对蛋白激酶A (protein kinase A, PKA)、cAMP反应元件结合蛋白(cAMP response element binding, CREB)、脑源性神经营养因子(brain-derived neurotrophic factor, BDNF)、5-羟色胺1A (5-hydroxytryptamine 1A, 5-HT1A)受体以及N-甲基-D-天冬氨酸离子型谷氨酸受体2A (N-methyl-D-aspartate ionicglutamatereceptor2A,GluN2A)和N-甲基-D-天冬氨酸离子型谷氨酸受体2B(N-methyl-D-aspartateionic glutamate receptor 2B, GluN2B)蛋白表达的影响,检测结果显示, W-3可显著上调PKA、CREB、BDNF和5-HT1A蛋白表达水平,并下调GluN2A和GluN2B蛋白表达水平;qRT-PCR结果与蛋白质印迹法检测结果一致。根据以上结果推测,化合物W-3具有潜在的抗抑郁作用,作用机制可能与调节GluN2A、GluN2B及5-HT1A受体蛋白的表达,从而激活PKA-CREB-BDNF信号通路有关。

The pathogenesis of depression is complex,and some existing monoamine antidepressants have problems such as drug resistance or off-target failure.Traditional Chinese medicine has the characteristics of"multi-component and multi-target",and has been used in the treatment of depression in clinical practice.Yueju pill is effective in the treatment of depression.Geniposide and ligustrazine,the active ingredients of Gardeniae fructus and Ligusticum sinense'Chuanxiong',play a key role in the treatment of depression.In this study,based on the neuroprotective activity of genipin and the rapid antidepressant activity of tetramethylpyrazine,a series of novel genipin derivatives were designed and synthesized through pharmacophore assembly principle,and their neuroprotective activity and antidepressant effect were investigated.The results showed that the novel genipin derivatives had well neuroprotective activity on the glutamate-induced HT-22 cell model,with compounds W-1 and W-3 showing better protective activity.In behavioral despair depression(BDD)model mice,compound W-3 was found to have better antidepressant activity than W-1 in tail suspension test and forced swimming test.Further study on the behavior of chronic unpredictable mild stress(CUMS)model mice showed that W-3 could significantly improve the depression-like behavior of model mice.All animal experiments were approved by the Experimental Animal Ethics Committee of Anhui University of Chinese Medicine(approval number:AHUCMmouse-2022027).The effects of the preferred compound W-3 on protein kinase A(PKA),cAMP response element binding protein(CREB),brain-derived neurotrophic factor(BDNF),5-hydroxytryptamine 1A(5-HT,^)receptor,Nmethyl-D-aspartate ionic glutamate receptor 2A(GluN2A)and N-methyl-D-aspartate ionic glutamate receptor 2B(GluN2B)were analyzed by Western blot.W-3 treatment significantly up-regulated the protein expression of PKA,CREB,BDNF and 5-HTra,and down-regulated the protein expression of GluN2A and GluN2B.The results of qRTPCR were consistent with those of Western blot.According to the above results,compound W-3 has a potential antidepressant effect,and its mechanism may be related to the activation of PKA-CREB-BDNF signaling pathway by regulating the expression of GluN2A,GluN2B and 5-HTiA receptor proteins.