【作者】 王兵；

【导师】 王晓静；

【作者基本信息】 济南大学 ， 药学（药物化学）， 2018， 硕士

【摘要】 该论文主要从3-芳基香豆素类化合物的合成方法改进与抗糖尿病活性筛选两个大方面进行研究。通过使用微波辐射加热法合成了多种取代的3-芳基香豆素类衍生物。该方法具有环境友好、经济、分离和纯化过程简单、副产物少和较高的反应产率等特点。在合成3-芳基香豆素类化合物骨架这一部分探索了反应的最佳条件,结论为:反应功率100 W,反应时间70分钟,此时反应产率大约在96%以上。我们对44个3-芳基香豆素类化合物进行了分子对接研究和体外活性初步筛选,包括DPPH和羟基自由基清除活性、α-葡萄糖苷酶抑制活性、晚期糖基化终产物形成抑制活性。分子对接结果显示大部分3-芳基香豆素类化合物几乎完全嵌入位点口袋中,其构象很好的与结合位点口袋契合,达到结构最佳匹配。后来的体外α-葡萄糖苷酶抑制实验结果也验证了计算机辅助药物设计的对接结果,说明采用计算机辅助药物设计软件对活性初步筛选具有一定的指导意义,而且可以用最低的成本完成对实际效果进行预估,节省了实验资源。构效关系研究表明,大多数化合物表现出显著的自由基清除,并且具有邻二酚羟基结构的3-芳基香豆素类化合物更为优越。另外,母核中的7-羟基对抑制α-葡萄糖苷酶是必要的,而具有5,7-二羟基的3-芳基香豆素的α-葡糖苷酶抑制活性比具有7-羟基的3-芳基香豆素强。同时发现母核3位苯基上的4’-羟基是α-葡糖苷酶抑制活性中的另一个重要的活性基团。多数化合物对糖基化终产物(AGEs)的形成呈现较强的抑制活性,甚至优于氨基胍盐酸盐,其中3-芳基香豆素类化合物均具有4’-羟基时,那些具有7-羟基的化合物优于具有5,7-二羟基的化合物而且也比具有6-羟基的化合物更强。证明3-芳基香豆素类化合物对糖尿病及其并发症存在多个潜在的作用靶点,很可能在糖尿病及其并发症形成的多个阶段产生疗效,这对开发预防和治疗糖尿病及其并发症的多靶点药物具有指导意义。综合考虑各个化合物在多方面的活性,选取化合物8、14、22、25和32这五个化合物进行小鼠急性毒性和多次给药的降糖活性测定。首先口服毒性实验表明这三个化合物的半数致死率均大于5000 mg/kg,它们对小鼠无害。然后通过测定单次给药化合物8、14、22、25和32对正常小鼠和糖尿病小鼠血糖的影响,筛选效果较好的化合物及最适合的用药剂量。结果表明,目标化合物8和14的效果与阳性对照药品格列本脲的效果相当,而化合物32稍强于格列本脲。随后选择化合物8、14和32进行小鼠多次给药的降糖活性测定,用药剂量为30 mg/kg。经过16天的探索,从糖尿病小鼠血糖降低水平、体重变化水平与糖耐量调节水平三个方面反映其降糖活性。结果表明,这三个化合物可以明显降低糖尿病小鼠血糖水平,减轻小鼠消瘦状况,同时可以改善糖耐量调节水平,它们的效果与阳性对照药品格列本脲的差异没有统计学意义,其中化合物32降糖活性最为显著。根据以上实验,目标化合物32可作为抗糖尿病新药研发的先导化合物,用于开发多靶点抗糖尿病及其并发症的潜在药物。综上所述,本研究找到了多个具有糖尿病治疗相关活性的化合物,具有多靶点、高活性的优点,初步证明了此类化合物在糖尿病治疗方面的开发潜力,说明抗糖尿病活性在具有3-芳基香豆素骨架的天然化合物中普遍存在。更多还原

【Abstract】 This paper studies 3-arylcoumarin derivatives including its synthesis and screening of antidiabetic activity.A variety of substituted 3-arylcoumarin derivatives were synthesized by using microwave radiation heating.The method has the characteristics of environmental friendliness,economy,simple separation and purification process,less by-products and higher reaction yield.In the synthesis of the 3-arylcoumarin skeleton,the best conditions for the reaction were explored.The reaction power was 100 W and the reaction time was 70 minutes.At this point the reaction yield is approximately above 96%.Those 3-arylcoumarin derivatives were screened for antioxidant,α-glucosidase inhibitory,advanced glycation end-products（AGEs）formation inhibitory and molecular docking study.The results of molecular docking showed that most of the 3-arylcoumarin compounds were almost completely embedded in the pocket of the site,and their conformations were well matched to the binding site pocket to achieve the best structural match.The results of in vitroα-glucosidase inhibition experiment also verified the results of molecular docking,indicating that the use of computer-aided drug design software has certain guidance for the preliminary screening of the activity,and can be used to predict the actual effect with the lowest cost.So,computer-aided drug design software has the advantages of cost saving,convenience,and high efficiency.Most compounds exhibited significant antioxidant and advanced glycation end-products formation inhibitory activities.Among them,3-arylcoumarin compounds with o-diphenol hydroxy structures are more advantageous.The3-arylcoumarin with 7-hydroxyl group showed good inhibitory activity,indicating that7-hydroxyl group was necessary for inhibitingα-Glucosidase,and also is effective for free radical scavenging activity and inhibition of glycosylated end products.3-Arylcoumarin with7-hydroxyl group was weaker than that with 5,7-dihydroxy groups in meta-position.The result suggested that the 4’-hydroxyl group was another active site inα-glucosidase inhibitory activity.When the 3-arylcoumarin both have the 4’-hydroxy groups,those compounds with7-hydroxy group were better than those with 5,7-dihydroxy groups and which were stronger than those with 6-hydroxy group.The 4’-fluoro group had substantially no effect on the activity compared to the 4’-hydroxy group,but 4’-bromo group decreased the inhibitory activity.The comparison of the IC₅₀ values showed that 3-arylcoumarin with 4’-hydroxy is stronger than that with 4’-methoxy and 4’-methyl in AGEs formation inhibitory activity.On the basis of this,the compounds 8,14,22,25 and 32 were selected to carry out acute toxicity and antidiabetic activity in mice.First of all,oral toxicity experiments showed that the lethal dose of 50%of these five compounds was greater than 5000 mg/kg,so they were not deleterious to mice.And then through the acute hypoglycemic assay of normal mice and acute antihyperglycemic assay of diabetic mice to choose compounds and the most appropriate dosage.The results showed that the effects of the target compounds 8 and 14 were comparable to those of the positive control drug glibenclamide,while compound 32 was slightly stronger than glibenclamide.Thereafter,the target compounds 8,14,32（30 mg/kg）and glibenclamide were administered once on a daily,for a period of 16 days,to STZ-induced diabetic mice.The results showed that these three compounds could significantly reduce the blood glucose level of diabetic mice,reduce the weight loss of mice,and improve the level of impaired glucose tolerance.Antidiabetic activity studies had shown that compounds 8 and 14were found to be equipotent to the standard drug glibenclamide in vivo.In summary,the target compound 32 provided a potential drug design concept for the development of multi-target antidiabetic and its complications of potential drugs.The whole experiment showed that antidiabetic activity is prevalent in 3-arylcoumarins,which added a new natural skeleton to the development of antidiabetic active drugs.更多还原