微波液相法合成4-卤代-2′,4′,5′三甲氧基查尔酮类衍生物及体外抗宫颈癌活性研究
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摘要
目的以异甘草素为先导化合物,采用微波液相法合成3种4-卤代-2′,4′,5′三甲氧基查尔酮类衍生物,研究其体外抗宫颈癌活性和促凋亡作用。方法以2,4,5-三甲氧基苯乙酮、4-氟苯甲醛、4-氯苯甲醛和4-溴苯甲醛为原料,在KOH/EtOH反应条件下,采用微波液相法合成3种4-卤代-2′,4′,5′三甲氧基查尔酮类衍生物;以人宫颈癌SiHa和HeLa细胞及中国仓鼠正常卵巢细胞CHO建立为体外模型,采用MTT法测定3种衍生物对3种细胞的增殖抑制活性,采用流式细胞仪法测定3种衍生物对SiHa细胞的促凋亡作用。结果合成得到4-氟-2′,4′,5′-三甲氧基查尔酮(化合物Ⅰ,反应时间110s,产率85.7%),4-氯-2′,4′,5′-三甲氧基查尔酮(化合物Ⅱ,反应时间90s,产率95.8%)和4-溴-2′,4′,5′-三甲氧基查尔酮(化合物Ⅲ,反应时间120s,产率90.7%)。当浓度在1、10、25、50、75、100μg/mL,作用时间在24、48、72h时,化合物Ⅰ～Ⅲ对SiHa、HeLa和CHO细胞的增殖抑制活性在0.18～97.94%,其中化合物Ⅱ对SiHa细胞的增殖抑制活性较化合物Ⅰ和Ⅲ高,但对CHO细胞的抑制活性最低。化合物Ⅱ对SiHa细胞作用24、48、72h的IC_(50)值分别是9.98、5.23和3.50μg/mL,低于异甘草素(75.48、58.29和52.21μg/mL)和顺铂(33.35、8.45和3.99μg/mL),而对CHO细胞的IC_(50)值均大于100μg/mL。化合物Ⅱ对SiHa细胞在浓度1、5、10、15、20、30μg/mL,作用24h时的凋亡率为0.3、24.8、48.7、52.9、59.9、62.5%,异甘草素和顺铂对SiHa细胞在浓度为30μg/mL,作用24h时的凋亡率为33.8和57.4%。结论采用微波液相法合成3种4-卤代-2′,4′,5′三甲氧基查尔酮类衍生物极大地缩短了反应时间、提高了产率、副产物少且安全可靠。化合物Ⅰ～Ⅲ对SiHa和HeLa细胞具有较强的增殖抑制活性,而对CHO细胞抑制活性较弱,其中化合物Ⅱ对SiHa细胞的抑制活性最强,且对SiHa细胞具有较强的促凋亡作用。
Objective Three kinds of 4-halo-2′,4′,5′trimethoxychalcone derivatives were synthesized by microwave liquid phase method with isoglycyrrhizin as lead compound to study its anti-cervical cancer activity and promote apoptosis in vitro.Methods Using 2,4,5-trimethoxyacetophenone,4-fluorobenzaldehyde,4-chlorobenzaldehyde and 4-bromobenzaldehyde as raw materials,Synthesis of three 4-halo-2′,4′,5′trimethoxychalcone derivatives by microwave liquid phase reaction under KOH/EtOH reaction conditions.Ⅰn vitro models were established for human cervical cancer HeLa and SiHa cells and Chinese hamster ovary cells CHO.The proliferation inhibitory activities of the three derivatives on the three kinds of cells were determined by MTT assay.The apoptosis-inducing effects of the three derivatives on SiHa cells were determined by flow cytometry.Results 4-Fluoro-2′,4′,5′-trimethoxychalcone(Compound Ⅰ,reaction time 110 s,yield 85.7%),4-Chloro-2′,4′,5′-trimethoxychalcone(Compound Ⅱ,reaction time 90 syield 95.8%)and4-bromo-2′,4′,5′-trimethoxychalcone(compound Ⅲ,reaction time 120 s,yield 90.7%)were synthesized by the above method.When the concentration was 1,10,25,50,75,100μg/mL and the action time was24,48,72 h,the inhibitory activities of compounds Ⅰ～Ⅲ against SiHa,HeLa and CHO cells were 0.18-97.94%.Among them,the inhibitory activity of Compound Ⅱ on SiHa cells was higher than that of Compounds Ⅰ and Ⅲ,and the inhibitory activity against CHO cells was the lowest.The IC_(50) values of Compound Ⅱ on SiHa cells at 24,48,and 72 hwere 9.98,5.23,and 3.50μg/mL,significantly lower than isoglycyrrhizin(75.48,58.29 and 52.21μg/mL)and cisplatin(33.35,8.45,3.99μg/mL),while the IC_(50) values for CHO cells were all greater than 100μg/mL.The apoptosis rates of Compound Ⅱ on SiHa cells at concentrations of 1,5,10,15,20,30μg/mL for 24 hwere 0.3,24.8,48.7,52.9,59.9,and 62.5%.The apoptotic rates of isoglycyrrhizin and cisplatin on SiHa cells at a concentration of 30μg/mL for 24hwere33.8and 57.4%,respectively.Conclusion The synthesis of three 4-halo-2′,4′,5′trimethoxychalcone derivatives by microwave liquid phase method greatly shortens the reaction time,improves the yield,and has fewer by-products and is safe and reliable.Compounds Ⅰ～Ⅲ have strong proliferation inhibitory activity on SiHa and HeLa cells,but have weaker inhibitory activity on CHO cells.Among them,compound Ⅱ has the strongest inhibitory activity on SiHa cells and has a strong pro-apoptotic effect on SiHa cells.
Objective Three kinds of 4-halo-2′,4′,5′trimethoxychalcone derivatives were synthesized by microwave liquid phase method with isoglycyrrhizin as lead compound to study its anti-cervical cancer activity and promote apoptosis in vitro.Methods Using 2,4,5-trimethoxyacetophenone,4-fluorobenzaldehyde,4-chlorobenzaldehyde and 4-bromobenzaldehyde as raw materials,Synthesis of three 4-halo-2′,4′,5′trimethoxychalcone derivatives by microwave liquid phase reaction under KOH/EtOH reaction conditions.Ⅰn vitro models were established for human cervical cancer HeLa and SiHa cells and Chinese hamster ovary cells CHO.The proliferation inhibitory activities of the three derivatives on the three kinds of cells were determined by MTT assay.The apoptosis-inducing effects of the three derivatives on SiHa cells were determined by flow cytometry.Results 4-Fluoro-2′,4′,5′-trimethoxychalcone(Compound Ⅰ,reaction time 110 s,yield 85.7%),4-Chloro-2′,4′,5′-trimethoxychalcone(Compound Ⅱ,reaction time 90 syield 95.8%)and4-bromo-2′,4′,5′-trimethoxychalcone(compound Ⅲ,reaction time 120 s,yield 90.7%)were synthesized by the above method.When the concentration was 1,10,25,50,75,100μg/mL and the action time was24,48,72 h,the inhibitory activities of compounds Ⅰ～Ⅲ against SiHa,HeLa and CHO cells were 0.18-97.94%.Among them,the inhibitory activity of Compound Ⅱ on SiHa cells was higher than that of Compounds Ⅰ and Ⅲ,and the inhibitory activity against CHO cells was the lowest.The IC_(50) values of Compound Ⅱ on SiHa cells at 24,48,and 72 hwere 9.98,5.23,and 3.50μg/mL,significantly lower than isoglycyrrhizin(75.48,58.29 and 52.21μg/mL)and cisplatin(33.35,8.45,3.99μg/mL),while the IC_(50) values for CHO cells were all greater than 100μg/mL.The apoptosis rates of Compound Ⅱ on SiHa cells at concentrations of 1,5,10,15,20,30μg/mL for 24 hwere 0.3,24.8,48.7,52.9,59.9,and 62.5%.The apoptotic rates of isoglycyrrhizin and cisplatin on SiHa cells at a concentration of 30μg/mL for 24hwere33.8and 57.4%,respectively.Conclusion The synthesis of three 4-halo-2′,4′,5′trimethoxychalcone derivatives by microwave liquid phase method greatly shortens the reaction time,improves the yield,and has fewer by-products and is safe and reliable.Compounds Ⅰ～Ⅲ have strong proliferation inhibitory activity on SiHa and HeLa cells,but have weaker inhibitory activity on CHO cells.Among them,compound Ⅱ has the strongest inhibitory activity on SiHa cells and has a strong pro-apoptotic effect on SiHa cells.
引文
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[3]FITZMAURICE C,ALLEN C,BARBER R M,et al.Global regional and national cancer incidence,mortality,years of life lost,years lived with disability,and disability-adjusted lifeyears for 29 cancer groups,1990 to 2016:a systematic analysis for the global burden of disease study[J].Jama Oncology,2018,4(11):1553-1568.
[4]TORRE L A,BRAY F,SIEGEL R L,et al.Global cancer statistics,2012[J].CA Cancer J Clin,2015,65(2):87-108.
[5]罗佳,陈金燕,曹丽芝,等.393例肿瘤患者药品不良反应报告分析[J].中国医院用药评价与分析,2014,14(11):1038-1041.
[6]谭书想.常见铂类抗肿瘤药物耐药机制和不良反应的比较分析[J].中南药学,2015,16(5):517-520.
[7]李淑青,陈亚萍.卵巢癌对顺铂和紫杉醇耐药的分子机制[J].国际妇产科学杂志,2016,43(2):145-150.
[8]YAN J,JIE C,ZHANG S,et al.Synthesis evaluation and mechanism study of novel indole chalcone derivatives exerting effective antitumor activity through microtubule destabilization in vitro and in vivo[J].J Med Chem,2016,59(11):5264-5283.
[9]GOMES M N,BRAGA R C,GRZELAK E M,et al.QSAR-driven design,synthesis and discovery of potent chalcone derivatives with antitubercular activity[J].Eur J Med Chem,2017,137(8):126-138.
[10]VIJAVAB R M,HUNG H Y,KUO P C,et al.Synthesis and biological evaluation of chalcone,dihydrochalcone,and1,3-diarylpropane analogs as anti-inflammatory agents[J].Bio Med Chem Let,2017,27(7):1547-1550.
[11]COA J C,GARCIA E,CARDA M,et al.Synthesis,leishmanicidal,trypanocidal and cytotoxic activities of quinolinechalcone and quinoline-chromone hybrids[J].Med Chem Res,2017,26(7):1-10.
[12]SHUKLA P,SATYANARAYANA M,VERMA P C,et al.Chalcone-based aryloxypropanolamine as a potential antidiabetic and antidyslipidaemic agent[J].Current Sci,2017,112(8):1675-1689.
[13]SHUKLA S,GAHLOT P,KHANDEKAR A,et al.Exploring coumarin and chalcone analogues as potential antimycobacterial agents[J].Anti Infective Agents,2018,15(2):69-86.
[14]ZHAO D H,WANG Y C,ZHENG L W,et al.Antidepressant-like effect of a chalcone compound,DHIPC and its possible mechanism[J].Iranian J Pharm Res,2018,17(1):193-201.
[15]田奇锋,姚俊.异甘草素抗肿瘤作用机制中相关信号通路的研究进展[J].海南医学,2017,28(9):1469-1471.
[16]杨旭超,西力扎提·阿不来提,木合布力·阿布力孜,等.3-甲氧基异甘草素的合成及其对宫颈癌细胞的影响[J].新疆医科大学学报,2016,39(2):173-178.
[17]COSTA A,CHIARADIA-DELATORRE L D,SANTOSBUBNIAK L,et al.Apoptotic effect of synthetic 2′,4′,5′-trimethoxychalcones in human K562 and jurkat leukemia cells[J].Med Chem Res,2014,23(10):4301-4319.
[18]BELLO M L,CHIARADIA L D,DIAS L R S,et al.Trimethoxy-chalcone derivatives inhibit growth of Leishmania braziliensis:Synthesis,biological evaluation,molecular modeling and structure-activity relationship(SAR)[J].Bio Med Chem,2011,19(16):5046-5052.