树豆叶二苯乙烯类成分的甲醚化和体外抗肿瘤活性
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摘要
为获得具有抗肿瘤活性的甲醚化先导化合物,对树豆(Cajanus cajan)叶中的二苯乙烯类成分进行甲醚化,并测定其对人肿瘤细胞的细胞毒性。将木豆素C、树豆酮酸A、Cajanotone和木豆素与碘甲烷-碳酸钾反应制备O-甲基产物,通过波谱数据分析产物结构分别鉴定为:2-异戊烯基-3,5-二甲氧基二苯乙烯(1)、树豆酮酸A甲醚(2)、5-O-methylcajanotone(3)和3-O-甲基木豆素(4),其中化合物3是新化合物。肿瘤细胞增殖抑制实验(CCK-8法)结果表明,木豆素C对乳腺癌MDA-MB-231、宫颈癌HeLa、肝癌Hep G2、结肠癌SW480及3种非小细胞肺癌细胞(A549, NCI-H460和NCI-H1299)的半数抑制浓度IC50分别为14.4、16.1、19.6、17.4和25.7～29.6μmol L~(–1);木豆素对宫颈癌和结肠癌之外的5种肿瘤细胞有弱抑制作用,IC50为44.9～78.3μmol L~(–1);而对照的3,4′,5-三甲氧基二苯乙烯(三-O-甲基白藜芦醇)对乳腺癌MDA-MB-231、宫颈癌HeLa、结肠癌SW480和肝癌Hep G2细胞有较强抑制作用(IC50为3.0～14.5μmol L~(–1))。树豆叶二苯乙烯甲醚化衍生物1～4对7种肿瘤细胞系无明显细胞毒活性。
The aim was to obtain lead compounds with anticancer activity by O-methylation of stilbene pheolic deriatives extracted from leaves of Cajanus cajan. Four derivatives, such as longistylin C, cajanonic acid A, cajanotone and cajaninstilbene acid, were treated with iodomethane and potassium carbohydrate under mild condition. The products were purified through silica gel column chromatography and their structures were identified by spectroscopic analysis as 2-isoprenyl-3,5-dimethoxy-stilbene(1), O-methyl-cajanonic acid A(2), 5-O-methyl cajanotone(3) and 3-O-methyl cajaninstilbene acid(4), respectively. Among them, compound 3 was a new compound. Effects of 1–4 and their reactants, together with resveratrol, 4′-O-methyl resveratrol and 3,4′,5-O-trimethyl resveratrol using as positive controls, on the proliferation of the cell lines of human breast cancer(MDA-MB-231), cervical cancer(HeLa), liver cancer(HepG2), colon cancer(SW480), and non-small cell lung cancer(A549, NCI-H460 and NCI-H1299) were determined by CCK-8 assay. The results showed that longistilin C distinctly inhibited the proliferation of all the seven cell lines, with the half inhibitory concentrations(IC50) to MDA-MB-231, HeLa, HepG2, and SW480 at 14.4, 16.1, 19.6, and 17.4 μmol L~(–1)(strong suppresion), with IC50 values of 25.7–29.6 μmol L~(–1) to other three cell lines of non-small cell lung cancer; cajaninstilbene acid had a weak inhibition to HepG2, A549, NCI-H460, NCI-H1299 and MDA-MB-231 cells with IC50 as 44.9– 78.3 μmol L~(–1); tri-O-methylated resveratrol showed significantly cytotoxity against MDA-MB-231, HeLa, SW480 and HepG2 cells with IC50 of 16.1-17.4 μmol L~(–1); O-methylated stilbenes 1-4 did not exhibited obvious cytotoxicity in the cell lines aboved.
The aim was to obtain lead compounds with anticancer activity by O-methylation of stilbene pheolic deriatives extracted from leaves of Cajanus cajan. Four derivatives, such as longistylin C, cajanonic acid A, cajanotone and cajaninstilbene acid, were treated with iodomethane and potassium carbohydrate under mild condition. The products were purified through silica gel column chromatography and their structures were identified by spectroscopic analysis as 2-isoprenyl-3,5-dimethoxy-stilbene(1), O-methyl-cajanonic acid A(2), 5-O-methyl cajanotone(3) and 3-O-methyl cajaninstilbene acid(4), respectively. Among them, compound 3 was a new compound. Effects of 1–4 and their reactants, together with resveratrol, 4′-O-methyl resveratrol and 3,4′,5-O-trimethyl resveratrol using as positive controls, on the proliferation of the cell lines of human breast cancer(MDA-MB-231), cervical cancer(HeLa), liver cancer(HepG2), colon cancer(SW480), and non-small cell lung cancer(A549, NCI-H460 and NCI-H1299) were determined by CCK-8 assay. The results showed that longistilin C distinctly inhibited the proliferation of all the seven cell lines, with the half inhibitory concentrations(IC50) to MDA-MB-231, HeLa, HepG2, and SW480 at 14.4, 16.1, 19.6, and 17.4 μmol L~(–1)(strong suppresion), with IC50 values of 25.7–29.6 μmol L~(–1) to other three cell lines of non-small cell lung cancer; cajaninstilbene acid had a weak inhibition to HepG2, A549, NCI-H460, NCI-H1299 and MDA-MB-231 cells with IC50 as 44.9– 78.3 μmol L~(–1); tri-O-methylated resveratrol showed significantly cytotoxity against MDA-MB-231, HeLa, SW480 and HepG2 cells with IC50 of 16.1-17.4 μmol L~(–1); O-methylated stilbenes 1-4 did not exhibited obvious cytotoxicity in the cell lines aboved.
引文
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[14] SHAN Y, HONG T, WANG Y F, et al. Synthesis and cytotoxicity of longistylin C derivatives[J]. Chin J Nat Med, 2015, 13(4):311–315.doi:10.1016/S1875-5364(15)30021-2.
[15] WANGYN,GEP,HUYJ,etal.SynthesismethodofdiethylstilbestrolcompoundmethylpigeonpeaketonicacidA:China,CN201410032181. 6[P]. 2015-05-27.王亚农,戈平,胡英杰,等.一种菧类化合物甲基树豆酮酸A的合成方法:中国, CN201410032181.6[P]. 2015-05-27.
[16] ZHANG N L, ZHU Y H, HUANG M R, et al. Two new stilbenoids fromCajanuscajan[J].ZNaturforschB,2012, 67(12):1314–1318.doi:10.5560/znb.2012-0184.
[17] LIZR,JIXY,XUEST,etal.Cajanincompoundwithsimilar structures,aswellaspreparationmethodandapplicationthereof:China, CN201110439374.X[P]. 2013-06-26.李卓荣,季兴跃,薛司徒,等.一组木豆素结构类似化合物、制备方法和应用:中国, CN201110439374.X[P]. 2013-06-26.
[18]GOSSLAUA,CHENM,HOCT,etal.Amethoxyderivativeof resveratrol analogue selectively induced activation of the mitochondrial apoptoticpathwayintransformedfibroblasts[J].BrJCancer,2005,92(3):513–521. doi:10.1038/sj.bjc.6602300.
[2]GOTOH,YANAGIMACHIM,GOTOS,etal.Methylatedchrysin reduced cell proliferation, but antagonizedcytotoxicity of other anticancer drugs in acute lymphoblastic leukemia[J]. Anti-Cancer Drugs,2012, 23(4):417–425. doi:10.1097/CAD.0b013e32834fb731.
[3]WENGJC,YANGYT,HOTH,etal.Mechanismsofapoptotic effects induced by resveratrol, dibenzoylmethane, and their analogues on human lung carcinoma cells[J]. J Agric Food Chem, 2009, 57(12):5235–5243. doi:10.1021/jf900531m.
[4]WENG J C, WU C F, HUANG H W, et al. Evaluation of anti-invasion effect of resveratrol and related methoxy analogues on human hepatocarcinoma cells[J]. J Agric Food Chem, 2010, 58(5):2886–2894. doi:10.1021/jf904182y.
[5]HSIEH C T, WONG C, BENNETT J D, et al. Regulation of p53 and cell proliferation by resveratrol and its derivatives in breast cancer cells:An in silico and biochemical approach targeting integrinαvβ3[J]. Int J Cancer, 2011, 129(11):2732–2743. doi:10.1002/ijc.25930.
[6]FU Y J, YU P. Method for synthesizing anti-form stilbene compounds and application of method in preparing anti-tumor medicines:China,CN201010592686.X[P]. 2012-07-11.付玉杰,余平.一种合成反式芪类化合物的方法及其在制备抗肿瘤药物中的应用:中国, CN201010592686.X[P]. 2012-07-11.
[7]FUYJ,KADIOGLUO,WIENCHB,etal.Cellcyclearrestand induction of apoptosis by cajanin stilbene acid from Cajanus cajan in breast cancer cells[J]. Phytomedicine, 2015, 22(4):462–468. doi:10.1016/j.phymed.2015.02.005.
[8]CAIJZ,TANGR,YEGF,etal.Ahalogen-containingstilbene derivativefromtheleavesofCajanuscajanthatinducesosteogenic differentiation of human mesenchymal stem cells[J]. Molecules, 2015,20(6):10839–10847. doi:10.3390/molecules200610839.
[9]ASHIDI J S, HOUGHTON P J, HYLANDS P J, et al. Ethnobotanical survey and cytotoxicity testing of plants of south-western Nigeria used to treat cancer, with isolation of cytotoxic constituents from Cajanus cajan Mill. leaves[J]. J Ethnopharmacol, 2010, 128(2):501–512. doi:10.1016/j.jep.2010.01.009.
[10] CAIJZ.ChemicalconstituentsfromleavesofArdisiacrenataand Cajanuscajanandanticancereffects[D].Guangzhou:Guangzhou University of Chinese Medicine, 2012:83–100.蔡佳仲.朱砂根和树豆叶的化学成分及抗肿瘤作用研究[D].广州:广州中医药大学, 2012:83–100.
[11] SHEN X L, WANG L, HU Y J, et al. Application of pigeon pea ketonic acid A in terms of preparation of medicines for accompanied diseases of diabetes mellitus and hyperlipidaemia:China, CN201210119665.5[P]. 2012-09-19.沈小玲,王璐,胡英杰,等.树豆酮酸A在制备糖尿病伴随症及高脂血症药物中的应用:中国, CN201210119665.5[P]. 2012-09-19.
[12] GUORX.Studiesonstructuralmodificationofquercetin,isoalantolactone and alantolactone[D]. Shijiazhuang:Hebei Medical University,2014:15–17.郭瑞霞.槲皮素、异土木香内酯和土木香内酯的结构修饰研究[D].石家庄:河北医科大学, 2014:15–17.
[13] HU Y L, SHEN X L, CHEN M G, et al. Structural identification on the saponification products of raw cajanolactone A[J]. J Trop Subtrop Bot,2017, 25(1):93–97. doi:10.11926/jtsb.3617.胡阳亮,沈小玲,陈梅果,等.树豆内酯A粗品皂化产物的化学结构鉴定[J].热带亚热带植物学报,2017,25(1):93–97.doi:10.11926/jtsb.3617.
[14] SHAN Y, HONG T, WANG Y F, et al. Synthesis and cytotoxicity of longistylin C derivatives[J]. Chin J Nat Med, 2015, 13(4):311–315.doi:10.1016/S1875-5364(15)30021-2.
[15] WANGYN,GEP,HUYJ,etal.SynthesismethodofdiethylstilbestrolcompoundmethylpigeonpeaketonicacidA:China,CN201410032181. 6[P]. 2015-05-27.王亚农,戈平,胡英杰,等.一种菧类化合物甲基树豆酮酸A的合成方法:中国, CN201410032181.6[P]. 2015-05-27.
[16] ZHANG N L, ZHU Y H, HUANG M R, et al. Two new stilbenoids fromCajanuscajan[J].ZNaturforschB,2012, 67(12):1314–1318.doi:10.5560/znb.2012-0184.
[17] LIZR,JIXY,XUEST,etal.Cajanincompoundwithsimilar structures,aswellaspreparationmethodandapplicationthereof:China, CN201110439374.X[P]. 2013-06-26.李卓荣,季兴跃,薛司徒,等.一组木豆素结构类似化合物、制备方法和应用:中国, CN201110439374.X[P]. 2013-06-26.
[18]GOSSLAUA,CHENM,HOCT,etal.Amethoxyderivativeof resveratrol analogue selectively induced activation of the mitochondrial apoptoticpathwayintransformedfibroblasts[J].BrJCancer,2005,92(3):513–521. doi:10.1038/sj.bjc.6602300.