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Cytotoxicity of benzophenanthridine alkaloids from the roots of Zanthoxylum nitidum (Roxb.) DC. var. fastuosum How ex Huang ab
a
a
b
Cheng-Fang Wang , Li Fan , Mei Tian , Shu-Shan Du , Zhi-Wei c
a
b
Deng , Jiang-Bin Feng , Yong-Yan Wang & Xu Su
a
a
Click for updates
China CDC Key Laboratory of Radiological Protection and Nuclear Emergency, National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention, Xicheng District, Beijing 100088, P.R. China b
State Key Laboratory of Earth Surface Processes and Resource Ecology, Beijing Normal University, No.19 Xinjiekouwai Street, Haidian District, Beijing 100875, P.R. China c
Analytic and Testing Center, Beijing Normal University, No.19 Xinjiekouwai Street, Haidian District, Beijing 100875, P.R. China Published online: 03 Feb 2015.
To cite this article: Cheng-Fang Wang, Li Fan, Mei Tian, Shu-Shan Du, Zhi-Wei Deng, Jiang-Bin Feng, Yong-Yan Wang & Xu Su (2015) Cytotoxicity of benzophenanthridine alkaloids from the roots of Zanthoxylum nitidum (Roxb.) DC. var. fastuosum How ex Huang, Natural Product Research: Formerly Natural Product Letters, 29:14, 1380-1383, DOI: 10.1080/14786419.2014.1002090 To link to this article: http://dx.doi.org/10.1080/14786419.2014.1002090
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or
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This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions
Natural Product Research, 2015 Vol. 29, No. 14, 1380–1383, http://dx.doi.org/10.1080/14786419.2014.1002090
SHORT COMMUNICATION Cytotoxicity of benzophenanthridine alkaloids from the roots of Zanthoxylum nitidum (Roxb.) DC. var. fastuosum How ex Huang Cheng-Fang Wangab, Li Fana, Mei Tiana, Shu-Shan Dub, Zhi-Wei Dengc, Jiang-Bin Fenga, Yong-Yan Wangb and Xu Sua*
Downloaded by [New York University] at 11:38 19 June 2015
a
China CDC Key Laboratory of Radiological Protection and Nuclear Emergency, National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention, Xicheng District, Beijing 100088, P.R. China; bState Key Laboratory of Earth Surface Processes and Resource Ecology, Beijing Normal University, No.19 Xinjiekouwai Street, Haidian District, Beijing 100875, P.R. China; cAnalytic and Testing Center, Beijing Normal University, No.19 Xinjiekouwai Street, Haidian District, Beijing 100875, P.R. China (Received 20 October 2014; final version received 18 December 2014)
This work aimed to investigate benzophenanthridine from the roots of Zanthoxylum nitidum (Roxb.) DC. var. fastuosum How ex Huang for the first time. Thirteen benzophenanthridines were isolated, and our results of the cytotoxic activities indicated that compound 6 exhibited the best potency against A549, Hela, SMMC-7721 and EJ, with the IC50 values of 27.50, 37.50, 16.95 and 60.42 mM, respectively. Compounds 7 and 11 also showed strong cytotoxicity when tested against the four human cancer cell lines (A549, Hela, SMMC-7721 and EJ), while only compounds 12 and 13 displayed cytotoxicity in inhibiting BALL-1 proliferation among all the compounds. These results suggested that benzophenanthridines may become a valid alternative of potential basis for new anti-proliferative agents. Keywords: benzophenanthridine alkaloids; Zanthoxylum nitidum (Roxb.) DC. var. fastuosum How ex Huang; cytotoxic activity
1. Introduction The genus Zanthoxylum is very well known for its diversified chemistry, particularly by the presence of alkaloids, aromatic and aliphatic amides, and phenylpropanoids – lignans and coumarins (Mester et al. 1983; Simanek & Brossi 1985). Among them, benzophenanthridine alkaloids are particularly abundant in roots and stem bark (Krane et al. 1984; Ng et al. 1987; Hu et al. 2006; Luo et al. 2012), which have shown broad spectrum of biological activities, including cytotoxic, antiviral, antibacterial, antinociceptive, anti-inflammatory and insecticidal properties (Vanquelef et al. 2004; Hu et al. 2006; Ueno 2008; Obiang-Obounou et al. 2011; Mansoor et al.
*Corresponding author. Email: [email protected] q 2015 Taylor & Francis
Natural Product Research
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Downloaded by [New York University] at 11:38 19 June 2015
2013). Zanthoxylum nitidum (Roxb.) DC. var. fastuosum How ex Huang (Z. nitidum var. fastuosum) is widely used to substitute Z. nitidum (Roxb.) DC. in folk medicine for the treatment of ulcer, toothache, stomachache and traumatic injuries, which was found in Guangxi province of China primarily (Wang et al. 2006). However, very few information on its chemical constituents and biological activities was reported in previous study (Wang et al. 2006; Hu et al. 2008; Sun et al. 2011). Further investigation is necessary to find the chemical basis of activities in this plant. In this work, we focus on benzophenanthridine alkaloids isolated from the roots of Z. nitidum var. fastuosum, aiming to identify the active compounds by assessing their cytotoxic activities against BALL-1, A549, Hela, SMMC-7721 and EJ cell lines in vitro. The correlation between structure and cytotoxicity is also discussed.
2. Results and discussion Thirteen benzophenanthridines were isolated from the methanol extract of Z. nitidum var. fastuosum and elucidated by using spectroscopic techniques and comparison with the literature data. They were decarine (1) (Martin et al. 2005), norchelerythrine (2) (Jaromir et al. 2004), oxyavicine (3) (Hu et al. 2013), arnottianamide (4) (Hsiao & Chiang 1995), 6hydroxydihydrochelerythrine (5) (Seckarova et al. 2002), 6-methoxy-7-hydroxydihydrochelerythrine (6) (Tarus et al. 2006), 6-methoxydihydrochelerythrine (7) (Miao et al. 2011), 6carboxymethyldihydrochelerythrine (8) (Ng et al. 1987; Tsai et al. 2000), oxychelerythrine (9) (He et al. 2002), 6-acetonyldihydrochelerythrine (10) (Ye et al. 2009), 8-(10 -hydroxyethyl)-7,8dihydrochelerythrine (11) (Geng et al. 2009), N-nornitidine (12) (Martin et al. 2005) and noravicine (13) (Geen et al. 1998), and their structures are shown in Figure 1. According to the skeleton of the B-ring, these isolated compounds could be divided into unsaturation, fracture and saturation. All the compounds were isolated from Z. nitidum var. fastuosum for the first time.
Figure 1. The structures of compounds 1 – 13.
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Table 1. Cytotoxic activities of compounds 1 – 13. IC50(mM)a Compounds 1–5 6 7 8 9 10 11 12 13 Doxb
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a b
BALL-1
A549
HeLa
SMMC-7721
EJ
. 100 . 100 . 100 . 100 . 100 . 100 . 100 67.37 ^ 1.22 74.08 ^ 5.93 12.86 ^ 0.37
. 100 27.50 ^ 2.34 42.30 ^ 2.12 . 100 53.10 ^ 1.92 79.22 ^ 2.74 34.03 ^ 2.38 . 100 . 100 6.49 ^ 0.25
. 100 37.50 ^ 2.60 37.61 ^ 1.83 . 100 . 100 89.77 ^ 2.89 72.43 ^ 3.45 . 100 . 100 3.88 ^ 0.21
.100 16.95 ^ 0.89 17.93 ^ 1.56 .100 .100 .100 47.56 ^ 1.51 .100 .100 2.48 ^ 0.28
.100 60.42 ^ 3.00 70.81 ^ 3.33 .100 .100 .100 64.65 ^ 2.59 .100 .100 10.82 ^ 0.18
Inhibitory activity was expressed as the mean ^ SD of 50% inhibitory concentration of triplicate determinations. Doxorubicin was used as positive control.
The 13 isolates were evaluated preliminarily for cytotoxic activities against BALL-1, A549, Hela, SMMC-7721 and EJ tumour cell lines by the CCK-8 assay, with doxorubicin as the positive control. The results are presented in Table 1. In cell-proliferation activity, weak cytotoxic activities (IC50 . 100 mM) were observed in compounds 1– 5 and 8. Compound 6 exhibited the most potent cytotoxic activity against all cells except BALL-1, with IC50 values of 27.50, 37.50, 16.95 and 60.42 mM, respectively. Compound 7 showed similar cytotoxicity in comparison with compound 6. By comparing with compounds 6 and 7, compound 11 exhibited weaker cytotoxic activity against the above-mentioned four tumour cells with IC50 values of 40.79, 72.43, 47.56 and 68.29 mM, respectively, while compound 9 merely possessed cytotoxic effect against A549 with IC50 value of 53.10 mM. Among all the compounds, only 12 and 13 displayed cytotoxicity in inhibiting BALL-1 proliferation. The results revealed that the main cytotoxic activity of Z. nitidum var. fastuosum might be attributed to compounds 6, 7 and 9– 13. Some of the above-mentioned active compounds were also found to possess cytotoxicity in previous reports. For instance, compounds 5, 7 and 10 were tested against KB tumour cells in vitro (Stermitz et al. 1973); compounds 2, 9 and 10 exhibited cytotoxicity against P-388 or HT-29 cell lines (Chen et al. 2005). From the view of structure –activity relationship (Deng & Qin 2010), these in vitro results suggested that for the active compounds in this work, cytotoxic activities were enhanced by methoxylation at C-6. Indeed, compounds 6 and 7 showed stronger cytotoxicity against selected cells than other isolates, it seemed that the substitution at C-6 was more important than other positions in respect to the activity. However, BALL-1 was more susceptible to compounds 12 and 13, which might be associated with the unsaturated B-ring as well as substitution at C-8 and C-9. 3. Conclusions Z. nitidum var. fastuosum found in China primarily is the variation of Z. nitidum. This work was conducted for the purpose of investigating benzophenanthridine alkaloids from the roots of the titled plant. The isolation and cytotoxic activity of the 13 compounds provide more theoretical basis for broad-spectrum anti-cancer drug research. Supplementary material Experimental details relating to this paper are available online.
Natural Product Research
1383
Funding This project was supported by the Nuclear Accident Health Risk Assessment and Health Emergency Disposal Technology Research (2013BAK03B05).
Downloaded by [New York University] at 11:38 19 June 2015
References Chen JJ, Fang HY, Duh CY, Chen IS. 2005. New indolopyridoquinazoline, benzo[c ]phenanthridines and cytotoxic constituents from Zanthoxylum integrifoliolum. Planta Med. 71:470–475. Deng AJ, Qin HL. 2010. Cytotoxic dihydrobenzophenanthridine alkaloids from the roots of Macleaya microcarpa. Phytochemistry. 71:816–822. Geen GR, Mann IS, Mullane MV, McKillop A. 1998. A versatile synthesis of fully aromatic benzo[c ]phenanthridine alkaloids. Tetrahedron. 54:9875–9894. Geng D, Li DX, Shi Y, Liang JY, Min ZD. 2009. A new benzophenanthridine alkaloid from Zanthoxylum nitidum. Chin J Nat Med. 7:274–277. He WD, Van PL, De KN, Verbruggen L, Anthonissen K, Van der FM, Bosselaers J, Mathenge SG, Mudida FP. 2002. Chemical constituents and biological activities of Zanthoxylum usambarense. Phytother Res. 16:66–70. Hsiao JJ, Chiang HC. 1995. Lignans from the wood of Aralia bipinnata. Phytochemistry. 39:899–902. Hu J, Shi XD, Mao X, Chen JG, Zhu L, Zhao QJ. 2013. Antinociceptive activity of Rhoifoline A from the ethanol extract of Zanthoxylum nitidum in mice. J Ethanopharmacol. 150:828–834. Hu J, Zhang WD, Liu RH, Zhang C, Shen YH, Li HL, Liang MJ, Xu XK. 2006. Benzophenanthridine alkaloids from Zanthoxylum nitidum (Roxb.) DC, and their analgesic and anti-inflammatory activities. Chem Biodivers. 3:990–995. Hu K, Zhao SL, Ye FG, Lu X. 2008. Fingerprint analysis of Zanthoxylum nitidum by nonaqueous CE. Chromatographia. 68:475–479. Jaromir T, Jiri D, Radek M. 2004. Theoretical and experimental NMR chemical shifts of norsanguinarine and norchelerythrine. J Mol Struct. 689:115–120. Krane BD, Fagbule M, Shamma M, Gozler B. 1984. The benzophenanthridine alkaloids. J Nat Prod. 47:1–43. Luo X, Pedro L, Milic V, Mulhovo S, Duarte A, Duarte N, Ferreira MU. 2012. Antibacterial benzofuran neolignans and benzophenanthridine alkaloids from the roots of Zanthoxylum capense. Planta Med. 78:148–153. Mansoor TA, Borralho PM, Luo X, Mulhovo S, Rodrigues CMP, Ferreira MJU. 2013. Apoptosis inducing activity of benzophenanthridine-type alkaloids and 2-arylbenzofuran neolignans in HCT116 colon carcinoma cells. Phytomedicine. 20:923–929. Martin MT, Rasoanaivo LH, Raharisololalao A. 2005. Phenanthridine alkaloids from Zanthoxylum madagascariense. Fitoterapia. 76:590–593. Mester I, Waterman PG, Grundon MF. 1983. Chemistry and chemical taxonomy of the Rutales. London: academic Press. Miao F, Yang XJ, Zhou L, Hu HJ, Zheng F, Ding XD, Sun DM, Zhou CD, Sun W. 2011. Structural modification of sanguinarine and chelerythrine and their antibacterial activity. Nat Prod Res. 25:863–875. Ng KM, Gray AI, Waterman PG. 1987. Benzophenanthridine alkaloids from the stem bark of a Zanthoxylum specie. Phytochemistry. 26:3251–3254. Obiang-Obounou BW, Kang OH, Choi JG, Keum JH, Kim SB, Mun SH, Shin DW, Kim KW, Park CB, Kim YG. 2011. The mechanism of action of sanguinarine against methicillin-resistant Staphylococcus aureus. J Toxicol Sci. 36:277–283. Seckarova P, Marek R, Dostal J, Dommisse R, Esmans EL. 2002. Structural studies of benzophenanthridine alkaloid free bases by NMR spectroscopy. Magn Reson Chem. 40:147–152. Simanek V, Brossi A. 1985. The alkaloids, chemistry and pharmacology. Vol. 26. London: Academic Press; p. 186 –240. Stermitz FR, Larson KA, Kim DK. 1973. Some structural relationships among cytotoxic and antitumor benzophenanthridine alkaloid derivatives. J Med Chem. 16:939–940. Sun PW, Liu HT, Lei P, Li XZ, Huang Q, Li GY. 2011. RP-HPLC determination of nitidine chloride, ethoxychelerythrine and toddaloactone in Zanthoxylum nitidum (Roxb) DC. f. fastuosum How ex Huang. Cent S Pharm. 9:175–178. Tarus PK, Coombes PH, Crouch NR, Mulholland DA. 2006. Benzo[c ]phenanthridine alkaloids from stem bark of the Forest Knobwood, Zanthoxylum davyi (Rutaceae). S Afr J Bot. 72:555–558. Tsai IL, Ishikawa T, Seki H, Chen IS. 2000. Terihanine from Zanthoxylum nitidum. Chin Pharm J (Taipei). 52:43–49. Ueno T. 2008. Insecticides containing benzo[c ]phenanthridine alkaloids. Jpn Kokai Tokkyo Koho, JP 2008037775 A 20080221. Vanquelef E, Amoros M, Boustie J, Lynch MA, Waigh RD, Duval O. 2004. Synthesis and antiviral effect against herpes simplex type 1 of 12-substituted benzo[c ]phenanthridinium salts. J Enzym Inhib Med Chem. 19:481– 487. Wang HS, Ou SY, Pan YM, Chen ZF. 2006. Antibacterial activity and chemical constituents of the essential oils of Zanthoxylum nitidum var. fastuosum. Nat Prod Res Dev. 18:251–253. Ye FZ, Feng F, Liu WY. 2009. Alkaloids from Macleaya cordata. China J Chin Mater Med. 34:1683–1686.
Natural Product Research: Formerly Natural Product Letters Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/gnpl20
Cytotoxicity of benzophenanthridine alkaloids from the roots of Zanthoxylum nitidum (Roxb.) DC. var. fastuosum How ex Huang ab
a
a
b
Cheng-Fang Wang , Li Fan , Mei Tian , Shu-Shan Du , Zhi-Wei c
a
b
Deng , Jiang-Bin Feng , Yong-Yan Wang & Xu Su
a
a
Click for updates
China CDC Key Laboratory of Radiological Protection and Nuclear Emergency, National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention, Xicheng District, Beijing 100088, P.R. China b
State Key Laboratory of Earth Surface Processes and Resource Ecology, Beijing Normal University, No.19 Xinjiekouwai Street, Haidian District, Beijing 100875, P.R. China c
Analytic and Testing Center, Beijing Normal University, No.19 Xinjiekouwai Street, Haidian District, Beijing 100875, P.R. China Published online: 03 Feb 2015.
To cite this article: Cheng-Fang Wang, Li Fan, Mei Tian, Shu-Shan Du, Zhi-Wei Deng, Jiang-Bin Feng, Yong-Yan Wang & Xu Su (2015) Cytotoxicity of benzophenanthridine alkaloids from the roots of Zanthoxylum nitidum (Roxb.) DC. var. fastuosum How ex Huang, Natural Product Research: Formerly Natural Product Letters, 29:14, 1380-1383, DOI: 10.1080/14786419.2014.1002090 To link to this article: http://dx.doi.org/10.1080/14786419.2014.1002090
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or
howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content.
Downloaded by [New York University] at 11:38 19 June 2015
This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions
Natural Product Research, 2015 Vol. 29, No. 14, 1380–1383, http://dx.doi.org/10.1080/14786419.2014.1002090
SHORT COMMUNICATION Cytotoxicity of benzophenanthridine alkaloids from the roots of Zanthoxylum nitidum (Roxb.) DC. var. fastuosum How ex Huang Cheng-Fang Wangab, Li Fana, Mei Tiana, Shu-Shan Dub, Zhi-Wei Dengc, Jiang-Bin Fenga, Yong-Yan Wangb and Xu Sua*
Downloaded by [New York University] at 11:38 19 June 2015
a
China CDC Key Laboratory of Radiological Protection and Nuclear Emergency, National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention, Xicheng District, Beijing 100088, P.R. China; bState Key Laboratory of Earth Surface Processes and Resource Ecology, Beijing Normal University, No.19 Xinjiekouwai Street, Haidian District, Beijing 100875, P.R. China; cAnalytic and Testing Center, Beijing Normal University, No.19 Xinjiekouwai Street, Haidian District, Beijing 100875, P.R. China (Received 20 October 2014; final version received 18 December 2014)
This work aimed to investigate benzophenanthridine from the roots of Zanthoxylum nitidum (Roxb.) DC. var. fastuosum How ex Huang for the first time. Thirteen benzophenanthridines were isolated, and our results of the cytotoxic activities indicated that compound 6 exhibited the best potency against A549, Hela, SMMC-7721 and EJ, with the IC50 values of 27.50, 37.50, 16.95 and 60.42 mM, respectively. Compounds 7 and 11 also showed strong cytotoxicity when tested against the four human cancer cell lines (A549, Hela, SMMC-7721 and EJ), while only compounds 12 and 13 displayed cytotoxicity in inhibiting BALL-1 proliferation among all the compounds. These results suggested that benzophenanthridines may become a valid alternative of potential basis for new anti-proliferative agents. Keywords: benzophenanthridine alkaloids; Zanthoxylum nitidum (Roxb.) DC. var. fastuosum How ex Huang; cytotoxic activity
1. Introduction The genus Zanthoxylum is very well known for its diversified chemistry, particularly by the presence of alkaloids, aromatic and aliphatic amides, and phenylpropanoids – lignans and coumarins (Mester et al. 1983; Simanek & Brossi 1985). Among them, benzophenanthridine alkaloids are particularly abundant in roots and stem bark (Krane et al. 1984; Ng et al. 1987; Hu et al. 2006; Luo et al. 2012), which have shown broad spectrum of biological activities, including cytotoxic, antiviral, antibacterial, antinociceptive, anti-inflammatory and insecticidal properties (Vanquelef et al. 2004; Hu et al. 2006; Ueno 2008; Obiang-Obounou et al. 2011; Mansoor et al.
*Corresponding author. Email: [email protected] q 2015 Taylor & Francis
Natural Product Research
1381
Downloaded by [New York University] at 11:38 19 June 2015
2013). Zanthoxylum nitidum (Roxb.) DC. var. fastuosum How ex Huang (Z. nitidum var. fastuosum) is widely used to substitute Z. nitidum (Roxb.) DC. in folk medicine for the treatment of ulcer, toothache, stomachache and traumatic injuries, which was found in Guangxi province of China primarily (Wang et al. 2006). However, very few information on its chemical constituents and biological activities was reported in previous study (Wang et al. 2006; Hu et al. 2008; Sun et al. 2011). Further investigation is necessary to find the chemical basis of activities in this plant. In this work, we focus on benzophenanthridine alkaloids isolated from the roots of Z. nitidum var. fastuosum, aiming to identify the active compounds by assessing their cytotoxic activities against BALL-1, A549, Hela, SMMC-7721 and EJ cell lines in vitro. The correlation between structure and cytotoxicity is also discussed.
2. Results and discussion Thirteen benzophenanthridines were isolated from the methanol extract of Z. nitidum var. fastuosum and elucidated by using spectroscopic techniques and comparison with the literature data. They were decarine (1) (Martin et al. 2005), norchelerythrine (2) (Jaromir et al. 2004), oxyavicine (3) (Hu et al. 2013), arnottianamide (4) (Hsiao & Chiang 1995), 6hydroxydihydrochelerythrine (5) (Seckarova et al. 2002), 6-methoxy-7-hydroxydihydrochelerythrine (6) (Tarus et al. 2006), 6-methoxydihydrochelerythrine (7) (Miao et al. 2011), 6carboxymethyldihydrochelerythrine (8) (Ng et al. 1987; Tsai et al. 2000), oxychelerythrine (9) (He et al. 2002), 6-acetonyldihydrochelerythrine (10) (Ye et al. 2009), 8-(10 -hydroxyethyl)-7,8dihydrochelerythrine (11) (Geng et al. 2009), N-nornitidine (12) (Martin et al. 2005) and noravicine (13) (Geen et al. 1998), and their structures are shown in Figure 1. According to the skeleton of the B-ring, these isolated compounds could be divided into unsaturation, fracture and saturation. All the compounds were isolated from Z. nitidum var. fastuosum for the first time.
Figure 1. The structures of compounds 1 – 13.
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Table 1. Cytotoxic activities of compounds 1 – 13. IC50(mM)a Compounds 1–5 6 7 8 9 10 11 12 13 Doxb
Downloaded by [New York University] at 11:38 19 June 2015
a b
BALL-1
A549
HeLa
SMMC-7721
EJ
. 100 . 100 . 100 . 100 . 100 . 100 . 100 67.37 ^ 1.22 74.08 ^ 5.93 12.86 ^ 0.37
. 100 27.50 ^ 2.34 42.30 ^ 2.12 . 100 53.10 ^ 1.92 79.22 ^ 2.74 34.03 ^ 2.38 . 100 . 100 6.49 ^ 0.25
. 100 37.50 ^ 2.60 37.61 ^ 1.83 . 100 . 100 89.77 ^ 2.89 72.43 ^ 3.45 . 100 . 100 3.88 ^ 0.21
.100 16.95 ^ 0.89 17.93 ^ 1.56 .100 .100 .100 47.56 ^ 1.51 .100 .100 2.48 ^ 0.28
.100 60.42 ^ 3.00 70.81 ^ 3.33 .100 .100 .100 64.65 ^ 2.59 .100 .100 10.82 ^ 0.18
Inhibitory activity was expressed as the mean ^ SD of 50% inhibitory concentration of triplicate determinations. Doxorubicin was used as positive control.
The 13 isolates were evaluated preliminarily for cytotoxic activities against BALL-1, A549, Hela, SMMC-7721 and EJ tumour cell lines by the CCK-8 assay, with doxorubicin as the positive control. The results are presented in Table 1. In cell-proliferation activity, weak cytotoxic activities (IC50 . 100 mM) were observed in compounds 1– 5 and 8. Compound 6 exhibited the most potent cytotoxic activity against all cells except BALL-1, with IC50 values of 27.50, 37.50, 16.95 and 60.42 mM, respectively. Compound 7 showed similar cytotoxicity in comparison with compound 6. By comparing with compounds 6 and 7, compound 11 exhibited weaker cytotoxic activity against the above-mentioned four tumour cells with IC50 values of 40.79, 72.43, 47.56 and 68.29 mM, respectively, while compound 9 merely possessed cytotoxic effect against A549 with IC50 value of 53.10 mM. Among all the compounds, only 12 and 13 displayed cytotoxicity in inhibiting BALL-1 proliferation. The results revealed that the main cytotoxic activity of Z. nitidum var. fastuosum might be attributed to compounds 6, 7 and 9– 13. Some of the above-mentioned active compounds were also found to possess cytotoxicity in previous reports. For instance, compounds 5, 7 and 10 were tested against KB tumour cells in vitro (Stermitz et al. 1973); compounds 2, 9 and 10 exhibited cytotoxicity against P-388 or HT-29 cell lines (Chen et al. 2005). From the view of structure –activity relationship (Deng & Qin 2010), these in vitro results suggested that for the active compounds in this work, cytotoxic activities were enhanced by methoxylation at C-6. Indeed, compounds 6 and 7 showed stronger cytotoxicity against selected cells than other isolates, it seemed that the substitution at C-6 was more important than other positions in respect to the activity. However, BALL-1 was more susceptible to compounds 12 and 13, which might be associated with the unsaturated B-ring as well as substitution at C-8 and C-9. 3. Conclusions Z. nitidum var. fastuosum found in China primarily is the variation of Z. nitidum. This work was conducted for the purpose of investigating benzophenanthridine alkaloids from the roots of the titled plant. The isolation and cytotoxic activity of the 13 compounds provide more theoretical basis for broad-spectrum anti-cancer drug research. Supplementary material Experimental details relating to this paper are available online.
Natural Product Research
1383
Funding This project was supported by the Nuclear Accident Health Risk Assessment and Health Emergency Disposal Technology Research (2013BAK03B05).
Downloaded by [New York University] at 11:38 19 June 2015
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