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Cytotoxic activity of lignans from Hibiscus cannabinus
Fitoterapia 78 (2007) 385 – 387 www.elsevier.com/locate/fitote
Short report
Cytotoxic activity of lignans from Hibiscus cannabinus Laila Moujir a,⁎, Ana M.L. Seca b , Artur M.S. Silva c , Manuel R. López a , Nayely Padilla a , José A.S. Cavaleiro c , Carlos P. Neto c a
b
Department of Microbiology and Cellular Biology, and Instituto Canario de Investigación del Cáncer (ICIC), University of La Laguna, 38206 La Laguna, Tenerife, Spain Department of Technologic Sciences and Development, University of Azores, 9501-801 Ponta Delgada, Portugal c CICECO/QOPNA and Department of Chemistry, University of Aveiro, 3800-193 Aveiro, Portugal Received 7 April 2006; accepted 19 March 2007 Available online 11 April 2007
Abstract The antimicrobial and cytotoxic activities of six lignans isolated from the core and bark acetone extracts of Hibiscus cannabinus have been investigated. Two compounds (2 and 3) showed strong cytotoxic activity against HeLa, Hep-2 and A-549 cell lines while compound 5 showed moderate activity on HeLa cells when they were in advanced stage of cellular division. The compounds did not exhibit antimicrobial activity. © 2007 Published by Elsevier B.V. Keywords: Hibiscus cannabinus; Lignans; Cytotoxic activity; Antimicrobial activity
1. Plant Hibiscus cannabinus variety Salvador L. (Malvaceae), was harvested in an experimental farms in Quinta do Canal (latitude: 40°08′N, longitude 8°51′W, altitude 17 m), Figueira da Foz, Portugal, in September 1995. Authenticated by Dr. Ana M.L. Seca. 2. Uses in traditional medicine H. cannabinus is an annual dicotyledonous herbaceous plant known as “kenaf”. It is native of China but well known also in Africa and it has been recently cultivated in some Mediterranean areas as an alternative fiber crop for paper pulp production. It was used as antidote to poisoning with chemicals (acid, alkali, pesticides) and venomous mushrooms [1]. This plant is resistant to fungal pathogens [2], its essential oil has antifungal activity [3] and one of its constituents was found to be active against human cancer cell lines [4]. ⁎ Corresponding author. Tel.: +34 922318513; fax: +34 922318477. E-mail address: [email protected] (L. Moujir). 0367-326X/$ - see front matter © 2007 Published by Elsevier B.V. doi:10.1016/j.fitote.2007.03.010
386
L. Moujir et al. / Fitoterapia 78 (2007) 385–387
3. Previously isolated classes of constituents Lignans, lignanamides, tyramide derivatives and phenolic aldehydes [5,6], fatty acids, long chain alcohols, alkanes, sterols and triterpenes [7]. 4. Tested materials The lignans boehmenan K (1) (0.10%), boehmenan H (2) (0.11%), threo-carolignan K (3) (0.21%) and threocarolignan H (4) (0.05%) isolated from the acetone extract of core and grosamide K (5) (0.25%) and erythro-canabisine H (6) (0.42%) from the acetone extract of bark (Fig. 1). 5. Studied activity HeLa (human carcinoma of the cervix), Hep-2 (human carcinoma of the larynx) and A-549 (human lung carcinoma) cell lines were grown as a monolayer in DMEM, supplemented with 10% fetal bovine serum and 1% of penicillin– streptomycin mixture (10,000 UI/ml). Cytotoxicity was assessed using the colorimetric MTT reduction assay as previously reported [8]. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were determined by the broth microdilution method in 96-well microtiter plates [9]. 6. Used microorganisms Staphylococcus aureus ATCC 6538, S. epidermidis ATCC 14990, S. saprophyticus ATCC 15305, Bacillus subtilis ATCC 6051, B. pumilus ATCC 7061, B. cereus ATCC 21776, B. megaterium ATCC 25848, Enterococcus faecalis ATCC 29212, Mycobacterium smegmati ATCC 19420. Escherichia coli ATCC 9637, Pseudomonas aeruginosa AK
Fig. 1. Structures of compounds 1–6.
L. Moujir et al. / Fitoterapia 78 (2007) 385–387
387
Table 1 Cytotoxicity of compounds 1–6 against HeLa, Hep-2 and A-549 cells Compounds
IC50 (μg/ml) HeLa
1 2 3 4 5 6 Actinomycin Da
Hep-2
A549
lag phase
log phase
lag phase
log phase
lag phase
log phase
N20 2.6 1 N20 N20 N20 0.014
N20 1.7 0.6 N20 17.2 N20 0.002
N20 N20 7.2 N20 N20 N20 0.24
N20 10.7 3.3 N20 N20 N20 0.008
N20 14.5 9.5 N20 N20 N20 NA
N20 4.2 3.7 N20 N20 N20 NA
a
Actinomycin D was used as a positive control. NA: Not assayed.
958 (from the British Columbia University), Proteus mirabilis CETC 170, Salmonella sp. CECT 456 and Candida albicans UBC 1 (from the British Columbia University). 7. Results The cytotoxic activity results are reported in Table 1. The MIC and MBC for all the tested compounds are N 40 μg/ml. 8. Conclusions The cytotoxic activity data (Table 1) showed that boehmenan H (2) and threo-carolignan K (3) were the most active compounds under the two conditions assayed and against the three tested cell lines. It is important to emphasize that when the cells are exponentially grown, the activity increases considerably in both compounds. On the basis of their chemical features the results suggest that the loss of dihydrofuran system conjunctly with the presence of feruloyl moiety increase the activity significantly against the cell lines used (3 versus 1, 2 and 4). Furthermore, the tyramine group together with the benzodihydrofuran system increased slightly the cytotoxic activity (5 versus 6) but only against HeLa cells. All the assayed compounds were inactive against the microorganism used (MIC N 40 μg/ml). Acknowledgements Thanks are due to the University of Aveiro and FCT-Lisbon for funding the Research Units QOPNA and CICECO. Ana M.L. Seca is grateful to PRAXIS XXI for a Ph.D. grant (BD/3412/94). Spanish Grants BQU2003-09558-CO2-01 and ICIC/G.I (01/05). References [1] [2] [3] [4] [5] [6] [7] [8] [9]
Chifundera K, Balagizi K, Kizungu B. Fitoterapia 1994;65:307. Pshenichnov EA, Sultanova EM, Veshkurova ON, Ionov MV, Salakhutdinov BA, Salikhov SI. Chem Nat Comp 2004;40:63. Kobaisy M, Tellez MR, Webber CL, Dayan FE, Schader KK, Wedge DE. J Agric Food Chem 2001;49:3768. Chin Y-W, Jones WP, Rachman I, Riswan S, Kardono LBS, Chai H-B, et al. Phytother Res 2006;20:62. Seca AML, Silva AMS, Silvestre AJD, Cavaleiro JAS, Domingues FMJ, Pascoal-Neto C. Phytochemistry 2001;58:1219. Seca AML, Silva AMS, Silvestre AJD, Cavaleiro JAS, Domingues FMJ, Pascoal-Neto C. Phytochemistry 2001;56:759. Seca AML, Silva AMS, Silvestre AJD, Cavaleiro JAS, Domingues FMJ, Pascoal-Neto C. Phytochem Anal 2000;11:345. Dias T, Brito I, Moujir L, Paiz N, Darias J, Cueto M. J Nat Prod 2005;68:1677. De León L, Beltrán B, Moujir L. Planta Med 2005;71:313.
Short report
Cytotoxic activity of lignans from Hibiscus cannabinus Laila Moujir a,⁎, Ana M.L. Seca b , Artur M.S. Silva c , Manuel R. López a , Nayely Padilla a , José A.S. Cavaleiro c , Carlos P. Neto c a
b
Department of Microbiology and Cellular Biology, and Instituto Canario de Investigación del Cáncer (ICIC), University of La Laguna, 38206 La Laguna, Tenerife, Spain Department of Technologic Sciences and Development, University of Azores, 9501-801 Ponta Delgada, Portugal c CICECO/QOPNA and Department of Chemistry, University of Aveiro, 3800-193 Aveiro, Portugal Received 7 April 2006; accepted 19 March 2007 Available online 11 April 2007
Abstract The antimicrobial and cytotoxic activities of six lignans isolated from the core and bark acetone extracts of Hibiscus cannabinus have been investigated. Two compounds (2 and 3) showed strong cytotoxic activity against HeLa, Hep-2 and A-549 cell lines while compound 5 showed moderate activity on HeLa cells when they were in advanced stage of cellular division. The compounds did not exhibit antimicrobial activity. © 2007 Published by Elsevier B.V. Keywords: Hibiscus cannabinus; Lignans; Cytotoxic activity; Antimicrobial activity
1. Plant Hibiscus cannabinus variety Salvador L. (Malvaceae), was harvested in an experimental farms in Quinta do Canal (latitude: 40°08′N, longitude 8°51′W, altitude 17 m), Figueira da Foz, Portugal, in September 1995. Authenticated by Dr. Ana M.L. Seca. 2. Uses in traditional medicine H. cannabinus is an annual dicotyledonous herbaceous plant known as “kenaf”. It is native of China but well known also in Africa and it has been recently cultivated in some Mediterranean areas as an alternative fiber crop for paper pulp production. It was used as antidote to poisoning with chemicals (acid, alkali, pesticides) and venomous mushrooms [1]. This plant is resistant to fungal pathogens [2], its essential oil has antifungal activity [3] and one of its constituents was found to be active against human cancer cell lines [4]. ⁎ Corresponding author. Tel.: +34 922318513; fax: +34 922318477. E-mail address: [email protected] (L. Moujir). 0367-326X/$ - see front matter © 2007 Published by Elsevier B.V. doi:10.1016/j.fitote.2007.03.010
386
L. Moujir et al. / Fitoterapia 78 (2007) 385–387
3. Previously isolated classes of constituents Lignans, lignanamides, tyramide derivatives and phenolic aldehydes [5,6], fatty acids, long chain alcohols, alkanes, sterols and triterpenes [7]. 4. Tested materials The lignans boehmenan K (1) (0.10%), boehmenan H (2) (0.11%), threo-carolignan K (3) (0.21%) and threocarolignan H (4) (0.05%) isolated from the acetone extract of core and grosamide K (5) (0.25%) and erythro-canabisine H (6) (0.42%) from the acetone extract of bark (Fig. 1). 5. Studied activity HeLa (human carcinoma of the cervix), Hep-2 (human carcinoma of the larynx) and A-549 (human lung carcinoma) cell lines were grown as a monolayer in DMEM, supplemented with 10% fetal bovine serum and 1% of penicillin– streptomycin mixture (10,000 UI/ml). Cytotoxicity was assessed using the colorimetric MTT reduction assay as previously reported [8]. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were determined by the broth microdilution method in 96-well microtiter plates [9]. 6. Used microorganisms Staphylococcus aureus ATCC 6538, S. epidermidis ATCC 14990, S. saprophyticus ATCC 15305, Bacillus subtilis ATCC 6051, B. pumilus ATCC 7061, B. cereus ATCC 21776, B. megaterium ATCC 25848, Enterococcus faecalis ATCC 29212, Mycobacterium smegmati ATCC 19420. Escherichia coli ATCC 9637, Pseudomonas aeruginosa AK
Fig. 1. Structures of compounds 1–6.
L. Moujir et al. / Fitoterapia 78 (2007) 385–387
387
Table 1 Cytotoxicity of compounds 1–6 against HeLa, Hep-2 and A-549 cells Compounds
IC50 (μg/ml) HeLa
1 2 3 4 5 6 Actinomycin Da
Hep-2
A549
lag phase
log phase
lag phase
log phase
lag phase
log phase
N20 2.6 1 N20 N20 N20 0.014
N20 1.7 0.6 N20 17.2 N20 0.002
N20 N20 7.2 N20 N20 N20 0.24
N20 10.7 3.3 N20 N20 N20 0.008
N20 14.5 9.5 N20 N20 N20 NA
N20 4.2 3.7 N20 N20 N20 NA
a
Actinomycin D was used as a positive control. NA: Not assayed.
958 (from the British Columbia University), Proteus mirabilis CETC 170, Salmonella sp. CECT 456 and Candida albicans UBC 1 (from the British Columbia University). 7. Results The cytotoxic activity results are reported in Table 1. The MIC and MBC for all the tested compounds are N 40 μg/ml. 8. Conclusions The cytotoxic activity data (Table 1) showed that boehmenan H (2) and threo-carolignan K (3) were the most active compounds under the two conditions assayed and against the three tested cell lines. It is important to emphasize that when the cells are exponentially grown, the activity increases considerably in both compounds. On the basis of their chemical features the results suggest that the loss of dihydrofuran system conjunctly with the presence of feruloyl moiety increase the activity significantly against the cell lines used (3 versus 1, 2 and 4). Furthermore, the tyramine group together with the benzodihydrofuran system increased slightly the cytotoxic activity (5 versus 6) but only against HeLa cells. All the assayed compounds were inactive against the microorganism used (MIC N 40 μg/ml). Acknowledgements Thanks are due to the University of Aveiro and FCT-Lisbon for funding the Research Units QOPNA and CICECO. Ana M.L. Seca is grateful to PRAXIS XXI for a Ph.D. grant (BD/3412/94). Spanish Grants BQU2003-09558-CO2-01 and ICIC/G.I (01/05). References [1] [2] [3] [4] [5] [6] [7] [8] [9]
Chifundera K, Balagizi K, Kizungu B. Fitoterapia 1994;65:307. Pshenichnov EA, Sultanova EM, Veshkurova ON, Ionov MV, Salakhutdinov BA, Salikhov SI. Chem Nat Comp 2004;40:63. Kobaisy M, Tellez MR, Webber CL, Dayan FE, Schader KK, Wedge DE. J Agric Food Chem 2001;49:3768. Chin Y-W, Jones WP, Rachman I, Riswan S, Kardono LBS, Chai H-B, et al. Phytother Res 2006;20:62. Seca AML, Silva AMS, Silvestre AJD, Cavaleiro JAS, Domingues FMJ, Pascoal-Neto C. Phytochemistry 2001;58:1219. Seca AML, Silva AMS, Silvestre AJD, Cavaleiro JAS, Domingues FMJ, Pascoal-Neto C. Phytochemistry 2001;56:759. Seca AML, Silva AMS, Silvestre AJD, Cavaleiro JAS, Domingues FMJ, Pascoal-Neto C. Phytochem Anal 2000;11:345. Dias T, Brito I, Moujir L, Paiz N, Darias J, Cueto M. J Nat Prod 2005;68:1677. De León L, Beltrán B, Moujir L. Planta Med 2005;71:313.