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Chemical constituents isolated from the bark of Guatteria blepharophylla (Annonaceae) and their antiproliferative and antimicrobial activities

Abstracts

Phytochemical study of the bark of Guatteria blepharophylla (Mart.) Mart. afforded twelve compounds, namely two sesquiterpenes, caryophyllene oxide (1) and spathulenol (3), one xanthone, lichexanthone (2), a mixture of steroids, β-sitosterol (4), and stigmasterol (5), and seven isoquinoline alkaloids, O-methylmoschatoline (6), lysicamine (7), nornuciferine (8), liriodenine (9), isocoreximine (10), subsessiline (11), and isomoschatoline (12). Their structures were established on the basis of spectroscopic methods. Compounds 1-6, 11 and 12 were reported for the first time in this species. The 13C NMR (nuclear magnetic resonance) data for the compounds 11 and 12 are described for the first time in the literature. The antiproliferative activity against human tumour cell lines and antimicrobial activities were investigated for the major compounds. Compound 9 showed significant activity against cell lines of breast (MCF-7, Michigan Cancer Foundation-7), superior to the positive control doxorubicin. Compound 12 presented antifungal activity similar to the positive control nystatin against Candida albicans.

Guatteria blepharophylla; antifungal and antiproliferative activities; alkaloids; terpenes; xanthone


O estudo fitoquímico das cascas de Guatteria blepharophylla (Mart.) Mart. forneceu 12 compostos, sendo dois sesquiterpenos, óxido de cariofileno (1) e espatulenol (3), uma xantona, liquexantona (2), uma mistura de esteróides, β-sitosterol (4) e estigmasterol (5), e sete alcalóides isoquinolínicos, O-metilmoschatolina (6), lysicamina (7), nornuciferina (8), liriodenina (9), isocoreximina (10), subsessilina (11) e isomoschatolina (12). Suas estruturas foram determinadas através de métodos espectroscópicos. Os compostos 1-6, 11 e 12 são reportados pela primeira vez nesta espécie. Os dados de RMN 13C (ressonância magnética nuclear) para os compostos 11 e 12 são descritos pela primeira vez na literatura. As atividades antiproliferativa em linhagens de células tumorais humanas e antimicrobiana foram investigadas para os compostos majoritários. O composto 9 mostrou significativa atividade contra linhagens de células de mama (MCF-7, Michigan Cancer Foundation-7), superior ao controle positivo doxorrubicina. O composto 12 apresentou atividade antifúngica similar ao controle positivo nistatina contra Candida albicans


ARTICLE

Chemical constituents isolated from the bark of Guatteria blepharophylla (Annonaceae) and their antiproliferative and antimicrobial activities

Emmanoel V. CostaI,II; Francisco de Assis MarquesI; Maria Lúcia B. PinheiroIII; Raquel M. BragaIV; Camila DelarmelinaV; Marta Cristina T. DuarteV; Ana Lúcia T. G. RuizV; João Ernesto de CarvalhoV; Beatriz H. L. N. S. MaiaI,* * e-mail: noronha@ufpr.br

IDepartamento de Química, Universidade Federal do Paraná, CP 19081, 81531-990 Curitiba-PR, Brazil

IIDepartamento de Química, Universidade Federal de Sergipe, 49100-000 São Cristovão-SE, Brazil

IIIDepartamento de Química, Universidade Federal do Amazonas, 69077-000 Manaus-AM, Brazil

IVInstituto de Química, Universidade Estadual de Campinas, CP 6154, 13083-970 Campinas-SP, Brazil

VDivisão de Microbiologia and Divisão de Farmacologia e Toxicologia, CPQBA, Universidade Estadual de Campinas, CP 6171, 13083-970 Campinas-SP, Brazil

ABSTRACT

Phytochemical study of the bark of Guatteria blepharophylla (Mart.) Mart. afforded twelve compounds, namely two sesquiterpenes, caryophyllene oxide (1) and spathulenol (3), one xanthone, lichexanthone (2), a mixture of steroids, β-sitosterol (4), and stigmasterol (5), and seven isoquinoline alkaloids, O-methylmoschatoline (6), lysicamine (7), nornuciferine (8), liriodenine (9), isocoreximine (10), subsessiline (11), and isomoschatoline (12). Their structures were established on the basis of spectroscopic methods. Compounds 1-6, 11 and 12 were reported for the first time in this species. The 13C NMR (nuclear magnetic resonance) data for the compounds 11 and 12 are described for the first time in the literature. The antiproliferative activity against human tumour cell lines and antimicrobial activities were investigated for the major compounds. Compound 9 showed significant activity against cell lines of breast (MCF-7, Michigan Cancer Foundation-7), superior to the positive control doxorubicin. Compound 12 presented antifungal activity similar to the positive control nystatin against Candida albicans.

Keywords: Guatteria blepharophylla, antifungal and antiproliferative activities, alkaloids, terpenes, xanthone

RESUMO

O estudo fitoquímico das cascas de Guatteria blepharophylla (Mart.) Mart. forneceu 12 compostos, sendo dois sesquiterpenos, óxido de cariofileno (1) e espatulenol (3), uma xantona, liquexantona (2), uma mistura de esteróides, β-sitosterol (4) e estigmasterol (5), e sete alcalóides isoquinolínicos, O-metilmoschatolina (6), lysicamina (7), nornuciferina (8), liriodenina (9), isocoreximina (10), subsessilina (11) e isomoschatolina (12). Suas estruturas foram determinadas através de métodos espectroscópicos. Os compostos 1-6, 11 e 12 são reportados pela primeira vez nesta espécie. Os dados de RMN 13C (ressonância magnética nuclear) para os compostos 11 e 12 são descritos pela primeira vez na literatura. As atividades antiproliferativa em linhagens de células tumorais humanas e antimicrobiana foram investigadas para os compostos majoritários. O composto 9 mostrou significativa atividade contra linhagens de células de mama (MCF-7, Michigan Cancer Foundation-7), superior ao controle positivo doxorrubicina. O composto 12 apresentou atividade antifúngica similar ao controle positivo nistatina contra Candida albicans.

Introduction

The genus Guatteria Ruiz & Pav. contains close to 290 species and is the largest genus within the Annonaceae family.1 Species of Guatteria are frequent constituents of Neotropical (lowland) forests, and it is widely distributed throughout Mesoamerica, the Caribbean and tropical South America.1

Plants of this family are known for their edible fruits and medicinal properties of many species.2 Previous chemical and pharmacological investigations on some species of this family, including Guatteria, have indicated the presence of important bioactive compounds, exhibiting many pharmacological activities, such as, cytotoxicity against human tumor cell lines,3-5 antimicrobial,5,6-9 and antiparasitic properties, particularly against Leishmania sp.,3,7,10-12 Plasmodium falciparum 5,12,13 and Trypanosoma sp.3,12,14

Despite the importance of annonaceous members in folk medicine, the number of species that have been chemically investigated is still very small. One of them is Guatteria blepharophylla Mart in Mart, a small tropical tree that occurs in the Amazonian region (Brazil, Peru, Guyana, Ecuador and Venezuela).1 In Brazil this species is common in Amazonas and Pará states, where it is known as "envireira".15 Previous phytochemical studies on this species described the isolation and identification of essential oils and isoquinoline alkaloids.8,15,16

In continuation of our research on bioactive compounds from Amazonian annonaceous plants, we report herein the phytochemical study of the bark of G. blepharophylla and the evaluation of antiproliferative and antimicrobial properties of the main isolated compounds.

Experimental

General experimental procedures

Melting points were determined on a Quimis Q-340S23 micromelting point apparatus. UV-Vis spectra were obtained in CH3OH on a Hewlett-Packard HP 8452A diode array spectrophotometer. IR spectra were acquired on a BIORAD FTS-3500 GX spectrophotometer. Mass spectra were recorded on a Varian Saturn 2000 spectrometer operating at 70 eV. NMR data, 1D and 2D, were recorded at 293 K in CDCl3 or CDCl3 + CD3OD or CD3OD on a Bruker Avance DRX 400 and Brucker ARX 200 spectrometers. The spectrometers were equipped with a 5 mm multinuclear direct detection probe with z-gradient. One-bond and long-range 1H-13C correlation (HSQC, (heteronuclear single quantum correlation) and HMBC (heteronuclear multiple bond coherence), respectively) experiments were optimized for an average coupling constants 1J(C,H) and LRJ(C,H) of 140 and 8 Hz, respectively. All 1H and 13C NMR chemical shifts (σ) are given in ppm related to the TMS (tetramethylsilane) signal at 0.00 ppm as internal reference and the coupling constants (J) in Hz. Silica gel 60 (70-230 mesh) was used for column chromatography, while silica gel 60 F254 were used for analytical (0.25 mm), and preparative (1.00 mm) TLC. Compounds were visualized by exposure under UV254/366 light, spraying p-anisaldeyde reagent followed by heating on a hot plate, or spraying with Dragendorff's reagent.

Plant material

The bark of G. blepharophylla was collected in January 2005 on the campus of the Universidade Federal do Amazonas (UFAM), Manaus-AM, Brazil, and identified by the taxonomist Prof. Dr. A. C. Webber from UFAM. A voucher specimen (number 7340) has been deposited at the Herbarium of the UFAM. After identification, G. blepharophylla bark was dried at room temperature and finely powdered.

Extraction and isolation of the chemical constituents

The dried powdered bark (1500 g) of G. blepharophylla was successively extracted with n-hexane followed by MeOH to yield n-hexane (28.18 g) and MeOH (212.50 g) extracts.

The n-hexane extract (3.50 g) was subjected to silica gel column chromatography eluted with the gradient systems: petroleum ether-CH2Cl2 from 100:0 to 10:90 (v/v) followed by CH2Cl2-EtOAc from 100:0 to 10:90 (v/v), and EtOAc-MeOH from 100:0 to 80:20 (v/v), affording 295 fractions (each 15 mL). The eluted fractions were evaluated and pooled according to TLC analysis, to afford 24 groups of fractions. Fraction 4 (340.7 mg) was submitted to further silica gel column chromatography eluted with the gradient systems: petroleum ether-CH2Cl2 from 100:0 to 10:90 (v/v), and CH2Cl2-EtOAc from 100:0 to 10:90 (v/v), yielding 40 subfractions that were pooled in nine subfractions according to TLC analysis. Subfraction 4.4 (154.7 mg) was purified by preparative TLC eluted with petroleum ether-EtOAc (90:10, v/v, two times) to give 1 (56.7 mg) and 2 (20.0 mg). Fraction 5 (187.2 mg) was also submitted to another silica gel column chromatography using the same methodology described for fraction 4 affording 40 new subsfractions that were pooled in nine groups of subfractions, according to TLC analysis. Subfractions 5.3 (14.0 mg) and 5.4 (55.7 mg) were purified by preparative TLC eluted with petroleum ether-EtOAc (90:10, v/v, two times) to give 2 (3.0 mg), and 3 (16.8 mg), respectively. Fraction 6 were also submitted to silica gel column chromatography using the same methodology above yielding 56 subfractions that were pooled in ten groups of subfractions, according to TLC analysis. Subfraction 6.6 was purified by preparative TLC eluted with petroleum ether-EtOAc (80:20, v/v, two times) to yield a mixture of 4 and 5 (36.1 mg).

TLC investigations indicated a high concentration of alkaloids in the MeOH extract. Therefore, an aliquot of the MeOH extract (210.0 g) was initially subjected to an acid-base extraction10 to give the CH2Cl2 alkaloid fraction (1.18 g) and the CH2Cl2 neutral fraction (5.24 g). The alkaloid fraction (1.0 g) was subjected to 10% NaHCO3 treated silica gel column chromatography10 eluted with the gradient systems: petroleum ether-CH2Cl2 from 100:0 to 10:90 (v/v) followed by CH2Cl2-EtOAc from 100:0 to 10:90 (v/v), and EtOAc-MeOH from 100:0 to 50:50 (v/v), yielding 97 fractions (each 25 mL). The eluted fractions were evaluated and pooled according to TLC analysis, to afford 13 groups of fractions. Fraction 4 (112.8 mg) was purified by preparative TLC eluted with CH2Cl2-MeOH (95:05, v/v, two times) to give 6 (13.0 mg), 7 (7.0 mg), and 8 (8.0 mg). Fraction 5 (87.0 mg) was also purified by preparative TLC eluted with CH2Cl2-MeOH (95:05, v/v, two times) to yield 7 (12.6 mg), 8 (2.8 mg) and 9 (2.0 mg). Fraction 6 (282.3 mg) was also submitted to another silica gel column chromatography using the same methodology above affording 27 new subsfractions that were pooled in six groups of subfractions, according to TLC analysis. Subfraction 6.3 (101.5 mg) was purified by preparative TLC eluted with CH2Cl2-MeOH (90:10, v/v, two times) to give 10 (9.0 mg). Fraction 8 was purified by preparative TLC eluted with CH2Cl2-MeOH (95:05, v/v, three times) to give 11 (12.7 mg). Fraction 10 was also purified by preparative TLC eluted with CH2Cl2-MeOH (90:10, v/v, three times) to give 12 (31.2 mg).

Subsessiline ( 11 )

Orange needles (CHCl3:MeOH 2:1); UV λmax/nm (MeOH) (log e) 204 (3.89), 238 (3.41), 278 (3.54), 472 (2.95); IR νmax/cm-1 (KBr) 3431, 3132, 3097, 2955, 2934, 2855, 1646, 1602, 1582, 1543, 1492, 1466, 1394, 1380, 1326, 1258, 1206, 1087, 1055, 1015, 994, 872; 1H and 13C NMR data: Table 2.

Isomoschatoline ( 12 )

Blue needles (CHCl3:MeOH 2:1); UV λmax/nm (MeOH) (log e) 204 (4.23), 238 (3.84), 282 (4.01), 304 (3.93), 468 (3.15), 595 (3.09); IR νmax/cm-1 (KBr) 3325, 2938, 2843, 1660, 1614, 1596, 1578, 1541, 1494, 1410, 1367, 1337, 1300, 1266, 1202, 1042, 996, 801, 691; 1H and 13C NMR data: Table 2.

In vitro anticancer activity assay

Human tumour cell lines UACC-62 (melanoma), MCF7 (breast), NCI-H460 (lung, non-small cells), OVCAR-03 (ovarian), PC-3 (prostate), HT-29 (colon), 786-0 (renal), K562 (leukemia) and NCI-ADR/RES (ovarian expressing phenotype multiple drugs resistance) were kindly provided by National Cancer Institute (NCI). Stock cultures were grown in medium containing 5 mL RPMI 1640 (GIBCO® BRL) supplemented with 5% fetal bovine serum.

Penicillin:streptomycin (1000 μg mL-1:1000 UI mL-1 , 1 mL L-1) was added to experimental cultures. Cells in 96 well plates (100 μL cells well-1) were exposed to sample concentrations in DMSO/RPMI (0.25, 2.5, 25 and 250 μg mL-1) (DMSO, dimethyl sulfoxide) at 37 ºC, 5% of CO2 in air for 48 h. Final DMSO concentration did not affect cell viability. Afterwards, cells were fixed with 50% trichloroacetic acid and cell proliferation determined by spectrophotometric quantification (540 nm) of cellular protein content using sulforhodamine B assay. Using the concentration-response curve for each cell line, TGI (concentration that produces total growth inhibition or cytostatic effect)17 was determined through non-linear regression analysis (Table 1) using software ORIGIN 7.5 (OriginLab Corporation).

In vitro antimicrobial activity

The growth inhibitory activity of the crude extracts, fractions and isolated compounds was tested against 11 microorganisms (Bacillus subtilis ATCC 5061, Candida albicans ATCC 10231, Enterococcus faecium CCT 5079, Enterococcus hirae ATCC 10541, Escherichia coli ATCC 11775, Micrococcus luteus ATCC 4698, Pseudomonas aeruginosa ATCC 13388, Rhodococcus equi ATCC 6939, Salmonella choleraesuis ATCC 10708, Staphylococcus aureus ATCC 6538 and Staphylococcus epidermidis ATCC 12228).

The bacteria strains were cultured overnight at 36 ºC in Nutrient Agar (Merck), while C. albicans was cultured in Saboraud Dextrose Agar. Inoculum for the assays was prepared by diluting a scraped cell mass in 0.85% NaCl solution, adjusted to McFarland scale 0.5 and confirmed by spectrophotometer reading at 580 nm. Cell suspensions were finally diluted to 104 CFU mL-1 for using in the activity assays. MIC (minimal inhibitory concentration) tests were carried out according to Eloff,18 using Müller-Hinton broth on a tissue-culture test plate (96 wells). The stock solution crude extracts, fractions and isolated compounds were diluted and transferred into the first well, and serial dilutions were made so that concentrations in the range of 1.0-0.015 mg mL-1 were obtained. Chloramphenicol and nystatin (Merck) were used as the reference antibiotic control in the range of 0.25-0.002 mg mL-1. The inoculum was added to all wells, and the plates were incubated at 36 ºC for 48 h. Each concentration was screened in triplicate. Antimicrobial activity was detected by adding 20 μL of 0.5% TTC (triphenyl tetrazolium chloride, Merck) aqueous solution. MIC was defined as the lowest concentration of the sample that inhibited visible growth, as indicated by TCC staining (dead cells are not stained by TTC).

Results and Discussion

Once n-hexane and MeOH crude extracts were found to have antiproliferative activity (Table 1), these extracts were subjected to successive chromatographic fractionations as described in the Experimental section leading to the isolation of the chemical constituents 1-12 (Figure 1).


Compounds 1-10 were identified by comparison of their spectroscopic data with those reported in the literature as caryophyllene oxide19 (1), lichexanthone20 (2), spathulenol19 (3), a mixture of β-sitosterol21 (4), and stigmasterol21 (5), O-methylmoschatoline22,23 (6), lysicamine22,23 (7), nornuciferine22 (8), liriodenine6,23 (9), and isocoreximine15 (10). Compounds 7-10 were recently found on the stem of this species,15 while compounds 1-6, 11 and 12 were reported for the first time in this specie. Compound 2 is related for the first time in the genus Guatteria, and it is the second report in the Annonaceae, previously isolated from the roots of Rollinia leptopetala.24

Compound 11 was obtained as an orange amorphous powder which was positive to the Dragendorff's test. Its UV-vis spectrum showed absorption peaks at 204, 238, 278 and 472 nm, which are typical of molecules with an oxoaporphine skeleton. The IR spectrum showed absorption bands at 3431 and 1646 cm-1, characteristic of phenolic hydroxyl and carbonyl groups. The 1H NMR spectrum indicated a tetrasubstituted-oxoaporphine alkaloid. Its 1H NMR spectrum revealed the presence of a pair of doublets at σ 8.88 and σ 8.24 (H-5 and H-4, J 4.7 Hz, respectively), which was consistent with the presence of a pyridine system. Additionally, it was found to have a spin system consisting of three hydrogens with the signals at σ 8.96 (1H, d, J 9.0 Hz), σ 7.82 (1H, d, J 2.8 Hz), and σ 7.25 (1H, dd, J 9.0 and 2.8 Hz), which were attributed to H-11, H-8, and H-10, respectively, and indicated substitution in the D ring. The signals at σ 4.05, σ 4.11 and σ 4.17 (each 3H, s) were assigned to three methoxyl groups located in the A ring, according to long-range 1H-13C correlation in the HMBC NMR experiment (Table 2). The presence of a hydroxyl group in the molecule located in the D ring at C-9 was established on the basis of the long-range 1H-13C correlation of the hydrogen H-11 at σ 8.96 with the carbon C-9 at σ 157.3, which showed no correlation with the methoxyl groups.

The 13C NMR experiment along with HSQC and HMBC experiments allowed to verify the presence of 19 carbons, including a carbonyl group at σ 183.1, 15 aromatic carbons between σ 157.3-113.4, and three methoxyl groups at σ 61.9, σ 61.4, and σ 60.9, which was in agreement with structure 11 (Table 2). The hydrogen H-8 at σ 7.82 showed strong long-range 1H-13C correlation with the carbon C-7 at σ 183.1, which confirmed the oxoaporphine skeleton. According with 1H and 13C NMR 1D/2D this compound was identified as the oxoaporphine alkaloid known subsessiline. This compound has been found only in two species of Annonaceae Guatteria ouregou25 and G. subsessilis,26 but the physical data are incomplete. The 13C NMR data are reported for the first time. The complete physical data for this compound are described in this work.

Compound 12 was obtained as a blue amorphous powder. Its UV-Vis spectrum showed absorption peaks at 204, 238, 282, 304, 468 and 595 nm, which are typical of molecules with an oxoaporphine skeleton. The IR spectrum showed absorption bands at 3325 and 1660 cm-1, characteristic of phenolic hydroxyl and carbonyl groups. The 1H and 13C NMR spectra of 12 were very similar to those of 11, except for the absence of a methoxyl signal, which was replaced by a hydroxyl group, and absence of substitution in the D ring, in structure 12 according to the IR spectrum and NMR data (Table 2). The absence of substitution in the D ring was confirmed by the presence of four adjacent aromatic hydrogens at σ 9.08 (1H, ddd, J 8.6, 1.0 and 0.5 Hz), σ 8.41 (1H, ddd, J 8.0, 1.6 and 0.5 Hz), σ 7.68 (1H, ddd, J 8.6, 7.0 and 1.6 Hz), and σ 7.33 (1H, ddd, J 8.0, 7,0 and 1.0 Hz) which were attributed to H-11, H-8, H-10, and H-9, respectively (Table 2). The position of the hydroxyl group at C-3 was established on the basis of the long-range 1H-13C correlation of the hydrogen H-4 at σ 8.60 with the carbon C-3 at σ 166.0, which showed no correlation with the methoxyl groups. According with 1H and 13C NMR 1D/2D, this compound was identified as the oxoaporphine alkaloid known isomoschatoline. As well as observed for 11, the physical data published for this compound are incomplete and presented in this work. The 13C NMR data is reported for the first time. This compound has been found only in three species ofAnnonaceae Uvaria mocoli,27 Guatteria melosma and Cleistopholis patens.28

The isolated compounds 1-2 and 6-12 were evaluated for in vitro antiproliferative activity against nine human tumour cell lines (Table 1), while compounds 6, 7, 10 and 12 were also tested for antimicrobial activity. Compound 9 showed the higher antiproliferative activity for breast (MCF-7) with a TGI value of 37.67 µmol L-1, more active than positive control doxorubicin (TGI value of 46.04 µmol L-1). Compound 7 presented significant antiproliferative activity for lung, non-small cells (NCI-H460) with a TGI value of 49.52 µmol L-1 (doxorubicin, TGI value of 14.27 µmol L-1), while compound 12 showed significant activity for breast (MCF-7) with TGI value of 73.97 µmol L-1. Compound 10 showed selective activity for ovarian expressing phenotype for multiple drug resistance (NCI-ADR/RES) with a TGI value of 131.50 µmol L-1, but was less active than doxorubicin (TGI value of 14.80 µmol L-1 (Table 1). Compounds 9 and 12 also showed significant antiproliferative activity against different tumor cell lines with TGI values below to 100 µmol L-1 (Table 1).

It is important to notice that compounds 6, 7, 9, 11 and 12 shared the same basic skeleton with different substitution patterns. This way, our results suggested that a methoxylated substitute at R3 reduces antiproliferative activity (compound 11) or even results in an inactive compounds as 6, but a hydroxyl group at R3 favored the activity (compound 12). The best results were obtained for methylenedioxy group (compound 9) or methoxy groups (compounds 7 and 12) at R1 and R2.

No significant antibacterial activity was observed for the tested compounds. The only significant antimicrobial result was observed for compound 12 that showed antifungal activity against C. albicans (MIC value of 50.81 mmol L-1) similar to the positive control nystatin (MIC value of 54.00 µmol L-1).

Conclusion

The chemical investigation of the bark of G. blepharophylla has resulted in the isolation of several compounds common in the taxon Guatteria, such as, 1, 3, 6, 7 and 9 that could be considered chemotaxonomic markers of this genus. Compounds 1-6, 11 and 12 are being reported for the first time in this species, and are important for the chemotaxonomy of Annonaceae family. The significant in vitro antiproliferative results obtained against several human tumour cell lines and antimicrobial activities demonstrated by the major compounds indicate that this species is a natural source of biologically active compounds. Compound 12 showed significant antifungal and antiproliferative activities against C. albicans and human tumor cell lines of breast (MCF-7). Therefore, studies involving mechanisms of action are necessary to fully understand its biological significance.

Supplementary Information

Supplementary data including physical data for compounds 1-10 and 1H, 13C NMR, HSQC and HMBC for compounds 11 and 12 are available free of charge at http://jbcs.sbq.org.br as a PDF file.

Acknowledgements

The authors are grateful to Prof. Dr. A. C. Webber of the Universidade Federal do Amazonas (UFAM) for the botanical identification, as well as, to CAPES, CNPq and Fundação Araucária for financial support.

Submitted: July 14, 2010

Published online: February 17, 2011

FAPESP has sponsored the publication of this article

Supplementary Information

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  • 1. Erkens, R. H. J.; Maas, P. J. M.; Rodriguésia 2008, 59, 401.
  • 2. Corrêa, M. P.; Dicionário das Plantas Úteis do Brasil e das Exóticas Cultivadas, IBDF: Rio de Janeiro, Brasil, 1984.
  • 3. Silva, D. B.; Tulli, E. C. O.; Militão, G. C. G.; Costa-Lotufo, L. V.; Pessoa, C.; Moraes, M. O.; Albuquerque, S.; Siqueira, J. M.; Phytomedicine 2009, 16, 1059.
  • 4. Hsieh,T.-J.;Chang,F.-R.;Chia,Y.-C.;Chen,C.-Y.;Chiu,H.-F.; Wu, Y.-C.; J. Nat. Prod. 2001, 64, 616.
  • 5. Muhammad, I.; Dunbar, D. C.; Takamatsu, S.; Walker, L. A.; Clark, A. M.; J. Nat. Prod. 2001, 64, 559.
  • 6. Costa, E. V.; Marques, F. A.; Pinheiro, M. L. B.; Vaz, N. P.; Duarte, M. C. T.; Delarmelina, C.; Braga, R. M.; Maia, B. H. L. N. S.; J. Nat. Prod. 2009a, 72, 1516.
  • 7. Costa, E. V.; Pinheiro, M. L. B.; Silva, J. R. A.; Maia, B. H. L. N. S.; Duarte, M. C. T.; Amaral, A. C. F.; Machado, G. M. C.; Leon, L. L.; Quim. Nova 2009b, 32, 78.
  • 8. Costa, E. V.; Teixeira, S. D.; Marques, F. A.; Duarte, M. C. T.; Delarmelina, C.; Pinheiro, M. L. B.; Trigo, J. R.; Maia, B. H. L. N. S.; Phytochemistry 2008, 69, 1895.
  • 9. Costa, E. V.; Pinheiro, M. L. B.; Barison, A.; Campos, F. R.; Salvador, M. J.; Maia, B. H. L. N. S.; Cabral, E. C.; Eberlin, M. N.; J. Nat. Prod 2010, 73, 1180.
  • 10. Costa, E. V.; Pinheiro, M. L. B.; Xavier, C. M.; Silva, J. R. A.; Amaral, A. C. F.; Souza, A. D. L.; Barison, A.; Campos, F. R.; Ferreira, A. G.; Machado, G. M. C.; Leon, L. L. P. J.; J. Nat. Prod. 2006, 69, 292.
  • 11. Montenegro, H.; Gutiérrez, M.; Romero, L. I.; Ortega-Barría, E.; Capson, T. L.; Rios, L.C.; Planta Med. 2003, 69, 677.
  • 12. Mahiou, V.; Roblot, F.; Fournet, A.; Hocquemiller, R.; Phytochemistry 2000, 54, 709.
  • 13. Boyom,F.F.;Ngouana,V.;Zollo,P.H.A.;Menut,C.;Bessiere, J. M.; Gut, J.; Rosenthal, P. J.; Phytochemistry 2003, 64, 1269.
  • 14. Waechter,A.-I.;Cavé,A.;Hocquemiller;R.;Bories,C.;Muñoz, V.; Fournet, A.; Phytother. Res. 1999, 13, 175.
  • 15. Costa, E. V.; Pinheiro, M. L. B.; Marques, F. A.; Braga, R. M.; Maia, B. H. L. N. S.; Biochem. Syst. Ecol. 2009c, 37, 43.
  • 16. Maia, J. G. S.; Andrade, E. H. A.; Carreira, L. M. M.; Oliveira, J.; Araújo, J. S.; Flavour Frag. J. 2005, 20, 478.
  • 17. Shoemaker, R. H.; Nat. Rev. Cancer 2006, 6, 813.
  • 18. Ellof, J. N.; Planta Med. 1998, 64, 711.
  • 19. Ragasa,C.Y.;Ganzon,J.;Hofileña,J.;Tamboong,B.;Rideout, J. A.; Chem. Pharm. Bull. 2003, 51, 1208.
  • 20. Micheletti, A. C.; Beatriz, A.; Lima, D. P.; Honda, N. K.; Pessoa, C. O.; Moraes, M. O.; Lotufo, L. V.; Magalhães, H. I. F.; Carvalho, N. C. P.; Quim. Nova 2009, 32, 12.
  • 21. Facundo, V. A.; Polli, A. R.; Rodrigues, R. V.; Militão, J. S. L. T.; Stabelli, R. G.; Cardoso, C. T.; Acta Amaz. 2008, 38, 733.
  • 22. Guinaudeau, H.; Leboeuf, M.; Cavé, A.; J. Nat. Prod 1983, 46, 761.
  • 23. Harrigan, G. G.; Gunatilaka, A. A. L.; Kingston, D. G. I.; Chan, G. W.; Johnson, R. K.; J. Nat. Prod 1994, 57, 68.
  • 24. Arriaga, A. M. C.; Feitosa, E. M. A.; Lemos, T. L. G.; Santiago, G. M. P.; Lima, J. Q.; De Oliveira, M. C. F.; Vasconcelos, J. N. E.; Rodrigues, F. E. A.; Gomes, T. B. M.; Braz-Filho, R.; Nat. Prod. Commun. 2008, 3, 1687.
  • 25. Cortes, D.; Hocquemiller, R.; Leboeuf, M.; Cavé, A.; Morreti, C.; J. Nat. Prod 1986, 49, 878.
  • 26. Hasegawa, M.; Sojo, M.; Lira, A.; Marques, C.; Acta Cient. Venez. 1972, 23, 165.
  • 27. Fleischer, T. C.; Waigh, R. D.; Waterman, P. G.; Phytochemistry 1998, 47, 1387.
  • 28. Atti, S. A.-E.; Ammar, H. A.; Phoebe Jr., C.H.; Schiff Jr., P. L.; Slatkin, D. J.; J. Nat. Prod 1982, 45, 476.
  • *
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  • Publication Dates

    • Publication in this collection
      10 June 2011
    • Date of issue
      June 2011

    History

    • Accepted
      17 Feb 2011
    • Received
      14 July 2010
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