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Anticancer, antichemotactic and antimicrobial
activities of marine sponges collected off the coast
of Santa Catarina, southern Brazil
Noel R. Monksa,*, Clea Lernerb, Amelia T. Henriquesc,Fabiane M. Fariasc, Elfrides E.S. Schapovalc, Edna S. Suyenagac,Adriana B. da Rochaa,d, Gilberto Schwartsmanna,d, Beatriz Mothesb
aCentro Integrado do Cancer (CINCAN), Universidade Luterana do Brazil (ULBRA), Rua Miguel Tostes 101,
Canoas 92420-280, RS, BrazilbFundac�ao Zoobotanica, Museu de Ciencias Naturais, Rio Grande Sul, Rua Dr. Salvador Franca 1427,
Porto Alegre 90690-000, RS, BrazilcFaculdade de Farmacia, Universidade Federal do Rio Grande do SUL (UFRGS), Avenida Ipiranga 2752,
Porto Alegre 90610-000, RS, BrazildSouth American Office for Anticancer Drug Development (SOAD), Hospital de Clınicas de Porto Alegre,
Rua Ramiro Barcelos 2350, Porto Alegre 90035-003, RS, Brazil
Received 10 July 2002; received in revised form 22 August 2002; accepted 30 August 2002
Abstract
This study reports the in vitro screening of 10 marine sponges (Porifera) collected from the
coastline of Santa Catarina, southern Brazil, in the search for novel pharmaceuticals. Organic and
aqueous extracts were tested for anticancer, antibacterial, antifungal and antichemotactic activities.
Eight of the ten species tested demonstrated activity in one or more of the bioassays. Organic extracts
of Polymastia janeirensis Boury-Esnauls, 1973, Haliclona aff tubifera George and Wilson, 1919,
Mycale arcuiris Lerner and Hajdu, 2002 and Raspailia (syringella) sp. each demonstrated
cytotoxicity at 100 Ag/ml in an in vitro screening assay against the HT29 colorectal tumour cell line.
Further analysis against three human tumour cell lines (HT29, U373 and NCI-H460) demonstrated
IC50 concentrations ranging from 25 to 50 Ag/ml. Aqueous extracts of six species P. janeirensis, M.
arcuiris, Raspailia (syringella) sp., Guitarra sp., Tedania ignis Duchassaing and Michelotti, 1864
and Pseudaxinella reticulata Ridley and Dendy, 1886 each significantly ( pV 0.05) retarded the
migration of polymorphonuclear (PMN) leukocytes in a chemotactic assay. In antibacterial assays,
0022-0981/02/$ - see front matter D 2002 Elsevier Science B.V. All rights reserved.
PII: S0022 -0981 (02 )00380 -5
* Corresponding author. Current address: Dana-Farber Cancer Institute, Smith Building–Rm 936A, 44
Binney Street, Boston, MA 02115, USA. Tel.: +1-617-632-4172; fax: +1-617-632-4680.
E-mail address: Noel_Monks@dfci.harvard.edu (N.R. Monks).
www.elsevier.com/locate/jembe
Journal of Experimental Marine Biology and Ecology
281 (2002) 1–12
only H. aff tubifera (four of five bacterial strains) and Axinella corrugata George and Wilson, 1919
(one of five bacterial strains) demonstrated activity. None of the 10 species demonstrated measurable
antifungal activity. These extracts are currently undergoing further analysis to identify the active
constituents.
D 2002 Elsevier Science B.V. All rights reserved.
Keywords: Polymastia janeirensis; Haliclona aff tubifera; Mycale arcuiris; Raspailia (syringella) sp.
1. Introduction
Without doubt, natural products have been, and still are, the cornerstone of the health
care armamentarium. Indeed, at the last estimate, 80% of the world’s population still rely on
traditional medicines for their health care needs (Farnsworth et al., 1985). Considering
prescription medicines alone, microbial and plant-derived drugs account for greater than
30% of the worldwide sales (Grabley and Thiericke, 1999). Some of the most notable
include the analgesics aspirin (Filipendula ulmaria) and morphine and codeine (Papaver
somniferum), the malaria prophylaxis, quinine (Cinchona pubescens) and the cardiotonic
drugs, digoxin and digitoxin (Digitalis purpurea) (reviewed in Cox, 1994; da Rocha et al.,
2001).
The popularity of drug discovery programs based on nature are associated to a number of
factors. Firstly, the diversity and complexity of the chemical structures go far beyond those,
which can be synthesized in a laboratory. Secondly, the molecules isolated from nature are,
more often than not, small ( < 1000 Da), with existing drug-like properties (Harvey, 1999).
Added to this, as a result of evolutionary pressures, many organisms, both terrestrial and
marine, have developed chemical defense mechanisms, secondary metabolites, which
confer a selective advantage and often have distinct biological activities against enzymes
and receptors which makes them ideal candidates for pharmacological investigation
(Faulkner, 2000b).
In the search for new pharmaceuticals, a number of approaches are used by natural
product researchers in the selection of candidate species. These include (a) Ethnomedical
information, which refers to species used in popular medicine to treat ailments; (b)
Chemotaxonomy, which involves selection of species due to promising biological activity
or the presence of a particular class of molecule(s) with the desired activity, in congeneric
species and (c) Random collection (Farnsworth, 1994). Generally speaking, there is little or
no ethnobotanical information regarding the use of marine species for medical ailments,
therefore the screening of marine organisms generally fall into the latter category, although
once an interesting species/compound is discovered, chemotaxonomy can be used to select
related species.
Covering around 70% of the planet surface, the oceans possess a huge potential for the
new discovery often on novel molecules (Cragg et al., 1997). Compounds isolated from
marine sources are often highly complex structures, which are frequently difficult to
synthesize, which can often lead to problems in the supply of sufficient quantities of
material for preclinical and clinical development. These compounds are usually a part of
N.R. Monks et al. / J. Exp. Mar. Biol. Ecol. 281 (2002) 1–122
highly toxic defense mechanisms, which is a reflection of the highly competitive and solute
environment in which the organism resides (Grabley and Thiericke, 1999). The most
interesting phyla with respect to pharmacologically active marine compounds include
bacteria, fungi, algae, sponges, soft corals and gorgonians, sea hares and nudibranchs,
bryozoans and tunicates (Faulkner, 2000b). At present, there are a number of compounds
from marine origin which are under investigation and/or are being developed as new
pharmaceuticals (Faulkner, 2000a,b; da Rocha et al., 2001; Schwartsmann et al., 2001).
Examples include: Cytarabine (Cryptothethya crypta), Halichondrin B (Halichondria
okadai), Bryostatin 1 (Bugula neritina), Dolastatin 10 (Dolabella auricularia) and
Ecteinascidin 743 (Ecteinascidia turbinata), which are all under evaluation as new
anticancer therapies although none, as yet, are commercially available. A number of
compounds have been identified with anti-inflammatory activity, including Manoalide
(Luffariella variabilis), which is commerically available, Pseudopterosins (Pseudoptero-
gorgia elisabethae), Topsentins (Topsentia genitrix, Spongosorites sp. and Hexadella sp.),
Scytonemin (Cyanobacteria) and Debromohymenialdisine (DBH) (Phakellia flabellata,
Hymeniacidon aldis and Stylotella aurantia). Aurantosides (Siliquariaspongia japonica
and Homophymia conferta) and Spongistatin 1 (Hyrtios erecta) are commerically available
antifungal agents which were also discovered from marine sponges.
In this report, we describe the screening of marines sponge extracts collected from the
coastline of Santa Catarina, southern Brazil, for antitumour, antichemotactic, antibacterial
and antifungal activity. This study is part of a collaborative program between a number of
Brazilian institutions (Fundac�ao Zoobotanica Rio Grande do Sul, Faculdade de Farmacia-
Universidade Federal do Rio Grande do Sul and the South American Office for Anticancer
Drug Development (SOAD)) for the collection and screening of Brazilian marine sponges
for a variety of biological activities, with the aim of identifying new sponges species and
novel molecules with interesting and potentially useful therapeutic activities.
2. Materials and methods
2.1. Sponge sampling and identification
Sponges samples were collected manually from exposed and semi-exposed habitats, at
depths of between 0.5 and 14 m, from locations on the coastline of Santa Catarina
(southern Brazil). Taxonomic designation was based on scanning electron microscope
studies and on skeletal slides and dissociated spicule mounts. Specimens of all materials
are deposited in the Museu de Ciencias Naturais–Porifera (MCNPOR) collection of the
Fundac�ao Zoobotanica do Rio Grande do Sul, Brazil. The species investigated in this studyare detailed in Table 1.
2.2. Extract preparation
Aqueous extracts were produced by the following procedure. Sponge materials were
ground together with sand and water three times for 30 min. The resulting extract (collected
after each 30 min) was subsequently filtered and freeze-dried. The remaining material was
N.R. Monks et al. / J. Exp. Mar. Biol. Ecol. 281 (2002) 1–12 3
sequentially extracted five times with a methanol/toluene mixture (3:1, v/v) by maceration
over 5 days. The resulting extract solution was then filtered and concentrated in a
Rotavapor. For the chemotatic, antibacterial and antifungal assays, both the aqueous and
organic extracts were suspended in Hanks buffer and Tween 80 (9:1, v/v) at a concentration
of 100 Ag/ml (w/v). The preparation of the extracts for the in vitro antitumour assays is
described in Section 2.4.
2.3. Cell culture maintenance
The HT29 human colon adenocarcinoma (ATCC No. HTB-38), NCI-H460 human
large cell lung carcinoma (ATCC No. HTB-177) and U373 human glioblastoma astrocy-
toma (ECACC No. 89081403) cell lines were maintained as exponentially growing
cultures in RPMI 1640 culture medium, supplemented with 10% foetal bovine serum,
pH 7.4. All cell lines were cultured at 37 jC in an atmosphere of 5% CO2 in air (100%
humidity).
2.4. Cytotoxic screening assay
As part of our general anticancer screening programme, all extracts were initially tested
at a concentration of 100 Ag/ml against HT29 cells to eliminate extracts which did not
demonstrate cytotoxic activity. HT29 cells were seeded into 96-well cell culture plates and
incubated overnight to allow adherence. The extracts were dissolved in 100% DMSO and
added to the wells in triplicate at a final concentration of 100 Ag/ml (final DMSO
concentration 0.25% (v/v) at which no growth inhibitory effects were observed). Both
culture medium alone and culture medium plus vehicle (0.25% DMSO) controls were
used. Following addition of the extracts, plates were incubated for 72 h, after which
cellular growth was determined using the sulforhodamine B (SRB) colourimetric protein
assay (Skehan et al., 1990). Extracts which produced an SRB absorbance lower than that
of the time-zero value (approximately 10% of the control growth), generated by cellular
fixation, using 50% TCA, immediately prior to the addition of the extracts, were
considered to be cytotoxic and submitted for further investigation.
Table 1
Sponge species examined in this study
Species Family Authors Extracts testeda
Guitarra sp. Guitarridae O+A
P. citrina Halichondriidae Muricy et al., 2001 O+A
T. ignis Tedaniidae Duchassaing and Michelotti, 1864 O+A
P. reticulata Axinellidae Ridley and Dendy, 1886 O+A
P. janeirensis Polymastiidae Boury-Esnauls, 1973 O+A
A. corrugata Axinellidae George and Wilson, 1919 O+A
H. aff tubifera Chalinidae George and Wilson, 1919 O+A
Guitarra sp. Guitarridae O+A
M. arcuiris Mycalidae Lerner and Hajdu, 2002 O+A
Raspailia (Syringella) sp. Raspailiidae O+A
a O—Organic extract, A—Aqueous extract.
N.R. Monks et al. / J. Exp. Mar. Biol. Ecol. 281 (2002) 1–124
2.5. Growth inhibition assay
The IC50 (concentration at which cellular growth is inhibited by 50%) was determined
against HT29, NCI-H460 and U373 cells (3.5, 1 and 2.5� 103 cells/well, respectively)
using the methods described above (Section 2.4). Cells were treated in triplicate with a
log10 concentration range (0.1, 1, 10 and 100 Ag/ml) of each extract for 72 h. The IC50
values were estimated from a semi-log plot of extract concentration against SRB
absorbance as a percentage of vehicle control growth (0.25% DMSO).
2.6. Antichemotactic assay
Chemotaxis was measured by the method described previously by Zigmond and Hirsch
(1973). Prior to the chemotactic assay, rat leukocytes were treated with 100 Ag/ml of
sponge suspension at 37 jC for 1 h. Plasma collected from rats was incubated at 37 jC for
30 min with 65 Ag/ml of lipopolysaccharide (from Escherichia coli), following which the
plasma was diluted in Hanks buffer 1:5 (v/v). Chemotaxic migration of leukocytes through
an 8.0-Am nitrocellulose filter, towards the chemotactic stimulant (lipopolysaccharide-
treated plasma), was measured after incubation for 1 h at 37 jC using the micrometer on
the fine-focus knob of the microscope. The distance from the top of the filter to the farthest
plane of focus still containing two cells, in five microscopic fields, allowed the evaluation
of leukocyte migration. The assay was carried out in duplicate.
2.7. Antibacterial assay
Antibacterial activity was determined against cultures of E. coli (ATCC 25922),
Staphylococcus aureus (ATCC 6538P), Staphylococcus epidermidis (ATCC 12228),
Bacillus subtilis (ATCC 6633) and Micrococcus luteus (ATCC 9341) using the agar-
diffusion assay method (Limberger et al., 2001). Ten plates were tested against each sponge
sample at 2.5 mg/ml (five per aqueous extract and five per organic extract). Chloramphe-
nicol (400 Ag/ml) was used as a positive control. Following incubation at 37 jC for 24 h, the
diameters (mm) of the growth inhibition halos were determined using a pachymeter.
2.8. Antifungal assay
Cultures of Candida albicans (ATCC 10231) and Saccharomyces cerevisiae (ATCC
1600) were treated with sponge extracts (2.5 mg/ml) as previously described in the
antibacterial assay (see Section 2.7). Nistatin was used as a positive control at a
concentration of 3 mg/ml. Following incubation at 25 jC for 18 h, the diameters (mm)
of the growth inhibition halos were measured using a pachymeter.
3. Results and discussion
Table 2 shows the results of the in vitro testing of sponge extracts against the HT29
colorectal tumour cell line. This is the first step in our anticancer drug development
N.R. Monks et al. / J. Exp. Mar. Biol. Ecol. 281 (2002) 1–12 5
programme and is designed to identify those extracts with cytotoxic activity. Four sponge
species (organic extracts) were found to be cytotoxic at 100 Ag/ml, namely, Polymastia
janeirensis, Haliclona aff tubifera, Mycale arcuiris and Raspailia (syringella) sp. These
four active extracts were further tested against three human tumour cell lines, derived from
different tumour types, over a concentration range (0.1–100 Ag/ml) to determine their
potency (IC50–50% inhibition of cell growth), the results of these tests are shown in Table
3. The four extracts displayed similar levels of activity across the panel of three cell lines,
the IC50 values ranged from 25 to 50 Ag/ml. These extracts are currently undergoing
further investigation through collaboration with the Natural Products Branch of the
American National Cancer Institute (NCI). Using a 60-cell line panel derived from a
range of different tumour types, the NCI can help identify extracts with interesting
activities by the pattern and degree of activity across their human tumour cell line panel.
To date, a number of growth inhibitory, cytotoxic and other pharmacologically active
molecules have been isolated from species of Porifera related to those tested in this study;
these include alkaloids, macrolides and peptides and are summarized in Table 4.
Table 2
In vitro screening of sponge extracts (100 Ag/ml) against the HT29 human colorectal tumour cell line
Species Cell growtha (% of control growth)
Organic Aqueous
Guitarra sp. 92b 89F 13
P. citrina 68F 19 97F 3
T. ignis 44F 9 90F 10
P. reticulata 91F 8 94F 4c
P. janeirensis 0.4F 0.3* 65F 17
A. corrugata na 94F 2
H. aff tubifera 1F1* 37F 5
Guitarra sp. 44F 5 103F 3
M. arcuiris 2F 1* 63F 17
Raspailia (Syringella) sp. 1F1* 104F 6
a Values were determined using the SRB protein assay after 72 h continuous exposure to the extracts. All
values given are the meanF S.D. of z 3 separate experiments, unless otherwise stated. na =Not tested.b n= one experiment.c n= two experiments.
*Extracts with cytotoxic activity at 100 Ag/ml (i.e. those extracts whose growth is less than the time-zero
control = 10% of control growth).
Table 3
In vitro growth inhibitory activity of sponge extracts against human tumour cell lines
Species Extract type IC50 (Ag/ml)a
HT29 U373 NCI-H460
H. aff tubifera Organic 28F 4 31F 3 30F 1
P. janeirensis Organic 42F 7 33F 3 28F 5
M. arcuiris Organic 31F1 50F 17 27F 1
Raspailia (Syringella) sp. Organic 27F 6 32F 2 25F 0
a IC50 values were determined using the SRB protein assay after 72 h continuous exposure to the extracts. All
values given are the meanF S.D. of z 3 separate experiments.
N.R. Monks et al. / J. Exp. Mar. Biol. Ecol. 281 (2002) 1–126
The inflammatory reaction consists of three fundamental processes: (1) hemodynamic
changes, (2) alterations in vessel permeability and (3) accumulation of inflammatory cells.
The directed migration of inflammatory cells along a chemical gradient is termed che-
Table 4
Literature reports of cellular activity
Species Compounds Chemical class Pharmacological
activity
Reference
Axinella sp. Halichondrin B,
Homohalichondrin B
Macrocyclic
lactones
Growth inhibitory Pettit et al., 1994a
Axinastatins Cycloheptapeptides Growth inhibitory Pettit et al., 1994a,b
Hymenistatin Cyclo-octapeptide Growth inhibitory Pettit et al., 1994b
A. carteri Hymenialdisines Alkaloids Cytotoxicity Supriyono et al., 1995
Halistatin 2 Macrocyclic
lactones
Binds tubulin Luduena et al., 1995
A. weltneri Sodwanones Triterpenes Cytotoxicity Rudi et al., 1997, 1999
Haliclona sp. Haliclonacyclamines Alkaloids Cytotoxicity Charan et al., 1996;
Clark et al., 1998
Adociasulfates Meroterpenoid Inhibits Kinesin Blackburn and Faulkner,
2000; Blackburn
et al., 1999
Haliclamide Cyclic depsipeptide Cytotoxicity Randazzo et al., 2001
Salicylihalamides Macrolides Cytotoxicity Erickson et al., 1997
Manzamines Alkaloids Cytotoxicity Shen et al., 1996
Keramaphidin B Alkaloid Cytotoxicity Shen et al., 1996
Haliclamines Alkaloids Cytotoxicity Fusetani et al., 1989
H. viridis HvTX toxin Nerve potassium
permeability
Sevcik et al., 1994
Halitoxin Alkylpyridine In vivo antitumour Baslow and
Turapaty, 1969
H. negra Haligramides Hexapeptides Cytotoxicity Rashid et al., 2000
H. exigua Araguspongin C,
Xestospongin D
Inhibitor of nitric
oxide synthetase
Venkateswara Rao
et al., 1998
H. tulearensis Halitulin Alkaloid Cytotoxicity Kashman et al., 1999
H. osiris Osirisynes Oygenated
polyacetylenes
Cytotoxicity,
inhibitors
of Na+/K+-ATPase.
Shin et al., 1998
Mycale sp. Mycalamides Mycalamides Cytotoxicity Ogawara et al., 1991;
Perry et al., 1988, 1990;
West et al., 2000b
Pateamine Macrolide Cytotoxicity,
Immunosupressant
Hood et al., 2001;
Remuinan and
Pattenden, 2000
Peloruside A Macrolide Cytotoxicity West et al., 2000a
Thiomycalolides Macrolides Cytotoxicity Matsunaga et al., 1998a
Onnamide Cytotoxicity Burres and Clement, 1989
Mycalisines
A and B
Nucleosides Inhibitors of
cell division
Kato et al., 1985
M. magellanica Mycalolides Macrolides Cytotoxicity Matsunaga et al., 1998b
M. micracanthoxea Mycalazols Pyrroles Cytotoxicity Ortega et al., 1997
Raspailia sp. Asmarines Terpenoids Cytotoxicity Yosief et al., 2000, 1998
N.R. Monks et al. / J. Exp. Mar. Biol. Ecol. 281 (2002) 1–12 7
motaxis. The activation of this process appears to be an important mechanism by which
the immune effector cells are located at sites of inflammation. Based on this, extracts from
marine sponges were analyzed for their activity on polymorphonuclear (PMN) leukocytes
chemotaxis. Six species, P. janeirensis, M. arcuiris, Raspailia (syringella) sp., Guitarra
sp., Tedania ignis and Pseudaxinella reticulata all significantly ( pV 0.05) reduced the
migration of the leukocytes through a nitrocellulose filter towards the chemotatic
stimulant (Table 5). Interestingly, the antichemotactic activity was seen solely in the
aqueous extracts, whereas extracts which demonstrated in vitro cytotoxicity were all from
the organic phase.
Two of the sponge species tested demonstrated antibacterial activity (Table 6). The
aqueous extract from Axinella corrugata showed weak activity (7–11 mm halo) againstM.
luteus. The organic extract from H. aff tubifera demonstrated activity against each of the
bacteria tested (weak, 7–11 mm), with the exception of B. subtilis, the strongest activity
(moderate, 11–16 mm halo) was seen against E. coli. The aqueous extract from H. aff
tubifera showed weak activity againstM. luteus. A number of antimicrobial molecules have
already been isolated from related sponge species, these are outlined in Table 7.
None of the species tested demonstrated activity in antifungal tests against the species C.
albicans and S. cerevisiae (Nistatin—C. albicans and S. cerevisiae, 7–11 and 11–16 mm
halos, respectively).
Table 5
Antichemotatic activity of marine sponge extracts
Sponge species Extracta PMN migrationb(Am)
Control 123.0F 0.1
Guitarra sp. O 122.7F 2.1
A 40.0F 0.1*
P. citrina O 121.0F 1.1
A 121.2F 2.0
T. ignis O 122.3F 1.5
A 18.9F 0.0*
P. reticulata O 121.3F 1.1
A 25.3F 0.0*
P. janeirensis O 122.3F 1.5
A 19.0F 0.02*
A. corrugata O C
A Cy
H. aff tubifera O C
A 122.17F 2.19
Guitarra sp. O Cy
A 119.0F 3.0
M. arcuiris O 122.67F 1.3
A 13.0F 0.02*
Raspailia (syringella) sp. O Cy
A 35.33F 1.03*
All values given are the meanF S.D. of 10 separate experiments.a A—Aqueous extract, O—Organic extract.b C—Cellular wall damage, Cy—Cytotoxic.
*Statistically significant—p< 0.05 (Student’s t-test).
N.R. Monks et al. / J. Exp. Mar. Biol. Ecol. 281 (2002) 1–128
Over the past decade, interest in marine natural products has dramatically increased, and
consequently, a number of novel molecules have, or are being, developed as pharmaceut-
icals, e.g. Cytarabine, Ecteinascidin 743, Manoalide and Spongistatin 1 to name only a few.
Table 6
Antibacterial activity of the Brazilian marine sponges
Species Extract E. coli S. aureus S. epidermidis B. subtilis M. luteus
Chloroamphenicol *** ** *** ** ***
Guitarra sp. O – – – – –
A – – – – –
P. citrina O – – – – –
A – – – – –
T. ignis O – – – – –
A – – – – –
P. reticulata O – – – – –
A – – – – –
P. janeirensis O – – – – –
A – – – – –
A. corrugata O – – – – –
A – – – – *
H. aff tubifera O ** * * – *
A – – – – *
Guitarra sp. O – – – – –
A – – – – –
M. arcuiris O – – – – –
A – – – – –
Raspailia (Syringella) sp. O – – – – –
A – – – – –
(– ) No activity, (*) weak activity (7–11 mm halo), (**)moderate activity (11–16 mm halo), (***) high activity
(>16 mm halo).
Table 7
Literature reports of antimicrobial activity
Species Compounds Chemical
class
Pharmacological
activity
Reference
Axinella sp. Axinellamines Alkaloids Antibacterial Urban et al., 1999
A. polycapella Hydroxyhydroquinone,
2,2V,4,4V,5,5V-Hexahydro-biphenylAntimicrobial Wratten and Meinwald,
1981
Haliclona sp. Crude extract Antifungal Bhosale et al., 1999
Haliclonacyclamines Alkaloids Antibacterial,
Antifungal.
Charan et al., 1996
Haliclotriol Terpene-
ketides
Antibacterial Crews and Harrison,
2000
Haliclonadiamine Alkaloid Antimicrobial Fahy et al., 1988
Papuamine Alkaloid Antifungal Baker et al., 1988
H. osiris Osirisynes Oygenated
polyacetylenes
Inhibitors of
reverse
transcriptase
Shin et al., 1998
Mycale sp. Mycalamides Mycalamides Antiviral Perry et al., 1988,
1990
N.R. Monks et al. / J. Exp. Mar. Biol. Ecol. 281 (2002) 1–12 9
Often present as defense mechanisms, these potent marine compounds provide pharma-
ceutical researchers with a novel platform for the development of new drugs to treat serious
diseases such as cancer, bacterial infection and arthritis. Here, we have shown that a number
of sponge species have activity in in vitro models systems, which are directly relevant to
human disease. Studies of this nature highlight the potential of marine product screening
programmes through which, without question, will identify new drugs from the vast,
untapped, resources that are within our oceans.
4. Conclusions
Sponges species collected from the State of Santa Catarina in southern Brazil have been
shown to posses a number of biological activities. The most interesting species are H. aff
tubifera, P. janeirensis, M. arcuiris and Raspailia (syringella) sp.—antineoplastic; P.
janeirensis, M. arcuiris, Raspailia (syringella) sp., Guitarra sp., T. ignis and P. reticu-
lata—antichemotactic and A. corrugata and H. aff tubifera—antibacterial. To the best of
our knowledge, this is the first report demonstrating antineoplastic, antichemotactic and
antibacterial activity of these species of porifera. These species are currently undergoing
detailed investigations with the objective of isolating and identifying the molecular species
responsible for the activities demonstrated in this report. Furthermore, the encouraging
biological activities seen in this study show that the Brazilian coastline is a potential
source of sponge species worthy of further investigation. In light of the work presented
here, we have initiated further collection programs off the coastlines of Sao Paulo
(southeast) and Pernambuco (northeast).
Acknowledgements
This work was supported by grants from FAPERGS, CNPq, CAPES and the SOAD
foundation. The authors would also like to thank Dr. Miriam Anders Apel for her technical
assistance with this work. [SS]
References
Baker, B.J., Scheuer, P.J., Shoorlery, J.N., 1988. Papuamine: an antifungal pentacyclic alkaloid from a marine
sponge. J. Am. Chem. Soc. 110, 965–966.
Baslow, M.H., Turapaty, P., 1969. In vivo antitumor activity and other pharmacological properties of halitoxin
obtained from the sponge Haliclona viridis. Proc. West. Pharmacol. Soc. 12, 6–8.
Bhosale, S.H., Jagtap, T.G., Naik, C.G., 1999. Antifungal activity of some marine organisms from India against
food spoilage Aspergillus strains. Mycopathologia 147, 133–138.
Blackburn, C.L., Faulkner, D.J., 2000. Adociasulfate 10: a new merohexaprenoid sulfate from the sponge
Haliclona (aka Adocia) sp. Tetrahedron 56, 8429–8432.
Blackburn, C.L., Hopmann, C., Sakowicz, R., Berdelis, M.S., Goldstein, L.S.B., Faulkner, D.J., 1999. Adocia-
sulfates 1–6: inhibitors of kinesin motor proteins from the Halicona (aka Adocia) sp. J. Org. Chem. 64,
5565–5570.
Burres, N.S., Clement, J.J., 1989. Antitumor activity and mechanism of action of the novel marine natural
products mycalamide-A and -B, and onnamide. Cancer Res. 49, 2935–2940.
N.R. Monks et al. / J. Exp. Mar. Biol. Ecol. 281 (2002) 1–1210
Charan, R.D., Garson, M.J., Brereton, I.M., Willis, A.C., Hooper, J.N.A., 1996. Haliclonacylamines A and B:
cytotoxic alkaloids from the tropical marine sponge Haliclona sp. Tetrahedron 52, 9111–9120.
Clark, R.J., Field, K.L., Charan, R.D., Garson, M.J., Brereton, I.M., Willis, A.C., 1998. The haliclonacycla-
mines: cytotoxic tertiary alkaloids from the tropical marine sponge Haliclona sp. Tetrahedron 54,
8811–8826.
Cox, P.A., 1994. The ethnobotanical approach to drug discovery: strengths and limitations. In: Chadwick, D.J.,
Marsh, J. (Eds.), Ethnobotany and the Search for New Drugs. Ciba Foundation Symposium, vol. 185. Wiley,
Chichester, pp. 25–41.
Cragg, G.M., Newman, D.J., Weiss, R.B., 1997. Coral reefs, forests, and thermal vents: the worldwide explora-
tion of nature for novel antitumor agents. Semin. Oncol. 24, 156–163.
Crews, P., Harrison, B., 2000. New triterpene-ketides (Merotriterpenes), haliclotriol A and B, from an Indo-
Pacific Haliclona sponge. Tetrahedron 56, 9039–9046.
da Rocha, A.B., Lopes, R.M., Schwartsmann, G., 2001. Natural products in anticancer therapy. Curr. Opin.
Pharmacol. 1, 364–369.
Erickson, K.L., Beutler, J.A., Cardellina II, J.H., Boyd, M.R. 1997. Salicylhalamides A and B: novel cytotoxic
macrolides from the larine sponge Haliclona sp. J. Org. Chem. 62, 8188–8192.
Fahy, E., Molinski, T., Harper, M.K., Sullivan, B.W., Faulkner, D.J., Parkanyi, L., Clardy, J., 1988. Haliclonadia-
mine: an antimicrobial alkaloid from the sponge Haliclona sp. Tetrahedron Lett. 29, 3427–3428.
Farnsworth, N.R., 1994. Ethnopharmacology and drug development. In: Chadwick, D.J., Marsh, J. (Eds.),
Ethnobotany and the Search for New Drugs. Ciba Foundation Symposium, vol. 185. Wiley, Chichester,
pp. 42–59.
Farnsworth, N.R., Akerele, O., Bingel, A.S., Soejarto, D.D., Guo, Z., 1985. Medicinal plants in therapy. B. World
Health Organ. 63, 965–981.
Faulkner, D.J., 2000a. Highlights of marine natural products chemistry (1972–1999). Nat. Prod. Rep. 17, 1–6.
Faulkner, D.J., 2000b. Marine pharmacology. Antonie Van Leeuwenhoek 77, 135–145.
Fusetani, N., Yasumuro, K., Matsunaga, S., Hirota, H., 1989. Haliclamines A and B: cytotoxic macrocyclic
alkaloids from a sponge of the genus Haliclona. Tetrahedron Lett. 30, 6891–6894.
Grabley, S., Thiericke, R., 1999. Bioactive agents and natural sources: trends in discovery and application. Adv.
Biochem. Eng. Biotechnol. 64, 101–154.
Harvey, A.L., 1999. Medicines from nature: are natural products still relevant to drug discovery? Trends Phar-
macol. Sci. 20, 196–198.
Hood, K.A., West, L.M., Northcote, P.T., Berridge, M.V., Miller, J.H., 2001. Induction of apoptosis by the marine
sponge (Mycale) metabolites, mycalamide A and pateamine. Apoptosis 6, 207–219.
Kashman, Y., Koren-Goldshlager, G., Gravalos, M.D.G., Schleyer, M., 1999. Halitulin: a new cytotoxic alkaloid
from the marine sponge Haliclona tulearensis. Tetrahedron Lett. 40, 997–1000.
Kato, Y., Fusetani, N., Matsunaga, S., Hashimoto, K., 1985. Bioactive metabolites: IX. Mycalisines A and B:
novel nucleosides which inhibit cell division of fertilised starfish eggs from the marine sponge Mycale sp.
Tetrahedron Lett. 26, 3483–3486.
Limberger, R.P., Sobral, M.E.G., Zuanazzi, J.A.S., Moreno, P.R.H., Schapoval, E.E.S., Henriques, A.T., 2001.
Biological activities and essential oil composition of leaves of Blepharoclyx salicifolius. Pharm. Biol. 39,
308–311.
Luduena, R.F., Roach, M.C., Prasad, V., Pettit, G.R., Cochacz, Z.A., Herald, C.L., 1995. Interaction of three
sponge-derived macrocyclic lactone polyethers (spongistatin 3, halistatins 1 and 2) with tubulin. Drug Dev.
Res. 35, 40–48.
Matsunaga, S., Nogata, Y., Fusetani, N., 1998a. Thiomycaloides: new cytotoxic trisoxazole-containing macro-
lides isolated from a marine sponge Mycale sp. J. Nat. Prod. 61, 663–666.
Matsunaga, S., Sugawara, T., Fusetani, N., 1998b. New mycaloides from the marine sponge Mycale magellanica
and their interconversion. J. Nat. Prod. 61, 1164–1167.
Ogawara, H., Higashi, K., Uchino, K., Perry, N.B., 1991. Change of Ras-transformed NRK-cells back to normal
morphology by mycalamides A and B: antitumor agents from a marine sponge. Chem. Pharm. Bull. 39,
2152–2154.
Ortega, M.J., Zubia, E., Carballo, J.L., Salva, J., 1997. New cytotoxic matabolites from the marine sponge
Mycale micracanthoxea. Tetrahedron 53, 331–340.
N.R. Monks et al. / J. Exp. Mar. Biol. Ecol. 281 (2002) 1–12 11
Perry, N.B., Blunt, J.W., Munro, M.H.G., 1988. Mycalamide A: an antiviral compound from a New Zealand
sponge of the genus Mycale. J. Am. Chem. Soc. 110, 4850–4851.
Perry, N.B., Blunt, J.W., Munro, M.H.G., Thompson, A.M., 1990. Antiviral and antitumor agents from a New
Zealand sponge,Mycale sp.: 2. Structures and solution conformations of mycalamides A and B. J. Org. Chem.
55, 223–227.
Pettit, G.R., Gao, F., Cerny, R.L., Doubek, D.L., Tackett, L.P., Schmidt, J.M., Chapuis, J.C., 1994a. Antineo-
plastic agents 278. Isolation and structure of axinastatins 2 and 3 from a western Caroline Island marine
sponge. J. Med. Chem. 37, 1165–1168.
Pettit, G.R., Gao, F., Schmidt, J.M., Chapuis, J.C., Cerny, R.L., 1994b. Isolation and structure of axinastatin 5
from a Republic of Comoros marine sponge. Bioorg. Med. Chem. Lett. 4, 2935–2940.
Randazzo, A., Debitus, C., Gomez-Paloma, L., 2001. Haliclamide: a novel cyclic metabolite from the Vanuatu
marine sponge Haliclona sp. Tetrahedron 57, 4443–4446.
Rashid, M.A., Gustafson, K.R., Boswell, J.L., Boyd, M.R., 2000. Haligramides A and B: two new cytotoxic
hexapeptides from the marine sponge Haliclona nigra. J. Nat. Prod. 63, 956–959.
Remuinan, M.J., Pattenden, G., 2000. Total synthesis od (� )-pateamine: a novel polyene bis-macrolide with
immunosuppressive activity from the sponge Mycale sp. Tetrahedron Lett. 41, 7367–7371.
Rudi, A., Aknin, M., Gaydou, E.M., Kashman, Y., 1997. Sodwanones K, L and M: new triterpenes from the
marine sponge Axinella weltneri. J. Nat. Prod. 60, 700–703.
Rudi, A., Yosief, T., Schleyer, M., Kashman, Y., 1999. Several new isprenoids from two marine sponges of the
family Axinellidae. Tetrahedron 55, 5555–5566.
Schwartsmann, G., da Rocha, A.B., Berlinck, J.G.S., Jimeno, J., 2001. Marine organisms as a source of new
anticancer agents. Lancet Oncol. 2, 221–225.
Sevcik, C., Garcia-Rodriguez, A.I., D’Suze, G., Mijares, A.J., 1994. Specific blockage of squid axon resting
potassium permeability by Haliclona viridis (Porifera: Haliclonidae) toxin (HvTX). Toxicon 32, 773–788.
Shen, Y.C., Tai, H.R., Duh, C.Y., 1996. Bioactive constituents from Haliclona sp.: a Formosan marine sponge.
Chin. Pharmacol. J. 48, 1–10.
Shin, J., Seo, Y., Cho, K.W., Rho, J.R., Paul, V.J., 1998. Osirisynes A–F: highly oxygenated polyacetylenes from
the sponge Haliclona osiris. Tetrahedron 54, 8711–8720.
Skehan, P., Storeng, R., Scudiero, D., Monks, A., McMahon, J., Vistica, D., Warren, J.T., Bokesch, H., Kenney,
S., Boyd, M.R., 1990. New colorimetric cytotoxicity assay for anticancer-drug screening. J. Natl. Cancer Inst.
82, 1107–1112.
Supriyono, A., Schwarz, B., Wray, V., Witte, L., Muller, W.E., van Soest, R., Sumaryono, W., Proksch, P., 1995.
Bioactive alkaloids from the tropical marine sponge Axinella carteri. Z. Naturforsch. 50, 669–674.
Urban, S., De Almeida Leone, P., Carroll, A.R., Fechner, G.A., Smith, J., Hooper, J.N.A., Quinn, R.J., 1999.
Axinellamines A–D: novel imidazo-azolo-imidazole alkaloids from the Australian marine sponge Axinella
sp. J. Org. Chem. 64, 731–735.
Venkateswara Rao, J., Desaiah, D., Vig, P.J., Venkateswarlu, Y., 1998. Marine biomolecules inhibit rat brain nitric
oxide synthetase activity. Toxicology 129, 103–112.
West, L.M., Northcote, P.T., Battershill, C.N., 2000a. Peloruside A: a potent cytotoxic macrolide isolated from the
New Zealand marine sponge Mycale sp. J. Org. Chem. 65, 445–449.
West, L.M., Northcote, P.T., Hood, K.A., Miller, J.H., Page, M.J., 2000b. Mycalamide D: a new cytotoxic amide
from the New Zealand marine sponge Mycale species. J. Nat. Prod. 63, 707–709.
Wratten, S.J., Meinwald, J., 1981. Antimicrobial metabolites of the marine sponge Axinella polycapella. Expe-
rientia 37, 13–14.
Yosief, T., Rudi, A., Stein, Z., Goldberg, I., Gravalos, G.M.D., Schleyer, M., Kashman, Y., 1998. Asmarines A–C:
three novel cytotoxic metabolites from the marine sponge Raspailia species. Tetrahedron Lett. 39, 3323–3326.
Yosief, T., Rudi, A., Kashman, Y., 2000. Asmarines A–F: novel cytotoxic compounds from the marine sponge
Raspailia sp. J. Nat. Prod. 63, 299–304.
Zigmond, S.H., Hirsch, J.G., 1973. Leukocyte locomotion and chemotaxis. New methods for the evaluation and
demonstration of a cell-derived chemotatic factor. J. Exp. Med. 137, 387–410.
N.R. Monks et al. / J. Exp. Mar. Biol. Ecol. 281 (2002) 1–1212
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