out_35.pdf

REVIEW PAPER

Cyanobacteria: potential candidates for drug discovery

Rakhi Bajpai Dixit M. R. Suseela

Received: 7 December 2012 / Accepted: 28 February 2013 / Published online: 27 March 2013

Springer Science+Business Media Dordrecht 2013

Abstract Cyanobacteria are a rich source of vast

array of bioactive molecules including toxins with

wide pharmaceutical importance. They show varied

bioactivities like antitumor, antiviral, antibacterial,

antifungal, antimalarial, antimycotics, antiprolifera-

tive, cytotoxicity, immunosuppressive agents and

multi-drug resistance reversers. A number of tech-

niques are now developed and standardized for the

extraction, isolation, detection and purification of

cyanobacterial bioactive molecules. Some of the

compounds are showing interesting results and have

successfully reached to phase II and phase III of

clinical trials. These compounds also serve as lead

compounds for the development of synthetic ana-

logues with improved bioactivity. Cyanobacterial

bioactive molecules hold a bright and promising

future in scientific research and great opportunity for

drug discovery. This review mainly focuses on

anticancerous, antiviral and antibacterial compounds

from cyanobacteria; their clinical status; extraction

and detection techniques.

Keywords Bioactive molecules Anticancerous Antiviral Antimicrobial Clinical trials Extraction methods

Introduction

Cyanobacteria are a group of photosynthetic prokary-

otes and are among the most successful and oldest life

forms present on earth (Gademann and Portmann

2008; Bajpai et al. 2010). They represent an excep-

tionally diverse but highly specialized group of micro-

organisms adapted to various ecological habitats.

They can be found in terrestrial, glaciers, aerial,

marine, brackish and fresh water environments. Cya-

nobacteria are often a main component of phytoplank-

ton in many freshwater and marine ecosystems.

Cyanobacteria produce one or a range of bioactive

compounds, which are potentially rich source of a vast

array of products with applications in feed, food,

nutritional, cosmetic, pharmaceutical and neutraceu-

tical industries (Tan 2007). Due to their high chemical

stability and water solubility, these compounds have

important implications. They have a bright future in

scientific research and for human welfare.

According to World Health Organisation (WHO),

approximately 80 % of the world population depends

on traditional remedies for their primary health care

needs. Since, thousands of years natural products have

been found to be used for treating various diseases and

they form a major milestone for modern therapeutics.

In the microbial world, especially cyanobacteria are

prolific producers of secondary metabolites, many of

which show various biological activities or bioactiv-

ity. Gerwick et al. (2008) found that secondary

metabolites are mostly isolated from the members of

R. B. Dixit (&) M. R. SuseelaAlgology Section, CSIR-National Botanical Research

Institute, Lucknow 226001, Uttar Pradesh, India

e-mail: [email protected]

123

Antonie van Leeuwenhoek (2013) 103:947961

DOI 10.1007/s10482-013-9898-0

oscillatoriales (49 %), followed by nostocales (26 %),

chroococcales (16 %), pleurocapsales (6 %) and

stigonematales (4 %). Cyanobacteria such as Ana-

baena, Nostoc, Microcystis, Lyngbya, Oscillatoria,

Phormidium and Spirulina produce variety of high

value compounds such as carotenoids, fatty acids,

lipopeptides, polysaccharides and other bioactive

compounds. Apoptogenic activity was more abundant

in the genera Anabaena and Microcystis compared to

Nostoc, Phormidium, Planktothrix, and Pseudoanaba-

ena (Oftedal et al. 2011). Interestingly synthesis of

these biomolecules remains an enigma and unresolved

puzzle to the scientific world.

A majority of these biomolecules are peptides and

are synthesized by large multimodular nonribosomal

polypeptide (NRPS) or mixed polyketide (PKS)-

NRPS enzymatic systems (Schwarzer et al. 2003). In

aquatic environments, these metabolites usually

remain within the microbial cells and are released in

substantial amounts on cell lysis (Chorus and Bartram

1999). Richard E. Moore (1970s to early 2000s),

revealed that the marine cyanobacteria are an excep-

tionally rich source of secondary metabolites (Car-

dellina and Moore 2010). Cyanobacterial secondary

metabolites includes different compounds like cyto-

toxic (41 %), antitumor (13 %), antiviral (4 %),

antimicrobial (12 %) and other compounds (18 %)

include antimalarial, antimycotics, multi-drug resis-

tance reversers, antifeedant, herbicides and immuno-

suppressive agents (Burja et al. 2001). Thus,

cyanobacteria continue to be explored and their

metabolites are now evaluated in number of biological

areas and they are becoming an exceptional source of

leading compounds for drug discovery (Singh et al.

2005, 2011; Nunnery et al. 2010; Bajpai et al. 2010).

Anticancerous, antiviral and antibacterial bioactive

molecules produced by various cyanobacteria are

listed in Table 1. These natural products not only serve

directly as drugs, but also being used as template for

the discovery and synthesis of new drugs.

Anticancerous compounds

A large number of cyanobacterial bioactive com-

pounds are found to target tubulin or actin filaments in

eukaryotic cells, making them an attractive source of

anticancer agents (Jordan and Wilson 1998). The

small anticancerous peptide, Dolastatin 10 and

Dolastatin 12 was isolated from Symploca sp. and

Leptolyngbya sp. (Kalemkerian et al. 1999; Catassi

et al. 2006). Curacin A shows antiproliferative activity

that has been isolated from cyanobacterium Lyngbya

majuscula (Nagle et al. 1995). It is also artificially

synthesized because of its pharmacological impor-

tance (Muir et al. 2002). Cryptophycin isolated from

Nostoc shows dose-dependent inhibition of L1210

leukemia cell line (Smith et al. 1994). Rickards et al.

(1999) isolated two compounds calothrixin A and B

from the organic extracts of Calothrix strains and

found that it inhibited the growth of human HeLa

cancer cells.

Apratoxin A from L. majuscula (Luesch et al.

2001a), Apratoxin BC from Lyngbya sp. (Luesch

et al. 2002a), Apratoxin D from L. majuscula and

Lyngbya sordid (Gutierrez et al. 2008), Apratoxin E

from Lyngbya bouilloni (Matthew et al. 2008) and

Apratoxins FG from L. bouilloni (Tidgewell et al.

2010) showed cytotoxicity to various cancer cell lines

i.e. U2OS osteosarcoma, HT29 colon adenocarci-

noma, HeLa cervical carcinoma, KB oral epidermoid

cancer, LoVo colon cancer, H-460 lung cancer and

HCT-116 colorectal cancer cells lines. Luesch et al.

(2000) isolated Lyngbyabellin B from L. majuscula, it

shows cytotoxic against KB and LoVo cells lines.

Likewise, symplocamide A, isolated from Symploca

sp. showed potent cytotoxicity to lung cancer cells and

neuroblastoma cells (Linington et al. 2008).

Malyngamide 3 and cocosamides B are recently

isolated from L. majuscula and it showed weak

cytotoxicity against MCF7 breast cancer and HT-29

colon cancer cells (Gunasekera et al. 2011). Oftedal

et al. (2010) have screened several cyanobacteria for

the acute myeloid leukemia (AML), which is the

second most common form of leukemia. They have

found that the aqueous cyanobacterial extract is most

effective in causing apoptosis to AML cell lines. The

combination of a moderate concentration of the

anticancer drug daunorubicin with cyanobacterial

extract induced a synergistic apoptotic response in

AML cells. It can be concluded that these cyanobac-

terial apoptogens have the ability to greatly improve

the therapeutic index of daunorubicin (Oftedal et al.

2010). Also, there was no correlation between mouse

toxicity and induction of apoptosis neither in T cell

lymphoma nor in AML-cells (Oftedal et al. 2011).

Bisebromoamide, a new cell toxin that inhibits

cancer cell lines, was obtained from an Okinawan

948 Antonie van Leeuwenhoek (2013) 103:947961

123

Table 1 Bioactive molecules produced by various cyanobacteria

Bioactive molecules Cyanobacteria Bioactivity References

Symplocamide A Symploca sp. Anticancer Linington et al. (2008)

Symplostatin Symploca sp. Anticancer Luesch et al. (2002b)

Apratoxins Lyngbya majuscula, L. sordid,L. bouilloni

Anticancer Luesch et al. (2001a), Gutierrez

et al. (2008), Matthew et al. (2008),

Tidgewell et al. (2010)

Aplysiatoxin Lyngbya majuscula Anticancer Mynderse et al. (1977)

Lyngbyabellin B Lyngbya majuscula Anticancer Luesch et al. (2000)

Acutiphycin Oscillatoria acutissima Anticancer Barchi et al. (1984)

Dragonamide C, D Lyngbya polychroa Anticancer Gunasekera et al. (2008)

Cryptophycins Nostoc sp. Anticancer Moore et al. (1996)

Arenastatin A Dysidea arenaria Anticancer Moore et al. (1996)

Borophycin Nostoc linckia, N. spongiaeforme Anticancer Hemscheidt et al. (1994)

Homodolastatin 16 Lyngbya majuscula Anticancer Davies-Coleman et al. (2003)

Curacin A Lyngbya majuscula Anticancer Simmons et al. (2005)

Tjipanazoles Tolypothrix tjipanasensis Anticancer Bonjouklian et al. (1991)

Pitipeptolides A, B Lyngbya majuscula Anticancer Luesch et al. (2001c)

Aurilide Lyngbya majuscula Anticancer Han et al. (2006)

Carmabin A, B Lyngbya majuscula Anticancer McPhail et al. (2007)

Calothrixins A, B Calothrix sp. Anticancer Rickards et al. (1999)

Dolastatins Lyngbya sp., Symploca sp. Anticancer Fennell et al. (2003)

Biselyngbyaside Lyngbya sp. Anticancer Teruya et al. (2009b)

Ankaraholide A Geitlerinema sp. Anticancer Andrianasolo et al. (2005)

Malyngamide 3 Lyngbya majuscula Anticancer Gunasekera et al. (2011)

Cocosamides B Lyngbya majuscula Anticancer Gunasekera et al. (2011)

Bisebromoamide Lyngbya sp. Anticancer Teruya et al. (2009a)

Symplocamide A Symploca sp. Anticancer Linington et al. (2008)

Veraguamides Symploca cf. hydnoides Anticancer Salvador et al. (2011)

Largazole Symploca sp. Anticancer Taori et al. (2008)

C-phycocyanin Aphanizomenon flos-aquae Anticancer Tokuda et al. (1996)

Diacylglycerols Aphanizomenon flos-aquae Anticancer Tokuda et al. (1996)

Caylobolide Phormidium sp. Anticancer Salvador et al. (2010)

Coibamide Leptolyngbya sp. Anticancer Medina et al. (2008)

Hoiamide Association of Lyngbya majusculaand Phormidium gracile

Anticancer Choi et al. (2010)

Isomalyngamide Lyngbya majuscula Anticancer Chang et al. (2011)

Jamaicamides Lyngbya majuscula Anticancer Edwards et al. (2004)

Kalkitoxin Lyngbya majuscula Anticancer White et al. (2004)

Palauamide Lyngbya sp. Anticancer Zou et al. (2005)

Tasiamide Symploca sp. Anticancer Williams et al. (2003a)

Tasipeptins Symploca sp. Anticancer Williams et al. (2003b)

Wewakpeptins Lyngbya semiplena Anticancer Han et al. (2005)

Lagunamide Lyngbya majuscula Anticancer Tripathi et al. (2011)

Majusculamide Lyngbya majuscula Anticancer Pettit et al. (2008)

Malevamide Symploca hydnoides Anticancer Horgen et al. (2002)

Obyanamide Lyngbya confervoides Anticancer Williams et al. (2002)

Antonie van Leeuwenhoek (2013) 103:947961 949

123

collection of Lyngbya sp. (Teruya et al. 2009a). It

inhibits the phosphorylation of extracellular signal-

related protein kinase (ERK). Veraguamides from

cyanobacterium Symploca cf. hydnoides showed mod-

erate to weak cytotoxic activity against HT29 colo-

rectal adenocarcinoma and HeLa cervical carcinoma

cell lines (Salvador et al. 2011). Taori et al. (2008)

reported that largazole isolated from Symploca sp.

shows cytotoxicity against transformed mammary

epithelial cell lines (MDA-MB-231). The molecular

target for largazole is found to be histone deacetylases

(HDACs), and it is categorized as a class I HDAC

inhibitor.

Some workers have also reported the anticancerous

activity of photosynthetic pigment. Li et al. (2010)

reported anti-tumor activities of C-phycocyanin (C-

PC) mediated photodynamic therapy in MCF-7 breast

cell lines. Aphanizomenon flos-aquae extract contain-

ing a high concentration of phycocyanin inhibited the

in vitro growth of tumor cell lines, Phormidium tenui

contain several diacylglycerols that inhibit chemically

induced tumors in mice (Tokuda et al. 1996).

Recently, Gantar et al. (2012) reported that C-PC in

combination with lower dose (10 % of typical dose) of

anticancer drug topotecan can kill cancer cells at

higher rate than used alone at full dose.

Cyanobacteria cyclopeptides as a lead compound

for cancer treatment

Cyanobacterial cyclopeptides, microcystins (MCs)

and nodularins at high concentration are considered

to be toxic to humans (Carmichael et al. 2001; Funari

and Testai 2008). MCs are the most common cyano-

bacterial toxin prevalent in the water bodies. They can

be ingested through contaminated drinking water

(Oberholster et al. 2004), fish (Poste et al. 2011) or

sea foods (Mulvenna et al. 2012). From a pharmaco-

logical point of view, MCs are stable hydrophilic

Table 1 continued

Bioactive molecules Cyanobacteria Bioactivity References

Palmyramide Lyngbya majuscula Anticancer Taniguchi et al. (2010)

Ulongapeptin Lyngbya sp. Anticancer Williams et al. (2003c)

Grassypeptolide Lyngbya majuscula Anti-proliferative Kwan et al. (2008)

Nostoflan Nostoc flagelliforme Antiviral Kanekiyo et al. (2005)

Ichthyopeptins Microcystis ichthyoblabe Antiviral Zainuddin et al. (2007)

Bauerines AC Dichotrix baueriana Anti-HSV-2 Larsen et al. (1994)

Sulfolipids Lyngbya lagerhimii, Phormidium tenue Anti-HIV Gustafson et al. (1989)

Calcium spirulan Spirulina platensis Anti-HIV Hayashi et al. (1996)

Cyanovirin Nostoc ellipsosporum Anti-HIV Dey et al. (2000)

Scytovirin Scytonema varium Anti-HIV Xiong et al. (2006)

Sulfoglycolipid Scytonema sp. Anti-HIV Loya et al. (1998)

Ambiguines Fischerella sp. Antibacterial Raveh and Carmeli (2007)

Bastadin Anabaena basta Antibacterial Miao et al. (1990)

Bis-(v-butyrolactones) Anabena variabilis Antibacterial Ma and Led (2000)

Hapalindole Nostoc CCC537, Fischerella sp. Antibacterial Asthana et al. (2009)

Abietane diterpenes Microcoleous lacustris Antibacterial Gutierrez et al. (2008)

Nostocine A Nostoc spongiaeforme Antibacterial Hirata et al. (2003)

Noscomin Nostoc commune Antibacterial Jaki et al. (2000)

Didehydromirabazole Scytonema mirabile Antibacterial Stewart et al. (1988)

Tolyporphin Tolypothrix nodosa Antibacterial Prinsep et al. (1992)

Muscoride Nostoc muscorum Antibacterial Nagatsu et al. (1995)

Ambiguine Fischerella ambigua Antibacterial Raveh and Carmeli (2007)


123

cyclic heptapeptides causes cellular damage following

uptake via organic anion transporting polypeptides

(OATP) (Sainis et al. 2010; Fischera et al. 2005). Their

intracellular biological effects involve inhibition of

catalytic subunits of protein phosphatase 1 (PP1) and

PP2, glutathione depletion and generation of reactive

oxygen species (ROS) (Amado and Monserrat 2010).

However, there are certain OATPs which are prom-

inently expressed in cancer tissue as compared to

normal tissue, qualifying MCs as potential candidates

for cancer drug development (Sainis et al. 2010;

Monks et al. 2007).

In the era of targeted cancer therapy, cyanobacterial

toxins comprise a rich source of natural cytotoxic

compounds with a potential to target cancers express-

ing specific uptake transporters (Sainis et al. 2010).

Furthermore, a high proportion of these natural

products target eukaryotic cytoskeleton, such as

tubulin and actin microfilaments, making them an

attractive source of anticancer drugs (Tan 2010).

Antiviral compounds

The global spread of deadly viral diseases like HIV/

AIDS and avian influenza (H5N1 virus) etc. have

showed fatal consequences. Food and Drug Adminis-

tration (FDA) approved anti-HIV treatment highly

active antiretroviral therapy (HAART), is effective in

controlling the progression of HIV infections but

found to be toxic (Luescher-Mattli 2003). Thus, novel

drugs are now urgently required to combat deadliest

diseases. Antiviral compound isolated form cyano-

bacteria are usually found to show bioactivity by

blocking viral absorption or penetration and inhibiting

replication stages of progeny viruses after penetration

into cells. The protection of human lymphoblastoid T

cells from the cytopathic effect of HIV infection with

the extract of Lyngbya lagerheimeii and Phormidium

tenue has been reported by Gustafson et al. (1989). A

new class of HIV inhibitors called sulfonic acid,

containing glycolipid, was isolated from the extract of

cyanobacteria and the compounds were found to be

active against the HIV virus. Cyanovirin-N (CVN), a

peptide isolated from cyanobacteria, inactivates the

strains of HIV virus and inhibits cell to cell and virus to

cell fusion (Yang et al. 1997). In vitro and in vivo

antiviral tests suggested that the anti-HIV effect of

CVN is stronger than a well-known targeted (viral

entry) antibody (2G12) and another microbicide,

PRO2000 (Xiong et al. 2010).

Calcium spirulan (Ca-SP), a novel sulphated poly-

saccharide, is an antiviral agent. This compound

selectively inhibits the entry of enveloped virus

(Herpes simplex, humancytomegalo virus, measles

virus) into the cell (Hayashi et al. 1996). Rechter et al.

(2006) have analyzed polysaccharide fractions iso-

lated from Arthrospira platensis. These fractions

containing spirulan-like molecules showed a pro-

nounced antiviral activity against human cytomega-

lovirus, herpes simplex virus type 1.

Yakoot and Salem (2012) has conducted first

human trial to address the effect of Spirulina platensis

dried extract on virus load, liver function, health

related quality of life and sexual functions in patients

with chronic hepatitis C virus (HCV) infection. They

found the therapeutic potential of S. platensis in

chronic HCV patients, and in some cases (13 %) the

viral infection is complexly nullified. Mansour et al.

(2011) have found that the polysaccharides isolated

from Gloeocapsa turgidus and Synechococcus cedro-

rum showed higher antiviral activity against rabies

virus than that against herpes-1 virus. The exopoly-

saccharide from Aphanothece halophytica has an

antiviral activity against influenza virus A (H1N1),

which shows an 30 % inhibition of pneumonia in

infected mice (Zheng et al. 2006).

Antibacterial compounds

Noscomin from Nostoc commune exhibited antibac-

terial activity against Bacillus cereus, Staphylococcus

epidermidis, and Escherichia coli (Jaki et al. 2000).

Nostocarboline from Nostoc was found to inhibit the

growth of other cyanobacteria and green alga (Blom

et al. 2006). Hirata et al. (2003) found that nostocine A

isolated from Nostoc spongiaeforme exhibited growth

inhibitory stronger to green algae than to cyanobac-

teria. Asthana et al. (2009) have isolated hapalindole

(alkaloids) from Nostoc CCC537 and Fischerella sp.

and found antimicrobial activity against Mycobacte-

rium tuberculosis H37Rv, Staphylococcus aureus

ATCC25923, Salmonella typhi MTCC3216, Pseudo-

monas aeruginosa ATCC27853, E. coli ATCC25992

and Enterobacter aerogenes MTCC2822.

Ambiguine, a hapalindole-type alkaloid from Fi-

scherella ambigua shows antibacterial activity against


123

Cryptophycin 52 (C36H45ClN2O8 & 669.2 Da)

HN

O

O

H3C

O

ONHH3C CH3

O

O

O

H3 C

CH3Cl

OCH3

CH2S

CH3

N

H3CO

CH3

Curacin (C23H35NOS & 373.6 Da)

CH3

N

NH

N

N

CH3

CH3

H3C

O

H3C

H3C

O

H3C

CH3H3C

O

N

O

OO

H3C

H3C

N

O

O

OCH3

Dolastin 15 (C45H68N6O9 & 837 Da)

H3CN

HN

N

N

CH3

H3C CH3

OH3C CH3

O

CH3

CH3

CH 3

O

N

O

O

HN

H3C

CH 3

H3C

Tasidotin (C32H58N6O5 & 606.8 Da)

H3CN

HN

NN

CH3

H3C CH3

OH3C CH3

O

CH3

H3C CH3

OH O

NH

H3CO

CH3

O

Soblidotin (C36H61N5O6 & 660 Da)

H3C N

HN N

N

CH3

H3CCH3

OH3C

CH3

O

CH3

H3C

H3C

OCH3O NHH3CO

CH3

O

N

S

Dolastin 10 (C42H68N6O6S & 785 Da)

Cemadotin (C35H56N6O5 & 640.8 Da) H3C

N

NH

N

CH3

H3C

CH3 O

CH3

CH3

O

O

N

CH3

H3C

HN CH 3

O

O

N

Fig. 1 Structures of biomolecules which were in clinical trials


123

M. tuberculosis and against Bacillus anthracis (Raveh

and Carmeli 2007). Guo et al. (2009) have isolated

6-cyano-5-methoxy-12-methylindolo (2,3-a) carba-

zole from cyanobacteria and identified as a B. anthra-

cis inhibitor. The methanolic extract of S. platensis

showed broad spectrum antimicrobial activity and the

inhibition recorded was maximum for S. aureus

followed by E. coli, P. aeruginosa and S. typhi

(Kaushik and Chauhan 2008). Because of the growing

bacterial resistance against antibiotics and commercial

standard the search for new active substances with

antibacterial activity is urgently needed and cyano-

bacteria are the potential and promising candidates.

Cyanobacterial bioactive molecules under clinical

trials

The drug-development process normally proceeds

through various phases of clinical trials (phase 0 or

pre clinical, phase I, phase II and phase III). The FDA

must approve each phase before the study can

continue. Drugs are first tested in laboratory animal

(pre clinical phase) and in further phases healthy

human are tested. In phase I, II and III the number of

subjects range from 20 to 80, few dozen to 300 and

several hundred to 3,000 people, respectively. If the

drug successfully passes through all the phases it will

usually be approved by the National Regulatory

Authority (NRA) for use by the general population.

There are few prominent molecules from cyanobac-

teria such as dolastatins, cryptophycins, curacin and

their analogues which are in clinical trials as potential

anticancer drugs. The structures were drawn with the

help of ChemDraw (Fig. 1).

Dolastatins are the group of structurally unique

peptides which were first reported from marine animal

Dolabella auricularia (Pettit et al. 1987), using

microalgae as diet, but later on they were isolated

from the cyanobacteria Symploca (Luesch et al.

2001b). Dolastatins show anticancerous activity by

inhibiting microtubule assembly and many of its

analogues are in clinical trials. Till date there are

sixteen dolastatin forms isolated and are simply named

as dolastatin 1, 2, 3, and so on. Dolastatin 10 and 15 are

found to be showing promising results. National

Cancer Institute of US conducted the phase I clinical

trials of dolastatin 10 and progressed further to phase

II. Unfortunately, it was dropped from clinical trials,

due to some toxic effects (Simmons et al. 2005). This

finding results in the development of dolastatin 10

synthetic analogues like soblidotin, usually with

improved pharmacological and pharmacokinetic prop-

erties. Interestingly, its synthetic analogue soblidotin,

has cleared phase I and II successfully and now

undergoing phase III clinical trials under the supervi-

sion of Aska Pharmaceuticals, Tokyo, Japan (Bhatna-

gar and Kim 2010). The antitumor activity of

soblidotin, was found to be superior to existing

anticancer drugs, such as paclitaxel and vincristine

(Watanabe et al. 2006). The third generation dolastatin

15 analogues are cemadotin and tasidotin. Tasidotin is

antitumor agent and has cleared phase I trials (Mita

et al. 2006) and now undergoing phase II trials with

Genzyme Corporation, Cambridge, MA (Bhatnagar

and Kim 2010).

Cryptophycins are a group of cytotoxic depsipep-

tides, first isolated from Nostoc sp. as an antifungal

compound (Schwartz et al. 1990), later it was reported

to be effective against drug-resistant cancer cell lines

(Smith et al. 1994). Moore group (Chaganty et al.

2004; Golakoti et al. 1994, 1995) have isolated

twenty-six cryptophycins forms from Nostoc sp.

GSV 224. Of the various forms, cryptophycin 52

was found to be the most successful and evaluated in

phase II clinical trials for the treatment of platinum-

resistant ovarian cancer (DAgostino et al. 2006) and

advanced lung cancer (Edelman et al. 2003). Unfor-

tunately, the clinical trails were further discontinued

as it causes neuropathy and pain in the patients.

Magarvey et al. (2006) have analyzed cryptophycin

biosynthetic pathways that have opened more avenues

to create novel cryptophycin analogs (Sammet et al.

2010; Wei et al. 2012).

Another group, curacins are unique thiazoline-

containing lipopeptides that inhibits microtubule

assembly and it is a potent competitive inhibitor of

the binding of colchicine to tubulin (Blokhin et al.

1995). Gerwick group have isolated curacin A (Ger-

wick et al. 1994); curacin B and C (Yoo and Gerwick

1995); curacin D (Marquez et al. 1998) from Lyngbya

majuscule, which is prevalent in different water

bodies. Clinical development of curacin has been

hindered due to its low water solubility and thus it is

unable to produce activity during in vivo animal trails.

Hence, curacin was withdrawn from pre clinical

phase, but it served as a lead compound for the

development of synthetic analogs which are more


123

water soluble (Wipf et al. 2004). Isolation of all

of these compounds offer great opportunity and a

platform for the discovery of promising anticancer

agents.

Methods used for isolation and detection of novel

biomolecules

There are number of methods used for the extraction of

valuable metabolites. The extraction can either be

simultaneous (extra and intracellular metabolites) or

sequential (intercellular metabolites only). In both

types of extraction first quenching is done with liquid

nitrogen to freeze the metabolic activity. In simulta-

neous extraction, quenching is immediately followed

with extraction using a suitable solvent (butanol/

acetone/hexane/chloroform/methanol/aqueous metha-

nol/water/hexane/dichloromethane), while in sequen-

tial extraction biomass separation is done after

quenching and then various fractions are collected

by extracting with solvents of decreasing polarity

sequentially (water, aqueous methanol, methanol,

hexane) (Fig. 2). Conventional extraction methods

employed are solidliquid extraction (SLE) or liquid

liquid extraction (LLE). SLE is usually done by

soxhlet apparatus, it is the process of removing a

solutes from a solid (fixed phase) or matrix by using of

liquid solvent (mobile phase). Whereas in LLE, both

phase are in liquid phase and the separation of the

solute depend on the distribution coefficient of the

solute in mobile phase. Both SLE and LLE require

high volumes of solvents, long extraction times and

reproducibility of the results are low.

Recently low cost, chemical free green extraction

(GE) methods are used such as supercritical fluid

extraction (SFE), pressurized liquid extraction (PLE).

In SFE, the dissolved solutes are separated from the

raw material using certain gases (act as solvent) above

critical points. Carbon dioxide (CO2) gas is commonly

used solvent for the extraction of biomolecules. Some

of the researchers are also using ethanol as cosolvent

along with the CO2 gas. Mendiola et al. (2007) have

used SFECO2 (220 bar, 26.7 C) with 10 % ofethanol (cosolvent) for the extraction of antimicrobial

component from S. platensis. Onofrejova et al. (2010)

Simultaneous quenching (freeze metabolic activity) and extraction

with suitable solvent

Sequential sampling (Intracellular metabolites only)

Cyanobacterial Culture

Simultaneous sampling (Intracellular and extracellular

metabolites)

Quenching (to freeze metabolic activity)

Sequential extraction with various solvents

Biomass separation (by centrifugation and washing)

Fig. 2 Extraction procedure commonly followed for the isolation of novel biomolecules from cyanobacteria


123

have used combination of pressurized-liquid with

solid-phase extraction (PLESPE) for the isolation of

bioactive phenols freshwater algae.

There are number of analytical techniques available

for the detection and purification of biomolecules from

cyanobacterial extract (Fig. 3). Thin layer chromatog-

raphy (TLC) followed by spectrophotometric analysis

is the easiest technique used for primarily identifica-

tion and separation of bioactive molecules. Pelander

et al. (2000) have used high performance TLC plates

(HPTLC) for the separation of small cyanobacterial

peptides. However, TLC/HPTLC separation is non-

specific and less sensitive. Use of high performance

liquid chromatography (HPLC) for the identification

and quantification has increased greatly. Nowadays

more advance technique ultra performance liquid

chromatography (UPLC) is available, which can be

better option than HPLC. In order to get accurate

identification of bioactive product, liquid chromatog-

raphy is followed by mass spectrometry (LCMS)

(Harada et al. 2004). Different configurations of this

approach such as fast atom bombardment (FAB-LC

MS) (Kondo et al. 1995) and electrospray ionization

(ESI-LCMS) has been developed (Barco et al. 2002;

Spoof et al. 2003). Zhang et al. (2004) used LCMS

MS with electrospray ionization. Liquid chromatog-

raphy can also be coupled with quadruple time-of-

flight tandem mass spectrometry (LCQTof-MS) for

cyanobacterial cyclopeptides detection (Ferranti et al.

2009). Immunoassay techniques due to their high

sensitivity, specificity and operational simplicity are

widely used for the characterization of biomolecules.

Lindner et al. (2004) have developed the highly

sensitive enzyme-linked immunosorbent assays

(ELISA) for the detection and quantification of

cyanobacterial cyclopeptides.

Metabolites are also identified using matrix-

assisted laser desorption/ionization time-of-flight

mass spectrometry (MALDI-TOF MS). The technique

requires very small amount of sample without sepa-

ration or purification (Welker et al. 2002). Erhard et al.

(1997) used MALDI-TOF MS for identification of

secondary metabolites with intact cyanobacterial cells.

Resulting mass signals which are further characterized

by post source-decay fragmentation, and comparison

of observed fragment spectra with theoretical ones or

Cyanobacterial Extract

In Vivo assay

Chromatographic analysis

Mass spectral analysis Immunological assay

using ELISA

TLC/HPTLC

HPLC/UPLC-UV/PDA

Bioassay using bacterial/plant/animal system

In Vitro assay

GC-FID

MALDI-TOF MS

ESI-MS

DART-MS

DESI-MS

LC-MS

GC-MS

Fig. 3 Detection and purification methods for isolation of bioactive molecules from cyanbacterial extract


123

with those of pure reference compounds (Welker et al.

2004). In general, MALDI-TOF MS is used for the

identification of peptides; however this can also be

used for the identification of alkaloids (Araoz et al.

2008). In MALDI-TOF MS, the mass fragmentation

pattern of a particular analyte is distinctive, and may

vary according to the ionization mode used for mass

spectrometry and to the charge state of the molecule

(Antoine et al. 2006; Welker et al. 2006).

Desorption electrospray ionization mass spectrom-

etry (DESI-MS) is also an applied analytical technique

for chemical profiling, characterization and quantifi-

cation of low molecular-weight biomolecules

(Esquenazi et al. 2009). Another technique, direct

analysis in real time mass spectrometric (DART-MS)

technique is very much effective in chemical profiling

and fingerprinting of bioactive molecules without

prior sample preparation. Singh and Verma (2012)

have identified the Nostoc sp. on the basis of

characteristic chemical compounds (chemical finger-

printing) using DART-MS. All these techniques are

helpful for the identification and characterization of

bioactive molecules.

Conclusion

Cyanobacteria are the promising sources potentially

useful natural products. Microbial natural products

discovery opens a new era of research. Presently, the

isolation of number of natural products is increasing;

however, few compounds have reached the market.

Limited number of identified cyanobactererial bio-

molecules and analogues are in clinical trials and some

of them have passed different phases of clinical trials

to prove their candidature as potential drugs. In order

to exploit the new opportunities available, it will be

necessary to develop novel methodologies that allow

the isolation and culture of microorganisms, which

produce natural products unique to particular envi-

ronmental conditions. Thus, there is an urgent need for

extensive research in this new emerging field for drug

discovery.

Acknowledgments Authors are thankful to Director, CSIR-NBRI for all the facilities and constant encouragement. Rakhi

Bajpai Dixit is grateful to Department of Science and Technology

(DST, New Delhi), for providing financial assistance in the form

of a project (Ref. No. SR/FT/LS-111/2010).

References

Amado LL, Monserrat JM (2010) Oxidative stress generation by

microcystins in aquatic animals: why and how. Environ Int

36:226235

Andrianasolo EH, Gross H, Goeger D, Musafija-Girt M, McP-

hail K, Leal RM, Mooberry SL, Gerwick WH (2005) Iso-

lation of swinholide A and related glycosylated derivatives

from two field collections of marine cyanobacteria. Org

Lett 7:13751378

Antoine R, Broyer M, Chamot-Rooke J, Dedonder C, Desfr-

ancois C, Dugourd P, Gregoire G, Jouvet C, Onidas D,

Poulain P, Tabarin T, van der Rest G (2006) Comparison of

the fragmentation pattern induced by collisions, laser

excitation and electron capture. Influence of the initial

excitation. Rapid Commun Mass Spectrom 20:16481652

Araoz R, Guerineau V, Rippka R, Palibroda N, Herdman M,

Laprevote O, von Dohren H, Tandeau de Marsac N, Erhard

M (2008) MALDI-TOF-MS detection of the low molecular

weight neurotoxins anatoxin-a and homoanatoxin-a on

lyophilized and fresh filaments of axenic Oscillatoriastrains. Toxicon 51:13081315

Asthana RK, Tripathi MK, Deepali A, Srivastava A, Singh AP,

Singh SP, Nath G, Srivastava R, Srivastava BS (2009)

Isolation and identification of a new antibacterial entity

from the Antarctic cyanobacterium Nostoc CCC 537.J Appl Phycol 21:8188

Bajpai R, Sharma NK, Rai AK (2010) Anticancerous/antiviral

compounds from cyanobacteria. Algas 43:46

Barchi JJ, Moore RE, Patterson GML (1984) Acutiphycin and

20, 21-didehydroacutiphycin, new antineoplastic agents

from the cyanophyte Oscillatoria acutissima. J Am Chem

Soc 106:81938197

Barco M, Rivera J, Caixach J (2002) Analysis of cyanobacterial

hepatotoxins in water samples by microbore reversed-

phase liquid chromatographyelectrospray ionisation mass

spectrometry. J Chromatogr A 959:103111

Blokhin AV, Yoo H-D, Geralds RS, Nagle DG, Gerwick WH,

Hamel E (1995) Characterization of the interaction of the

marine cyanobacterial natural product curacin A with the

colchicine site of tubulin and initial structure-activity

studies with analogs. Mol Pharmacol 48:523531

Blom JF, Brutsch T, Barbaras D, Bethuel Y, Locher HH, Hub-

schwerlen C, Gademann K (2006) Potent algicides based

on the cyanobacterial alkaloid nostocarboline. Org Lett

8:737740

Bonjouklian R, Smitka TA, Doolin LE, Molloy RM, Debono M,

Shaffer SA, Moore RE, Stewart JB, Patterson GML (1991)

Tjipanazoles, new antifungal agents from the blue-green

alga Tolypothrix tjipanasensis. Tetrahedron 47:77397750Burja AM, Banaigs B, Abou-Mansour E, Burgess G, Wright PC

(2001) Marine cyanobacteria: a prolific source of natural

products. Tetrahedron 57:93479377

Cardellina JH, Moore BS (2010) Editorial: Richard E Moore

(19332007). J Nat Prod 73:301302

Carmichael WW, Azevedo SMFO, Ji SA, Molica RJR, Jo-

chimsen EM, Lau S, Rinehart KL, Shaw GR, Eaglesham

GK (2001) Human fatalities from cyanobacteria: chemical

and biological evidence for cyanotoxins. Environ Health

Perspect 109:663668


123

Catassi A, Cesario A, Arzani D, Menichini P, Alama A, Bruzzo

C, Imperatori A, Rotolo N, Granone P, Russo P (2006)

Characterization of apoptosis induced by marine natural

products in non small cell lung cancer A549 cells. Cell Mol

Life Sci 63:23772386

Chaganty S, Golakoti T, Heltzel C, Moore RE, Yoshida WY

(2004) Isolation and structure determination of crypto-

phycins 38, 326, and 327 from the terrestrial cyanobacte-

rium Nostoc sp. GSV 224. J Nat Prod 67:14031406Chang TT, More SV, Lu IH, Hsu JC, Chen TJ, Jen YC, Lu CK,

Li WS (2011) Isomalyngamide A, A-1 and their analogs

suppress cancer cell migration in vitro. Eur J Med Chem

46:38103819

Choi H, Pereira AR, Cao Z, Shuman CF, Engene N, Byrum T,

Matainaho T, Murray TF, Mangoni A, Gerwick WH (2010)

The hoiamides, structurally intriguing neurotoxic lipo-

peptides from Papua New Guinea marine cyanobacteria.

J Nat Prod 73:14111421

Chorus I, Bartram J (eds) (1999) Toxic cyanobacteria in water, a

guide of their public health consequences, monitoring, and

management. WHO E and FN Spoon London

DAgostino G, Del Campo J, Mellado B, Izquierdo MA, Mi-

narik T, Cirri L, Marini L, Perez- Gracia JL, Scambia G

(2006) A Multicenter phase ii study of the cryptophycin

analog LY355703 in patients with platinum-resistant

ovarian cancer. Int J Gynecol Cancer 16:7176

Davies-Coleman M, Dzeha TM, Gray CA, Hess S, Pannell LK,

Hendricks DT, Arendse CE (2003) Isolation of homodo-

lastatin 16, a new cyclic depsipeptide from a Kenyan col-

lection of Lyngbya majuscula. J Nat Prod 66:712715Dey B, Lerner DL, Lusso P, Boyd MR, Elder JH, Berger EA

(2000) Multiple antiviral activities of cyanovirin-N:

blocking of human immunodeficiency virus type 1 gp120

interaction with CD4 and coreceptor and inhibition of

diverse enveloped viruses. J Virol 74:45624569

Edelman MJ, Gandara DR, Hausner P, Israel V, Thornton D,

DeSanto J, Doyle LA (2003) Phase 2 study of cryptophycin

52 (LY355703) in patients previously treated with plati-

num based chemotherapy for advanced non-small cell lung

cancer. Lung Cancer 39:197199

Edwards DJ, Marquez BL, Nogle LM, McPhail K, Goeger DE,

Roberts MA, Gerwick WH (2004) Structure and biosyn-

thesis of the jamaicamides, new mixed polyketide-peptide

neurotoxins from the marine cyanobacterium Lyngbyamajuscula. Chem Biol 11:817833

Erhard M, von Dohren H, Jungblut P (1997) Rapid typing and

elucidation of new secondary metabolites of intact cya-

nobacteria using MALDI-TOF mass spectrometry. Nat

Biotechnol 15:906909

Esquenazi Eduardo, Dorrestein Pieter C, Gerwick William H

(2009) Probing marine natural product defenses with

DESI-imaging mass spectrometry. Proc Natl Acad Sci

USA 106:72697270

Fennell BJ, Carolan S, Pettit GR, Bell A (2003) Effects of the

antimitotic natural product dolastatin 10, and related pep-

tides, on the human malarial parasite Plasmodium falci-parum. J Antimicrob Chemother 51:833841

Ferranti P, Fabbrocino S, Nasi A, Caira S, Bruno M, Serpe L,

Gallo P (2009) Liquid chromatography coupled to qua-

druple time-of-flight tandem mass spectrometry for micr-

ocystin analysis in freshwaters: method performances and

characterisation of a novel variant of microcystin-RR.

Rapid Commun Mass Spectrom 23:13281336

Fischera WJ, Altheimera S, Cattorib V, Meierb PJ, Dietricha

DR, Hagenbuchb B (2005) Organic anion transporting

polypeptides expressed in liver and brain mediate uptake of

microcystin. Toxicol Appl Pharmacol 203:257263

Funari E, Testai E (2008) Human health risk assessment related

to cyanotoxins exposure. Crit Rev Toxicol 38:97125

Gademann K, Portmann C (2008) Secondary metabolites from

cyanobacteria: complex structures and powerful bioactiv-

ities. Curr Org Chem 12:326341

Gantar M, Dhandayuthapani S, Rathinavelu A (2012) Phycocy-

anin induces apoptosis and enhances the effect of topotecan

on prostate cell line LNCaP. J Med Food 15:10911095

Gerwick WH, Proteau PJ, Nagle DG, Hamel E, Blokhin A, Slate

DL (1994) Structure of curacin A, a novel antimitotic,

antiproliferative, and brine shrimp toxic natural product

from the marine cyanobacterium Lyngbya majuscula. J OrgChem 59:12431245

Gerwick WH, Coates RC, Engene N, Gerwick L, Grindberg RV,

Jones AC, Sorrels CM (2008) Giant marine cyanobacteria

produce exciting potential pharmaceuticals. Microbe

3:277284

Golakoti T, Ohtani I, Patterson GML, Moore RE, Corbett TH,

Valeriote FA, Demchik L (1994) Total structures of

cryptophycins, potent antitumor depsipeptides from the

blue-green-alga Nostoc sp. Strain GSV-224. J Am ChemSoc 116:47294737

Golakoti T, Ogino J, Heltzel CE, Lehusebo T, Jensen CM,

Larsen LK, Patterson GML, Moore RE, Mooberry SL,

Corbett TH, Valeriote FA (1995) Structure determination,

conformational-analysis, chemical-stability studies, and

antitumor evaluation of the cryptophycins: isolation of 18

new analogs from Nostoc sp. Strain GSV-224. J Am ChemSoc 117:1203012049

Gunasekera SP, Ross C, Paul VJ, Matthew S, Luesch H (2008)

Dragonamides C and D, linear lipopeptides from the

marine cyanobacterium brown Lyngbya polychroa. J NatProd 71:887890

Gunasekera SP, Owle CS, Montaser R, Luesch H, Paul VJ

(2011) Malyngamide 3 and cocosamides A and B from the

marine cyanobacterium Lyngbya majuscula from CocosLagoon, Guam. J Nat Prod 74:871876

Guo S, Tipparaju SK, Pegan SD, Wan B, Mo S, Orjala J,

Mesecar AD, Franzblau SG, Kozikowski AP (2009) Nat-

ural product leads for drug discovery: isolation, synthesis

and biological evaluation of 6-cyano-5-methoxyindo-

lo[2,3-a]carbazole based ligands as antibacterial agents.

Bioorg Med Chem 17:71267130

Gustafson KR, Cardellina JH, Fuller RW, Wieslow OS, Kiser

RF, Sander KM, Patterson GM, Boyd MR (1989) AIDS

antiviral sulfolipids from cyanobacteria (blue-green algae).

J Natl Cancer Inst 81:12541258

Gutierrez M, Suyama TL, Engene N, Wingerd JS, Matainaho T,

Gerwick WH (2008) Apratoxin D, a potent cytotoxic cy-

clodepsipeptide from Papua New Guinea collections of the

marine cyanobacteria Lyngbya majuscula and Lyngbyasordida. J Nat Prod 71:10991103

Gutierrez RMP, Flores AM, Solis RV, Jimenez JC (2008) Two

new antibacterial norbietane diterpenoids from cyanobac-

terium Micrococcus lacustris. J Nat Med 62:328331


123

Han B, Goeger D, Maier CS, Gerwick WH (2005) The we-

wakpeptins, cyclic depsipeptides from a Papua New Gui-

nea collection of the marine cyanobacterium Lyngbyasemiplena. J Org Chem 70:31333139

Han B, Gross H, Goeger DE, Mooberry SL, Gerwick WH (2006)

Aurilides B and C, cancer cell toxins from a Papua New

Guinea collection of the marine cyanobacterium Lyngbyamajuscula. J Nat Prod 69:572575

Harada K-I, Nakano T, Fujii K, Shirai M (2004) Comprehensive

analysis system using liquid chromatographymass spec-

trometry for the biosynthetic study of peptides produced by

the cyanobacteria. J Chromatogr A 1033:107113

Hayashi K, Hayashi T, Kojima IA (1996) Natural sulfated

polysaccharide, calcium spirulan, isolated from Spirulinaplatensis: in vitro and ex vivo evaluation of anti-herpessimplex virus and anti-human immunodeficiency virus

activities. AIDS Res Hum Retroviruses 12:14631471

Hemscheidt T, Puglisi MP, Larsen LK, Patterson GML, Moore

RE, Rios JL, Clardy J (1994) Structure and biosynthesis of

borophycin, a new boeseken complex of boric acid from a

marine strain of the blue-green alga Nostoc linckia. J OrgChem 59:34673471

Hirata K, Yoshitomi S, Dwi S, Iwabe O, Mahakhant A, Polchai

J, Miyamoto K (2003) Bioactivities of nostocine A pro-

duced by a freshwater cyanobacterium Nostoc spongiae-forme TISTR 8169. J Biosci Bioeng 95:512517

Horgen FD, Kazmierski EB, Westenburg HE, Yoshida WY,

Scheuer PJ (2002) Malevamide D: isolation and structure

determination of an isodolastatin H analogue from the

marine cyanobacterium Symploca hydnoides. J Nat Prod65:487491

Ira Bhatnagar, Kim S-K (2010) Immense essence of excellence:

marine microbial bioactive compounds. Mar Drugs

8:26732701

Jaki B, Orjala J, Heilmann J, Linden A, Vogler B, Sticher O

(2000) Novel extracellular diterpenoids with biological

activity from the cyanobacterium Nostoc commune. J NatProd 63:339343

Jordan MA, Wilson L (1998) Microtubules and actin filaments:

dynamic targets for cancer chemotherapy. Curr Opin Cell

Biol 10:123130

Kalemkerian GP, Ou XL, Adil MR, Rosati R, Khoulani MM,

Madan SK, Pettit GR (1999) Activity of dolastatin 10

against small-cell lung cancer in vitro and in vivo: induc-

tion of apoptosis and bcl-2 modification. Cancer Chemo-

ther Pharm 43:507515

Kanekiyo K, Lee JB, Hayashi K, Takenaka H, Hayakawa Y,

Endo S, Hayashi T (2005) Isolation of an antiviral poly-

saccharide, nostoflan, from a terrestrial cyanobacteium,

Nostoc flagilliforme. J Nat Prod 68:10371041Kaushik P, Chauhan A (2008) In vitro antibacterial activity of

laboratory grown culture of Spirulina platensis. Indian JMicrobiol 48:348352

Kondo F, Ikai Y, Oka H, Matsumoto H, Yamada S, Ishikawa N,

Tsuji K, Harada K-I, Shimada T, Oshikata M, Suzuki M

(1995) Reliable and sensitive method for determination of

microcystins in complicated matrices by frit-fast atom

bombardment liquid chromatography/mass spectrometry.

Nat Toxins 3:4149

Kwan JC, Rocca JR, Abboud KA, Paul VJ, Luesch H (2008)

Total structure determination of grassypeptolide, a new

marine cyanobacterial cytotoxin. Org Lett 10:789792

Larsen LK, Moore RE, Patterson GML (1994) Beta-carbolines

from the blue-green alga Dichothrix baueriana. J Nat Prod57:419421

Li B, Chu X, Gao M, Li W (2010) Apoptotic mechanism of

MCF-7 breast cells in vivo and in vitro induced by pho-

todynamic therapy with C-phycocyanin. Acta Biochim

Biophys Sin 42:8089

Lindner P, Molz R, Yacoub-George E, Durkop A, Wolf H

(2004) Development of a highly sensitive inhibition

immunoassay for microcystin-LR. Anal Chim Acta

521:3744

Linington RG, Edwards DJ, Shuman CF, McPhail KL, Matai-

naho T, Gerwick WH (2008) Symplocamide A, a potent

cytotoxin and chymotrypsin inhibitor from the marine

cyanobacterium Symploca sp. J Nat Prod 71:2227Loya S, Reshef V, Mizrachi E, Silberstein C, Rachamim Y,

Carmeli S, Hizi A (1998) The inhibition of the reverse

transcriptase of HIV-1 by the natural sulfoglycolipids from

cyanobacteria: contribution of different moieties to their

high potency. J Nat Prod 61:891895

Luesch H, Yoshida WY, Moore RE, Paul VJ, Mooberry SL

(2000) Isolation, structure determination, and biological

activity of Lyngbyabellin A from the marine cyanobacte-

rium Lyngbya majuscula. J Nat Prod 63:611615Luesch H, Moore RE, Paul VJ, Mooberry SL, Corbett TH

(2001a) Isolation of dolastatin 10 from the marine cyano-

bacterium Symploca species VP642 and total stereochem-istry and biological evaluation of its analogue symplostatin

1. J Nat Prod 64:907910

Luesch H, Pangilinan R, Yoshida WY, Moore RE, Paul VJ

(2001b) Pitipeptolides A and B, new cyclodepsipeptides

from the marine cyanobacterium Lyngbya majuscula. J NatProd 64:304307

Luesch H, Yoshida WY, Moore RE, Paul VJ, Corbett TH

(2001c) Total structure determination of apratoxin A, a

potent novel cytotoxin from the marine cyanobacterium

Lyngbya majuscula. J Am Chem Soc 123:54185423Luesch H, Yoshida WY, Moore RE, Paul VJ (2002a) New

apratoxins of marine cyanobacterial origin from Guam and

Palau. Bioorg Med Chem 10:19731978

Luesch H, Yoshida WY, Moore RE, Paul VJ, Mooberry SL,

Corbett TH (2002b) Symplostatin 3, a new dolastatin 10

analogue from the marine cyanobacterium Symploca sp.VP452. J Nat Prod 65:1620

Luescher-Mattli M (2003) Algae as a possible source of new

antiviral agents. Curr Med Chem Anti-infect Agents

2:219225

Ma LX, Led JJ (2000) Determination by high field NMR

spectroscopy of the longitudinal electron relaxation rate in

Cu (II) plastocyanin form Anabaena variabilis. Am ChemSoc 122:78237824

Magarvey NA, Beck ZQ, Golakoti T, Ding Y, Huber U,

Hemscheidt TK, Abelson D, Moore RE, Sherman DH

(2006) Biosynthetic characterization and chemoenzymatic

assembly of the cryptophycins. Potent anticancer agents

from cyanobionts. ACS Chem Biol 1:766779


123

Mansour HA, Shoman SA, Kdodier MH (2011) Antiviral effect

of edaphic cyanophytes on rabies and herpes-1 viruses.

Acta Biol Hung 62:194203

Marquez B, Verdier-Pinard P, Hamel E, Gerwick WH (1998)

Curacin D, an antimitotic agent from the marine cyanobac-

terium Lyngbya majuscula. Phytochemistry 49:23872389Matthew S, Schupp PJ, Luesch H (2008) Apratoxin E, a cytotoxic

peptolide from a Guamanian collection of the marine cya-

nobacterium Lyngbya bouillonii. J Nat Prod 71:11131116McPhail KL, Correa J, Linington RG, Gonzalez J, Ortega-Barra

E, Capson TL, Gerwick WH (2007) Antimalarial linear

lipopeptides from a Panamanian strain of the marine cya-

nobacterium Lyngbya majuscule. J Nat Prod 70:984988Medina RA, Goeger DE, Hills P, Mooberry SL, Huang N, Ro-

mero LI, Ortega-Barria E, Gerwick WH, McPhail KL

(2008) Coibamide A, a potent antiproliferative cyclic

depsipeptide from the Panamanian marine cyanobacterium

Leptolyngbya sp. J Am Chem Soc 130:63246325Mendiola JA, Jaime L, Santoyo S, Reglero G, Cifuentes A,

Ibanez E, Senorans FJ (2007) Screening of functional

compounds in supercritical fluid extracts from Spirulinaplatensis. Food Chem 102:13571367

Miao S, Anderson RJ, Allen TM (1990) Cytotoxic metabolites

from the sponge Ianthella basta collected in Papua NewGuinea. J Nat Prod 53:14411446

Mita AC, Hammond LA, Bonate PL, Weiss G, McCreery H,

Syed S, Garrison M, Chu QS, DeBono JS, Jones CB,

Weitman S, Rowinsky EK (2006) Phase I and pharmaco-

kinetic study of tasidotin hydrochloride (ILX651), a third-

generation dolastatin-15 analogues, administered weekly

for 3 weeks every 28 days in patients with advanced solid

tumors. Clin Cancer Res 12:52075215

Monks NR, Liu S, Xu Y, Yu H, Bendelow AS, Moscow JA

(2007) Potent toxicity of the phosphatase inhibitor micro-

cystin LR and microcystin analogues in OATPB1- and

OATP1B3-expressing HeLa cells. Mol Cancer Ther

6:587598

Moore RE, Corbett TH, Patterson GML, Valeriote FA (1996)

The search for new antitumor drugs from blue green algae.

Current Pharm Design 2:317330

Muir JC, Pattenden G, Ye T (2002) Total synthesis of (?)-

curacin A, a novel antimitotic metabolite from a cyano-

bacterium. J Chem Soc Perkin Trans 1:22432250

Mulvenna V, Dale K, Priestly B, Mueller U, Humpage A, Shaw

G, Allinson G, Ian Falconer (2012) Health risk assessment

for cyanobacterial toxins in seafood. Int J Environ Res

Public Health 9:807820

Mynderse JS, Moore RE, Kashiwagi M, Norton TR (1977)

Antileukemia activity in the Oscillatoriaceae: isolation of

debromoaplysiatoxin from Lyngbya. Science 196:538540Nagatsu A, Kajitani H, Sakakibara J (1995) Muscoride A: a new

oxazole peptide alkaloid from freshwater cyanobacterium

Nostoc muscorum. Tetrahedron Lett 36:40974100Nagle DG, Geralds RS, Yoo H, Gerwick WH (1995) Absolute

configuration of curacin A, a novel antimitotic agent from

the tropical marine cyanobacterium Lyngbya majuscula.Tetrahedron Lett 36:11891192

Nunnery JK, Mevers E, Gerwick WH (2010) Biologically active

secondary metabolites from marine cyanobacteria. Curr

Opin Biotechnol 21:787793

Oberholster PJ, Botha A-M, Grobbelaar JU (2004) Microcystisaeruginosa: source of toxic microcystins in drinking water.Afr J Biotechnol 3:159168

Oftedal L, Selheim F, Wahlsten M, Sivonen K, Dskeland SO,

Herfindal L (2010) Marine benthic cyanobacteria contain

apoptosis-inducing activity synergizing with daunorubicin

to kill leukemia cells, but not cardiomyocytes. Mar Drugs

8:26592672

Oftedal L, Skjrven KH, Coyne RT, Edvardsen B, Rohrlack T,

Skulberg OM, Dskeland SO, Herfindal L (2011) The

apoptosis-inducing activity towards leukemia and lym-

phoma cells in a cyanobacterial culture collection is not

associated with mouse bioassay toxicity. J Ind Microbiol

Biotechnol 38:489501

Onofrejova L, Vasckova J, Klejdus B, Stratil P, Misurcova L,

Kracmar S, Kopecky J, Vacek J (2010) Bioactive phenols

in algae: the application of pressurized-liquid and solid-

phase extraction techniques. J Pharm Biomed Anal

51:464470

Pelander A, Ojanperae` I, Lahti K, Niinivaara K, Vuori E (2000)

Visual detection of cyanobacterial hepatotoxins by thin-

layer chromatography and application to water analysis.

Wat Res 34:26432652

Pettit GR, Kamano Y, Herald CL, Tuinman AA, Boettner FE,

Kizu H, Schmidt JM, Baczynskyj L, Tomer KB, Bontems

RJ (1987) The isolation and structure of a remarkable

marine animal antineoplastic constituent: dolastatin 10.

J Am Chem Soc 109:68836885

Pettit GR, Hogan F, Xu JP, Tan R, Nogawa T, Cichacz Z, Pettit

RK, Du J, Ye QH, Cragg GM, Herald CL, Hoard MS,

Goswami A, Searcy J, Tackett L, Doubek DL, Williams L,

Hooper JN, Schmidt JM, Chapuis JC, Tackett DN, Craci-

unescu F (2008) Antineoplastic agents. 536. New sources

of naturally occurring cancer cell growth inhibitors from

marine organisms, terrestrial plants, and microorganisms

(1a,). J Nat Prod 71:438444

Poste AE, Hecky RE, Guildford SJ (2011) Evaluating micro-

cystin exposure risk through fish consumption. Environ Sci

Technol 45:58065811

Prinsep MR, Caplan FR, Moore RE, Patterson GML, Smith CD

(1992) Tolyphorin, a novel multidrug resistance reversing

agent from the blue green algae Tolypothrix nodosa. J AmChem Soc 114:385387

Raveh A, Carmeli S (2007) Antimicrobial ambiguines from the

cyanobacterium Fischerella sp collected in Israel. J NatProd 70:196201

Rechter S, Konig T, Auerochs S, Thulke S, Walter H, Dornen-

burg H, Walter C, Marschall M (2006) Antiviral activity of

Arthrospira-derived spirulan-like substances. AntiviralRes 72:197206

Rickards RW, Rothschild JM, Willis AC, de Chazal NM, Kirk J,

Kirk K, Saliba KJ, Smith GD (1999) Calothrixins A and B,

novel pentacyclic metabolites from Calothrix cyanobac-teria with potent activity against malaria parasites and

human cancer cells. Tetrahedron 55:1351313520

Sainis I, Fokas D, Vareli K, Tzakos AG, Kounnis V, Briasoulis

E (2010) Cyanobacterial cyclopeptides as lead compounds

to novel targeted cancer drugs. Mar Drugs 8:629657

Salvador LA, Paul VJ, Luesch H (2010) Caylobolide B, a

macrolactone from symplostatin 1-producing marine


123

cyanobacteria Phormidium spp. from Florida. J Nat Prod73:16061609

Salvador LA, Biggs JS, Paul VJ, Luesch H (2011) Veragua-

mides AG, cyclic hexa depsipeptides from a dolastatin

16-producing cyanobacterium Symploca cf hydnoides

from Guam. J Nat Prod 74:917927

Sammet B, Bogner T, Nahrwold M, Weiss C, Sewald N (2010)

Approaches for the synthesis of functionalized crypto-

phycins. J Org Chem 75:69536960

Schwartz RE, Hirsch CF, Sesin DF, Flor JE, Chartrain M,

Fromtling RE, Harris GH, Salvatore MJ, Liesch JM, Yudin

K (1990) Pharmaceuticals from cultured algae. J Ind

Microbiol 5:113123

Schwarzer D, Finking R, Marahiel MA (2003) Nonribosomal

peptides: from genes to products. Nat Prod Rep 20:

275287

Simmons TL, Andrianasolo E, McPhail K, Flatt P, Gerwick WH

(2005) Marine natural products as anticancer drugs. Mol

Cancer Ther 4:333342

Singh S, Verma SK (2012) Application of direct analysis in real

time mass spectrometry (DART-MS) for identification of

an epiphytic cyanobacterium, Nostoc sp. Anal Lett 45:25622568

Singh S, Kate BN, Banerjee UC (2005) Bioactive compounds

from cyanobacteria and microalgae: an overview. Crit Rev

Biotechnol 25:7395

Singh RK, Tiwari SP, Rai AK, Mohapatra TM (2011) Cyano-

bacteria: an emerging source for drug discovery. J Antibiot

64:401412

Smith CD, Zhang X, Mooberry SL, Patterson GML, Moore RE

(1994) Cryptophycin: a new antimicrotubule agent active

against drug-resistant cells. Cancer Res 54:37793784

Spoof L, Vesterkvist P, Lindholm T, Meriluoto J (2003)

Screening for cyanobacterial hepatotoxins, microcystins

and nodularin in environmental water samples by reversed-

phase liquid chromatographyelectrospray ionisation mass

spectrometry. J Chromatogr A 1020:105119

Stewart JB, Bomemann V, Chen JL, Moore RE, Caplan FR,

Karuso H, Larsen LK, Patterson GM (1988) Cytotoxic,

fungicidal nucleosides from blue-green algae belonging to

the Scytonemataceae. J Antibiot 41:10481056

Tan LT (2007) Bioactive natural products from marine

cyanobacteria for drug discovery. Phytochemistry 68:

954979

Tan LK (2010) Filamentous tropical marine cyanobacteria: a

rich source of natural products for anticancer drug dis-

covery. J Appl Phycol 22:659676

Taniguchi M, Nunnery JK, Engene N, Esquenazi E, Byrum T,

Dorrestein PC, Gerwick WH (2010) Palmyramide A, a

cyclic depsipeptide from a Palmyra Atoll collection of the

marine cyanobacterium Lyngbya majuscula. J Nat Prod73:393398

Taori K, Paul VJ, Luesch H (2008) Structure and activity of

largazole, a potent antiproliferative agent from the Flo-

ridian marine cyanobacterium Symploca sp. J Am ChemSoc 130:18061807

Teruya T, Sasaki H, Fukazawa H, Suenaga K (2009a) Bise-

bromoamide, a potent cytotoxic peptide from the marine

cyanobacterium Lyngbya sp.: isolation, stereostructure,and biological activity. Org Lett 11:50625065

Teruya T, Sasaki H, Kitamura K, Nakayama T, Suenaga K

(2009b) Biselyngbyaside, a macrolide glycoside from the

marine cyanobacterium Lyngbya sp. Org Lett 11:24212424Tidgewell K, Engene N, Byrum T, Media J, Doi T, Valeriote

FA, Gerwick WH (2010) Evolved diversification of a

modular natural product pathway: apratoxins F and G, two

cytotoxic cyclic depsipeptides from a Palmyra collection of

Lyngbya bouillonii. Chem Bio Chem 11:14581466Tokuda H, Nishino H, Shirahashi H, Murakami N, Nagatsu A,

Sakakibara J (1996) Inhibition of 12-O-tetradecanoyl-phorbol-13-acetate promoted mouse skin papilloma by

digalactosyl diacylglycerols from the fresh water cyano-

bacterium Phormidium tenue. Cancer Lett 104:9195Tripathi A, Puddick J, Prinsep MR, Rottmann M, Chan KP,

Chen DY, Tan LT (2011) Lagunamide C, a cytotoxic cy-

clodepsipeptide from the marine cyanobacterium Lyngbyamajuscula. Phytochemistry 72:23692375

Watanabe J, Minami M, Kobayashi M (2006) Antitumor

activity of TZT-1027 (soblidotin). Anticancer Res

26:19731981

Wei C, Bogner T, Sammet B, Sewald N (2012) Total synthesis

and biological evaluation of fluorinated cryptophycins.

Beilstein J Org Chem 8:20602066

Welker M, Fastner J, Erhard M, von Dohren H (2002) Appli-

cation of MALDI-TOF MS in cyanotoxin research. Envi-

ron Toxicol 17:367374

Welker M, Brunke M, Preussel K, Lippert I, Dohren H (2004)

Diversity and distribution of Microcystis (Cyanobacteria)oligopeptide chemotypes from natural communities stud-

ied by single-colony mass spectrometry. Microbiology

150:17851796

Welker M, Marsalek B, Sejnohova L, von Dohren H (2006)

Detection and identification of oligopeptides in Microcys-tis (cyanobacteria) colonies: toward an understanding ofmetabolic diversity. Peptides 27:20902103

White JD, Xu Q, Lee CS, Valeriote FA (2004) Total synthesis

and biological evaluation of (?)-kalkitoxin, a cytotoxic

metabolite of the cyanobacterium Lyngbya majuscula. OrgBiomol Chem 2:20922102

Williams PG, Yoshida WY, Moore RE, Paul VJ (2002) Isolation

and structure determination of obyanamide, a novel cyto-

toxic cyclic depsipeptide from the marine cyanobacterium

Lyngbya confervoides. J Nat Prod 65:2931Williams PG, Yoshida WY, Moore RE, Paul VJ (2003a) The

isolation and structure elucidation of tasiamide B, a

4-amino-3-hydroxy-5-phenylpentanoic acid containing

peptide from the marine cyanobacterium Symploca sp.J Nat Prod 66:10061009

Williams PG, Yoshida WY, Moore RE, Paul VJ (2003b) Tasi-

peptins A and B: new cytotoxic depsipeptides from the

marine cyanobacterium Symploca sp. J Nat Prod 66:620624Williams PG, Yoshida WY, Quon MK, Moore RE, Paul VJ

(2003c) Ulongapeptin, a cytotoxic cyclic depsipeptide

from a Palauan marine cyanobacterium Lyngbya sp. J NatProd 66:651654

Wipf P, Reeves JT, Day BW (2004) Chemistry and biology of

curacin A. Curr Pharm Des 10:14171437

Xiong C, OKeefe BR, Byrd RA, McMohan JB (2006) Potent

anti-HIV activity of scytovirin domain 1 peptide. Peptides

27:16681675


123

Xiong S, Fan J, Kitazato K (2010) The antiviral protein cya-

novirin-N: the current state of its production and applica-

tions. Appl Microbiol Biotechnol 86:805812

Yakoot M, Salem A (2012) Spirulina platensis versus silymarinin the treatment of chronic hepatitis C virus infection. A

pilot randomized, comparative clinical trial. BMC Gas-

troenterol 12:32

Yang H, Lee E, Kim H (1997) Spirulina platensis inhibitsanaphylactic reaction. Life Sci 61:12371244

Yoo HD, Gerwick WH (1995) Curacins B and C, new antimi-

totic natural products from the marine cyanobacterium

Lyngbya majuscula. J Nat Prod 58:19611965Zainuddin EN, Mentel R, Wray V, Jansen R, Nimtz M, Lalk M,

Mundt S (2007) Cyclic depsipeptides, ichthyopeptins A

and B, from Microcystis ichthyoblabe. J Nat Prod70:10841088

Zhang L, Ping X, Yang Z (2004) Determination of microcystin-

LR in surface water using high-performance liquid chro-

matography/tandem electrospray ionization mass detector.

Talanta 62:193200

Zheng W, Chen C, Cheng Q, Wang Y, Chu C (2006) Oral

administration of exopolysaccharide from Aphanothecehalophytica (chroococcales) significantly inhibits influ-enza virus (H1N1)-induced pneumonia in mice. Int Im-

munopharmacol 6:10931099

Zou B, Long K, Ma DW (2005) Total synthesis and cytotoxicity

studies of a cyclic depsipeptide with proposed structure of

palauamide. Org Lett 7:42374240


123

Reproduced with permission of the copyright owner. Further reproduction prohibited withoutpermission.

c.10482_2013_Article_9898.pdfCyanobacteria: potential candidates for drug discoveryAbstractIntroductionAnticancerous compoundsCyanobacteria cyclopeptides as a lead compound for cancer treatmentAntiviral compoundsAntibacterial compoundsCyanobacterial bioactive molecules under clinical trialsMethods used for isolation and detection of novel biomoleculesConclusionAcknowledgmentsReferences

out_35.pdf

Documents

Transcript of out_35.pdf