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INTERNATIONAL RESEARCH JOURNAL OF PHARMACY www.irjponline.com ISSN 2230 – 8407

Research Article

PREDICTION OF BIOLOGICAL ACTIVITY SPECTRA FOR SECONDARY METABOLITES FROM MARINE MACROALGAE CAULERPA SPP (CHLOROPHYTA – CAULERPALS)

R. Azhaguraj1*, M.C. John Milton1, J. Ganesh1, G. Justin Zenith kumar1 M. Ramakrishnan2

and Stalin Antony2 1Department of Advanced Zoology and Biotechnology, Loyola College, Chennai-600 034, Tamil Nadu, India

2Division of Bioinformatics, Entomology Research Institute, Loyola College Chennai-600 034, Tamil Nadu, India

Article Received on: 20/03/12 Revised on: 11/05/12 Approved for publication: 22/05/12 *E-Mail: azhaguloyolascholar@gmail.com ABSTRACT This study aims to evaluate the biological activity of Caulerpin β-Sitosterol, Taraxerol and Palmtic acid isolated from the marine macro algae Caulerpa spp. The PASS computer program was used in this study to predict the biological activity profile of the four Phenazine derivates. The results were analyzed to show various biological activities like pharmacological (Kinase inhibitor, Neuroprotector and Antiviral), Effects (Oxidoreductase inhibitor, Acid Phosphatase inhibitor) and toxicological activity (Teratogen) of these compounds. The PASS software is useful for the study of biological activity of secondary metabolites. Key words: Macro algae, Natural Products and Biological Activity. INTRODUCTION Seaweeds are wonder plants of the sea and considered as medicinal food of the 21st century. They are non-vascular cryptogenic plants. It is distributed in various ecosystems like intertidal, shallow waters, deep waters, mangroves, estuaries, and lagoons1,2 . Plants and Animals in the sea produce a great variety of compounds, often unique, that serves to protect against its natural enemies or act as chemical cues for reproduction or used for praying. These chemicals are referred as secondary metabolites or natural products and are not involved in the primary metabolism 3. Seaweeds are known marine algae used as food, animal fodder, and fertilizer4. They are used as medicine for their antimicrobial, antitumor, antihyperchlosterolemic, immunomodulation, immunosuppressive, antiulcer activities2. Marine algae in the order Caulerpales (Chlorophyta) are perhaps the most abundant and widely distributed algae in tropical oceans5,6. Algae in the order Caulerpales possess a siphonous or coenocyte construction consisting of multinucleate tubular filaments, lacking cross walls except that to delineate reproductive structures7. Biological activity is a ensuing of the interaction of a chemical compounds with a biological entity. Nature of biological activity depends on the peculiarities of the compound (structure and physico-chemical properties), biological entity (species, sex, age, etc.), mode of treatment (dose, route, etc). Any biologically active compound has a wide spectrum of effects. Some of them are useful in the treatment for diseases, but the others cause various toxic effects. To analysis the biological and toxic effects, there are many software programs are available on the internet. PASS (Prediction of Activity Spectra for Substances) is one of the main programs to analyze the total complex activity of the compound on a biological entity and biological activity spectrum of the substance8. Generally the chemical compounds different types of biological activity was evaluated using PASS software connected to internet, which estimates the probabilities of 900 types of biological activity on the basis of structural formulae of the compound with an accuracy of 85 %. PASS predictions are based on the analysis of structure-activity relationship (SAR) for the training set of

about 46000 biologically active compounds. Already PASS has been used to predict the anti-HIV activity9, activities of Flavonoid derivatives10 antitumor activity 11. Antimalarial activity12 and activities of essential oils13. In the present study we have described the biological activity of Caulerpin, β-Sitosterol, Taraxerol and Palmtic acid isolated from the Genus Caulerpa spp. MATERIALS AND METHODS The chemical structures of the Phenazine derivatives14,15

Caulerpin (C24H18N2O4), Taraxerol (C30H50O), β-Sitosterol (C29H50O) and Palmtic acid (C16 H32 O2) present in the Caulerpa lamourouxii, Caulerpa sertulariodes, Caulerpa racemosa var clavifera were obtained from Pubchem compound repository (http://www.ncbi.nlm.nih.gov/ pccompound) (Figure 1). The structures were drawn using the Chem sketch package 11.0 belonging to the ACD chem. laboratory9,11,13,16. The biological activity spectrum was predicated by PASS 8, 17,18. RESULTS AND DISCUSSION Four Phenazine derivatives namely caulerpin, taraxerol, β-sitosterol and Palmtic acid were analyzed by the PASS for their different types of biological activity. Pa (probability to be active) and Pi (probability to be inactive) were estimates of probability for the compound to be active and inactive respectively for each type of activity from the biological activity spectrum. Their values vary from 0.000 to1.000. The results showed that caulerpin (Table1) could possess biological activities like ulcerogenic and neuroprotector 9,11,13,16. Taraxerol (Table 2) and β-Sitosterol (Table 3) exhibited similar effect and inhibitor mechanism like antiviral, hepatoproctent, apoptosis, agonist and antifertility (female). Palmtic acid (Table 4) possesses the activities like hematotoxic, antiviral (Arobovirus), antithrombotic, antimutagenic, urease inhibitor. The above predicated results shows the available information on the pharmacological and toxicologal activity/mechanism /effects of these compounds, and were corroborative with the previous reports 9-13,16,19 A large number and variety of herbivore, ranging from highly mobile macro grazers (mammals, fish, sea urchin and large crustaceans and gastropods) to smaller more sedentary meso-grazers (small gastropods, amphipods, isopods and

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polychaetes) consume equally large numbers and variety of marine macro algae (green/red/brown) seaweeds and produce a variety of different secondary metabolites that serve to protect against the pathogens, grazers and phytal fauna20. Marine algae Caulerpa spp are defended from herbivory by a variety of secondary metabolites including polyphenols, acetogenins, terpens, amino and halogenated compounds which can influence palatability and digestibility of the host algae21. These metabolites deterred feeding by marine herbivores in the field and laboratory assay22. Chemical defenses indirectly structure algal populations and communities in coral reef and other near shore habitats. Algae also produce higher concentrations of defensive compounds in areas subject to high herbivore pressure7. The value of marine algal secondary metabolites has shown that many of these compounds function as chemical weapons and have evolved into highly potent inhibitors of physiological process in the prey, predators or competitors of the marine organisms that use them. PASS results shows that the pharmacological and toxicological activity of these compounds are different but this difference may be due to the diversity among target sites and the reciprocal activity of the specific molecules towards such diversified targets and their differential physiological activity. The application of computerized system PASS could be used in toxicoinformatics and chemical ecology. The results shows that secondary metabolites isolated from the marine macro algae Caulerpa spp could possess several pharmacological activities such as ulcerogenic, hyper thermic, anti-neoplastic, antioxidant, antipruritic, hypercholesterolemia, neurotoxic, anti-inflammatory, antiviral (Arbovirus), apoptosis agonists, antithrombotic, nephrotoxic, oxidoreductase inhibitor, kinase inhibitor, phosphatase inhibitor, and TERT expression inhibitor. ACKNOWLEDGMENT We thank Mr. Nagoor, ERI, Loyola College, Chennai for his early revision and correction of this manuscript. REFERENCES 1. Sakthivel. Seaweed Cultivation – A profitable venture for the Economic

Rehabilitation of coastal Poor .In Kannaiyan .S. and Venkataraman.(eds). Biodiversity Conservation in Gulf of Manner Biosphere Reserve. National Biodiversity Authority, Chennai. 2007; 484pp.

2. Anatharaman. P, G. Thirumaran and T. Balasubramanian. Seaweed Farming-Alternative Livelihood. In Kannaiyan .S. and Venkataraman. (Eds). Biodiversity Conservation in Gulf of Manner Biosphere Reserve. National Biodiversity Authority, Chennai. 2007; 484pp.

3. Canneli R.J.P. How to approach the isolation of a natural product .In Canneli R.P.J (Ed.) Natural Products Isolation, Methods in Biotechnology. Volume 4, Humana Press, Totowa, New Jersey.1998.

4. Smith, G.M. Marine Algae of the Monterey Peninsula, California. Stanford Univ., 2nd Edition. 1944; 275pp.

5. Hillis Colinnvaux.L. Ecology and taxonomy of Halimeda. Primary producer of coral reefs.Adv.Mar.Biol.1980; 17:1-327.

6. Norris J.N and Fenical W. Chemical defense in tropical marine algae. In Rutzler K. Macintyre I. G (Eds).The Atlantic barrier reef ecosystem at Carne Bow Cay Bellizer. Structure and communities. Smithsonian Contr. Mar. Sci.1982; 12:417-431.

7. Paul, V J., Fenical, W. Chemical defense in tropical Green algae, order Caulerpales. Mar. Ecol. Prog. Ser. 1986; 34: 157-169.http://www.int-res.com/articles/meps/34/m034p157.pdf

8. Porovikov V., Filimonov D., Borodina, Yu., Lagunin, A and Kos, A. Robustness of Biological Activity Spectra Predicting by Computer Program PASS for Non-Congeneric Sets of Chemical Compounds. J. Chem. Inform. Comput. Sci. 2000;40:1349-1355. http://www.akossamples.com/pass/articles/J.%20Chem. %20 Inf.%20 Comput. % 20Sci .%2040%202000.pdf

9. Maridass M, Raju G, Thangavel K and Ghanthikumar S. Predication of anti-HIV activity of flavonoid constituents through PASS.

Ethnobotanical leaflets, 2008; 12: 954-94. http:/ /opensiuc.lib.siu.edu/cgi/viewcontent.cgi? article=1165 &context=ebl

10. Maridass M. PASS: Prediction of activity spectra for biologically active constituent of Polygodial (A Drimane type of Dialdehyde Sesquiterpene) Indian Journal of Multidisciplinary Research. 2008; 3:191-197.

11. Azhaguraj R., E. Arokia Lenin, C. Viswanathan, B. Sangeetha and M. Selvanayagam Prediction of Biological Activity of Algal Antitumor Drugs Using PASS. Pharmacologyonline, 2010;3:22-34. http://pharmacologyonline.silae.it/files/archives/2010/vol3/03.Lenin.pdf

12. John de Britto .Prediction of biological activity of anti-malarial drugs using PASS. Indian Journal of Multidisciplinary Research. 2008; 4(2): 271-274.

13. Abiya Chelliah D. Biological activity predication of an ethno medicinal plant Cinnamomum camphora through Bio-informatics. Ethnobotanical leaflets 2008; 12: 181-190. http://opensiuc.lib.siu.edu/cgi/ viewcontent.cgi?article=1056&context=ebl

14. Santos, A. G.; Doty, M. S. Lloydia 1971; 34: 88. 15. Bhakuni D.S., D.S. Rawat Bioactive Marine Natural Products, Anamaya

Publishers, New Delhi, India. 2005; 1-39 16. John de Britto, Leon Stephan Raj T, and Abiya Chelliah D. Prediction of

biological activity spectra for few anticancer drugs derived from plant sources. Ethnobotanical leaflets, 2008;12: 801-810. http:// opensiuc.lib.siu.edu /cgi/viewcontent.cgi?article=1144 & context=ebl

17. Alexy Lagunin, Alla Stepanchikova, Dmitrii Filimonov and Vladimir Poroikov. PASS: Prediction of activity spectra for biologically active substances.Bioinformatics.2000;16(8):747-748. http://bioinformatics.oxfordjournals.org/content/16/8/747.full.pdf+html

18. Sadym A, Lagunin A, Filimov D and Poroikov V. Predication of biological activity spectra via the internet. SAR and QASR in Environmental Research 2003,14(5-6):339-347.http:/ /www.tandfonline.com DOI 10.1080/10629360310001623935

19. Ayyad S-E-N, Abdal-Halim OB, Shier WT and Hoye TR. Cytotoxic hydroazulene diterpens from the brown algae Cystoseria myrica. Z. Biosci. 2003; 58: 33-38.

20. Hay M.E and Steinberg P.D. The chemical ecology of plant herbivore interactions in Marine verses terrestrial communities. In G.A. Rosenthal and Berenbaum (Eds).Herbivores: Their interactions with plant secondary metabolites. Academic Press. New York. 1992; 371-413.

21. Paul V.J.E. Cruz Rivera and R.W. Thacker. Chemical mediation of macro algae- herbivore interactions. Ecological and Evolutionary Perspectives. In. Mcclintock J.B and B.J.Baker (eds).Marine Chemical Ecology.CRC Press. Boca Raton/Florida. 2001; 22-265.

22. Hay, M. E., Duffy, J. E., Fenical, W. (1990). Host-plant specialization decreases predation on a marine amphipod: an herbivore in plant's clothing. Ecology. 7(2): 733-743. http:// smartech.gatech.edu/bitstream /handle /1853/42185/ Hay_Ecology_1990_001 .pdf? sequence=1.

Table 1. Biological activity of Caulerpin (C24H18N2O4).

Pa Pi Activity

0.773 0.020 Amyotrophic lateral sclerosis treatment

0.748 0.005 Ulcerogenic

0.721 0.009 Kinase inhibitor

0.719 0.029 Protoporphyrinogen oxidase inhibitor

0.719 0.033 Neuroprotector

Table 2. Biological activity of Taraxerol (C30H50O). Pa Pi Activity

0.937 0.005 Mucomembranous protector 0.920 0.005 Phosphatase inhibitor 0.854 0.003 Antiviral (Influenza) 0.848 0.004 Oxidoreductase inhibitor 0.804 0.004 Phospholipase C inhibitor 0.806 0.012 Membrane integrity antagonist 0.785 0.005 Hepatoprotectant 0.786 0.011 Hypercholesterolemic 0.744 0.003 DNA ligase (ATP) inhibitor 0.729 0.006 Antinephritic 0.713 0.004 Antiinfertility, female 0.722 0.021 Apoptosis agonist 0.711 0.016 Antineoplastic 0.705 0.027 Opioid dependency treatment 0.706 0.038 Cholesterol synthesis inhibitor

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Table 3. Biological activity of β- Sitosterol (C29H50 O).

Pa Pi Activity Pa Pi Activity

0.959 0.001 DELTA14-sterol reductase inhibitor 0.788 0.036 Acylglycerol lipase inhibitor

0.957 0.002 Cholesterol antagonist 0.752 0.002 Cholestenone5alpha-reductase inhibitor

0.958 0.003 Antihypercholesterolemic 0.753 0.004 Peptidoglycan glycosyltransferase inhibitor

0.917 0.005 Prostaglandin-E2 9-reductase inhibitor 0.749 0.004

Trans-octaprenyltranstransferase

inhibitor

0.848 0.004 Oxidoreductase inhibitor 0.749 0.009 1-Acylglycerol-3-phosphate O-acyltransferase inhibitor

0.844 0.016 Teratogen 0.753 0.021 Linoleate diol synthase inhibitor

0.824 0.005 Hypercholesterolemic 0.752 0.020 Nephrotoxic

0.808 0.005 Alkenylglycerophosphoethanolamine hydrolase inhibitor 0.736 0.006 Trans-1,2-dihydrobenzene-1,2-

diol dehydrogenase inhibitor 0.829 0.029 Phosphatase inhibitor 0.734 0.005 Nerve growth factor agonist 0.800 0.004 Hepatoprotectant 0.731 0.004 Antiinfertility, female

0.798 0.012 Apoptosis agonist 0.725 0.008 Alcohol O-acetyltransferase inhibitor

0.798 0.014 Cholesterol synthesis inhibitor 0.717 0.004 Signal peptidase I inhibitor

0.796 0.017 Squalene-hopene cyclase inhibitor 0.714 0.003 Cycloartenol synthase inhibitor

0.777 0.001 Cholesterol oxidase inhibitor 0.709 0.006 Phospholipase C inhibitor

0.780 0.004 N-(long-chain-

acyl)ethanolamine deacylase inhibitor

0.728 0.035 Toxic

0.791 0.018 Dextranase inhibitor 0.718 0.027 Glucan endo-1,3-beta-D-glucosidase inhibitor

0.779 0.008 Cholestanetriol 26-monooxygenase inhibitor 0.747 0.059 Transcription factor inhibitor

0.765 0.006 Atherosclerosis treatment 0.704 0.112 Mucomembranous protector

Table 4. Biological activity of Palmtic acid (C16 H32 O2). Pa Pi Activity Pa Pi Activity

0.950 0.002 Dextranase inhibitor 0.912 0.016 Methylenetetrahydrofolate

reductase (NADPH) inhibitor

0.942 0.003 Gaucher disease treatment 0.899 0.005 Superoxide dismutase inhibitor

0.944 0.006 Hematotoxic 0.893 0.003 Hydroxylamine reductase (NADH) inhibitor

0.940 0.005 Acylglycerol lipase inhibitor 0.890 0.002 Skin diseases treatment 0.937 0.003 Sarcosine oxidase inhibitor 0.892 0.004 Lipoprotein lipase inhibitor

0.936 0.002 Phosphatidylglycerophosphatase inhibitor 0.891 0.004 All-trans-retinyl-palmitate

hydrolase inhibitor

0.936 0.003 Glucan endo-1,3-beta-D-glucosidase inhibitor 0.899 0.011 Benzoate-CoA ligase

inhibitor

0.935 0.003 Pullulanase inhibitor 0.890 0.004 IgA-specific

metalloendopeptidase inhibitor

0.932 0.005 Pro-opiomelanocortin converting enzyme inhibitor 0.887 0.003 Glucan 1,4-alpha-

maltotriohydrolase inhibitor

0.928 0.003 Poly(alpha-L-guluronate) lyase inhibitor 0.885 0.003 Aminoacylase inhibitor

0.926 0.003 Exoribonuclease II inhibitor 0.894 0.012 Beta-adrenergic-receptor kinase inhibitor

0.925 0.002 Poly(beta-D-mannuronate) lyase inhibitor 0.885 0.004 Aspartate-ammonia ligase

inhibitor

0.925 0.002 Xylan endo-1,3-beta-xylosidase inhibitor 0.875 0.003 Succinate-CoA ligase

(ADP-forming) inhibitor

0.925 0.002 Glutarate-semialdehyde dehydrogenase inhibitor 0.870 0.003 (S)-2-hydroxy-acid oxidase

inhibitor

0.925 0.003 D-lactaldehyde dehydrogenase inhibitor 0.867 0.003 Alkenylglycerophosphoethanolamine hydrolase inhibitor

0.923 0.003 Methylamine-glutamate N-methyltransferase inhibitor 0.870 0.006 Cutinase inhibitor

0.922 0.004 Levanase inhibitor 0.865 0.003 Leukotriene-B4 20-monooxygenase inhibitor

0.916 0.003 NADPH oxidase inhibitor 0.866 0.006 Pulmonary hypertension treatment

0.918 0.007 Mucomembranous protector 0.861 0.004 N-formylmethionyl-peptidase inhibitor

0.912 0.002 Prostaglandin-A1 DELTA-isomerase inhibitor 0.862 0.007 Creatininase inhibitor

0.912 0.005 Fucosterol-epoxide lyase inhibitor 0.856 0.003 Amidase inhibitor

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0.910 0.004 L-glucuronate reductase inhibitor 0.855 0.003 Creatinase inhibitor

0.908 0.005 Antiseborrheic 0.856 0.004 Rhamnulose-1-phosphate aldolase inhibitor

0.905 0.003 Peptide-N4-(N-acetyl-beta-

glucosaminyl)asparagine amidase inhibitor

0.858 0.008 Linoleate diol synthase inhibitor

0.902 0.003 Gluconate 5-dehydrogenase inhibitor 0.853 0.004 Phenol O-methyltransferase inhibitor

0.900 0.002 Pectin lyase inhibitor 0.852 0.002 N-(long-chain-

acyl)ethanolamine deacylase inhibitor

0.853 0.004 Alanine transaminase inhibitor 0.835 0.005 NADH kinase inhibitor

0.854 0.008 Sickle-cell anemia treatment 0.834 0.004 Carboxypeptidase B inhibitor

0.860 0.014 Protein-glutamate methylesterase inhibitor 0.839 0.010 Beta-mannosidase inhibitor

0.847 0.004 Aldehyde dehydrogenase (NADP+) inhibitor 0.831 0.004

2-Haloacid dehalogenase (configuration-inverting)

inhibitor

0.847 0.005 3-Phytase inhibitor 0.828 0.003 Carboxypeptidase U inhibitor

0.844 0.002 Plasmanylethanolamine desaturase inhibitor 0.827 0.003 Sclerosant

0.848 0.007 IgA-specific serine endopeptidase inhibitor 0.826 0.003 L-lysine 6-transaminase

inhibitor

0.848 0.008 Cl--transporting ATPase inhibitor 0.838 0.016 Peptidyl-dipeptidase Dcp inhibitor

0.843 0.003 Catalase inhibitor 0.835 0.014 Monodehydroascorbate reductase (NADH) inhibitor

0.850 0.012 Arylsulfate sulfotransferase inhibitor 0.830 0.009 Anthranilate-CoA ligase inhibitor

0.841 0.004 2-Oxoglutarate decarboxylase inhibitor 0.825 0.004 Galactolipase inhibitor 0.840 0.003 Thiosulfate sulfurtransferase inhibitor 0.824 0.004 Leukopoiesis stimulant 0.841 0.004 NMDA receptor glycine site agonist 0.825 0.005 Candidapepsin inhibitor

0.841 0.005 Lysostaphin inhibitor 0.824 0.004 Stearoyl-CoA 9-desaturase inhibitor

0.838 0.002 Guanidinobutyrase inhibitor 0.830 0.011 Glutamine-phenylpyruvate transaminase inhibitor

0.839 0.004 Licheninase inhibitor 0.822 0.005 Cyclomaltodextrinase inhibitor

0.845 0.010 Laccase inhibitor 0.823 0.005 Urethanase inhibitor

Source of support: Nil, Conflict of interest: None Declared