SPONGES AND THEIR POTENTIAL USES by Ranjith Edirisinghe Presented during the Regional Workshop on Underutilized Fish and Marine Genetic Resources and their Amelioration, 10-12 July 2019, Location: Colombo, Sri Lanka
Sustainability by Design: Assessment Tool for Just Energy Transition Plans
SPONGES AND THEIR POTENTIAL USES by Ranjith Edirisinghe
1. SPONGES AND THEIR POTENTIAL USES
Ranjith Edirisinghe
Department of Physical Sciences
Faculty of Applied Sciences
Rajarata University of Sri Lanka
2. Marine Sponges
• Sponges – 5000 -9000 estimated number of species
(diversity is greater on temperate reefs)
• Belong to Kingdom Animalia, Phylum Porifera
• Mainly 3 classes. Calcarea, Hexactinellida and
Demospongiae
• Multicellular, simple body arrangement without
organised tissues or organs. Perforated body contains
endoskeleton of separate spicules
• The sponges have defensive chemical weapons
(secondary metabolites) for their protection
• Rich in terpenoids, steroids, alkaloids, peptides, poly-
ketides… etc
2
3. • Natural products (secondary metabolites) are a critical resource in
the search for new pharmaceuticals, agrochemicals and other
useful chemicals.
• For example, a majority of the pharmaceuticals currently in clinical
use are natural products or are derived from them.
• Almost all commercially important natural products have been
derived from terrestrial sources (mainly plants and soil
microorganisms).
3
4. Marine reefs are highly crowded and extremely
competitive ecosystems.
There is fierce competition for limited space and
nutrients.
4
5. Most marine invertebrates are soft
bodied, sessile or slow moving and
have no protective armour.
They are expected to depend
heavily on secondary
metabolites that are toxic or
objectionable to their
competitors
Predictably they are a rich
sources of natural products
with diverse and potent
biological activities. 5
6. Functions and Importance
• Provides a multitude of functions on
reefs (food for other organisms, forms
numerous symbiotic associations,
maintains reef health)
• Source of food for many marine
organisms, particularly for several
species utilized in the ornamental fish
industry
• Contains bioactive natural products
(Their full potential is yet to be realized)
• Many species of sponges are highly
colourful and attractive which enhances
the beauty of reefs
7. Examples for pharmacologically important
metabolites of sponges
• Calyculins from Discodermia calyx
A novel antitumor metabolite
• Discodermolide from Discodermia dissolute
Inhibit in vitro proliferation of P388
murine leukemia cells Inhibit the
growth of Candida albicans
7
8. • Latrunculins from Latrunculia magnifica
Disrupt microfilament organization
in cultured cells
• Spongistatins from Spongia sp.
and Spirastrella sp.
Act as antineoplastic agents
8
9. 9
• Sri Lanka has a coastline of nearly 1,600 Km with reef
habitats fairly common in the near-shore coastal
waters.There area is estimated to be nearly 680 Km2.
• The biodiversity of these reefs is largely unrecorded
and represent an mostly unexplored and untapped
resource for the discovery of bio-molecules with
potential applications in medicine.
• Very limited studies conducted on sponges in Sri Lanka.
University of Colombo
National Aquatic Resources Research & Development
Agency (NARA)
Rajarata University
10. • National Aquatic Resources Research & Development Agency (NARA)
A Preliminary Study on the Antibacterial Activity of Marine Sponges from East Coast of Sri Lanka. (2007-2009)
Edirisinghe, E.M.R.K.B., Kariyawasam, M.G.I.U. and Rajasuriya, A. Proceedings of the 25th Anniversary International
Scientific Conference of NARA on Tropical Aquatic Research Towards Sustainable Development, p 70, 15 – 16th Feb.
10
Showed antibacterial activity
against Enterobacter aerogenes,
Staphylococcus aureus, Bacillus
subtilis
• University of Colombo
Two Anti-bacterial Metabolites from a Sri Lankan Sponge. (1987)
J.A. Chandrasiri., L.M.V. Thilakaratne, S.A. Deraniyagala and
Ananda Amarasekara. Annual Scientific Sessions of Sri Lanka
Association for Advancement of Science (SLAAS) – Secondary
metabolite - (known compound)
Research on Marine Sponges in Sri Lanka
11. 11
NARA and Uni. Colombo Joint project (2011-2013)
Screening for antibacterial, antifungal and antioxidant activities of some local marine sponges (2012). Ahmad, S.B.N.
Edirisinghe, E.M.R.K.B. and E. Dilip de Silva (2012). Proceedings of the International Conference on Chemical Sciences
(IChem), page 86, Colombo, Sri Lanka, 20-22, June.
Studies on the biomedical activities of some marine sponges in Sri Lanka. (2012). Ahmad, S.B.N., Edirisinghe, E.M.R.K.B.,
Ginigaddarage, P., Rajasooriya, A. and de Silva, E. D. Proceedings of the Scientific Sessions of the National Aquatic Resources
Research & Development Agency (NARA), Colombo, Sri Lanka, 04th December.
Antimicrobial and Antioxidant activities of some selected Sri Lankan Marine Sponges. (2013). Ahmad, S.B.N. MSc Thesis in
Chemistry. University of Colombo
12. 12
Nearly 100 sponges were collected by scuba
diving and tested for antibacterial, antifungal
and antioxidant activity
15. 15
The structure of the active compound was determined at the Dept. of Chemistry
and Oceanography University of British Columbia, Canada
10-bromofascaplysin - having antitumor activity
• Rajarata University
Computer modeling of new drugs using 10-bromofascaplysin
16. University of Colombo
Extensive work on Haliclona (Soestella) sp and Neopetrosia sp
Taxanomic identification, Immuno pharmacological and Toxicological study of
Haliclona (Soestella) sp
Haliclona (Soestella) sp Axinella sp Phakelia sp Stylissa carteri
16
17. 17
• Taxanomic identification, Immuno pharmacological and Toxicological study of Haliclona (Soes tella)
sp: A novel marine sponge species from Sri Lanka. Gunathilake V K, Bertolino M, Bavestrello G,
Ratnasooriya W D, de Silva D and Udagama P V (2017). 17th world sponge conference Ireland.
• Immunomodulation of some selected cytokines in wistar rats by a crude extract of Haliclona
(Soestella) sp: A Sri Lankan marine demosponge. Gunathilake V K, Ratnasooriya W D and Udagama P
V (2015). Drug Discovery and Therapy World congress. Boston, USA.
• Immuno supressive and anti-cancer activity of a novel Sri Lanka marine sponge, Haliclona (Soestella)
Gunathilake V K and Udagama P V (2016).)sp. 12th Euro Biotechnology Congress. Alicante, Spain
• Free radical scavenging effect of a few selected Sri Lankan marine sponges (Demospongiae).
Gunathilake, VK, Ratnasooriya, WD, Udagama PV(2013). Proceeding of the Annual Reseach Symposium,
University of Colombo, Pp 191
• Immunomodulatory activity of Sri Lanka Demospongiae Marine sponge species (Class
Demospongiea): A Pilot Study. Gunathilake, VK, Ratnasooriya, WD, Udagama PV(2013). Proceedings of
The 69th Annual sessions of the Sri Lanka Association for the Advancement of Science, Pp 89.
• Neopetrosia sp, a marine sponge from Sri Lanka with immunomodulatory activities. Gunathilake VK,
Ratnasooriya WD, De Voogd NJ and Udagama P. (2014). Higher Education for the Twnty First Century
Symposium. Pp107.
• In vivo and in vitro inflammatoryactivity of Neopetrosia sp, a marine sponge crude extract in a rat
model. Gunathilake VK, Ratnasooriya WD, De Voogd NJ and Udagama P. (2014).. Proceedings of the 34th
Annual sessions of Institute of Biology, Sri Lanka. Pp42
18. • Modulation of in vitro phagocytic activity, cell proliferation and cytokine production in the Wistar rat
model by a Sri Lankan Haliclona(Soestella) sp sponge crude extract. Gunathilake V K, Ratnasooriya W D
and Udagama PV (2015). Proceedings of the 35th Annual Sessions of the Institute of Biology,Pp 29
• Anti inflammatory activity of Haliclona (Soestella) sp marine sponge crude extract in murine models:
inhibition of pro inflammatory cytokines and leukocyte migration. Gunathilake V K, Ratnasooriya W D
and Udagama P V (2015).. Proceedings of the Annual Research Symposium , University of Colomb, 2015 Pp
112.
• Acute inflammatory and cancer chemopreventive activity of Haliclona (Soestella) sp crude extract: A
novel Sri Lankan marine sponge species. Gunathilake VK and Udagama P V (2016).. Allergy &
Immunology Society of Sri Lanka, 8th Biennial Scientific Sessions. Pp27.
• V K Gunathilake, PhD Thesis - Molecular characterization, isolation and chemical characterization of
Haliclona (Soestella) sp, a novel Sri Lankan marine sponge species. University of Colombo
• V K Gunathilake, BSc Thesis - In vitro anti-inflammatory activity of a few selected Sri Lankan marine
sponge species. University of Colombo
18
19. 19
Determination of In-vitro Antimicrobial Activity of Selected Marine Sponges Found in Sri Lanka.
(2017). Dilrukshi, M. K. G. B., Hettiarachi., S and Edirisinghe. E.M. R. K. B. 10th International Research
Conference of General Sir John Kotelawala Defence University. Sri Lanka. Pp 29, 3-4 August.
Determination of Antioxidant, Antibacterial and Antifungal Activities of Some Marine Sponges of Sri
Lanka. (2017) Senarathne., E.M.N.S and Edirisinghe, E.M.R.K.B. International Conference on Drug
Discovery & Development (ICDDD 2017), Colombo, Sri Lanka. p 20, 12-13 September
In vitro Determination of Anticancer Activity of Selected Marine Sponges of Sri Lanka. (2018).
Senarathne, E.M.N. S., Karunathilake, K.J.K., Rajapakse, R. P. V. J. and Edirisinghe E.M.R.K.B. Proceedings
of the 8th IUPAC International Conference on Green Chemistry, Bangkok, Thailand, p 263. 09-14 September.
Having Anticancer property Stylissa massa
Agelas sp
Rajarata University
20. ANTICANCER ACTIVITY TESTING
• The testings were conducted at University of Peradeniya
• Cytotoxicity of four samples which showed best results for antioxidant and
antimicrobial assays were determined using MTT Assay
Dilution series was prepared using aqueous sponge extracts of selected samples
Cell plates were prepared. Normal cell line : BHK(Baby hamster kidney)
20
21. Addition of the test extracts
Addition of MTT solution
Addition of solubilisation solution
Measurement of absorbance using Elisa reader
Based on the results of cytotoxicity assay,
Anticancer assay was conducted using
HeLa (Henrietta Lacks) as the cancer cell line
21
24. Computer modeling of new drugs using 10-bromofascaplysin
• Cyclin-dependent kinase type 4 (CDK4) is the responsible protein for the G1/S transition in the cell
cycle and which is misregulated in 60-70% of human cancers.
• The natural compound fascaplysin is a kinase inhibitor with enticing selectivity for CDK4.
• The fascaplysin framework has been used to synthesize a series of selective CDK4 inhibitors though in
most cases selectivity was partially lost in the redesign process.
• 10-Bromofascalysin (10FAS) is a natural product which was isolated from Sri Lankan marine sponge
Computer aided drug design
25. Current research
• National Aquatic Resources Research & Development Agency (NARA)
• Leiden University, Netherlands
• University of Colombo, Sri Lanka
• Uppsala University, Sweden
• Rajarata University of Sri Lanka,
• University of British Colombia, Canada
• Leiden University, Netherlands
• University of Peradeniya, Sri Lanka
• University of Sri Jayawardanapura
25
27. • Sampling
Scuba diving skills, lack of knowledge on sponges and other marine resources
• Taxonomy
Identification of sponges
• Facilities for advanced analysis, structural determination, drug design
• HPLC, NMR…..
27
Challenges
28. • Training in taxonomy
• Carry out surveys to determine their abundance, and preparation of a
distribution map of species
• Development of resource management plan and implementation
• Isolate compounds that are potentially useful for developing drugs
• Development of Laboratory facilities for advance analysis
28
Future work
29. 29
Prof. L.M.V. Thilakaratne
Dr. Patrick Hemalal
Prof. Dilip de Silva
Prof. P.V. Udagama
Prof. W.D. Ratnasooriya
Dr. Varuni Gunathilake
Prof. S.A. Deraniyagala
Prof. A.S. Amarasekara
J.A. Chandrasiri
Shakila Ahmad
Prof. R.J. Andersen
Prof. Nicole de Voogd
Arjan Rajasooriya
Bimalka Dilrukshi
Prof. Sanath Hettiarchchi
Prof. R. P. V. J. Rajapakse
Sathsara Senarathna
Acknowledgement
Deishini Herath
The present study mainly focuses on the analysis of the structure of CDK4 (overall conformation changes and overall stability) and investigates whether the 10FAS can selectively bind to CDK4 and, if so, their mode of interaction. Moreover, these investigations may open a new avenue for the design of novel selective inhibitors for CDK4.