Drug Discovery from Marine Sources Prof. Dr. Basvaraj K. Nanjwade M. Pharm., Ph. D Department of Pharmaceutics KLE University College of Pharmacy BELGAUM-590010

Drug Discovery from Marine Sources

Prof. Dr. Basvaraj K. Nanjwade M. Pharm., Ph. D

Department of PharmaceuticsKLE University College of Pharmacy

BELGAUM-590010Karnataka, INDIA

15th Nov. 2011 1Poona College of Pharmacy, BVDU, Pune.

• Substances produced by living organisms found in nature have played a critical role in the development of drugs for life-threatening conditions.

• Number of recent breakthroughs have resulted in the development and approval of anticancer drugs derived from marine sources such as coral and sponges.

15th Nov. 2011 Poona College of Pharmacy, BVDU, Pune. 2

INTRODUCTION

Drug Discovery Process and Productivity


• The discovery process begins with isolation of unique microbes and generation of chemical extracts of biologically active molecules produced by the microbes.

• Information on the microorganisms, their sources, culture and extraction conditions, novel chemistries and biological activities have been developed that allow for a concentrated effort on the most productive organisms.

• Primary targets leads to identification of novel, biologically active discovery leads in the selected therapeutic areas of oncology and infectious diseases


Discovery Process and Productivity

• The marine environment provides a broad range of diverse habitats from which novel sources of natural products can be derived.

• Studies from around the world have shown that marine organisms produce a diverse array of metabolites with novel chemical structures and potent biological activities as well as other desirable properties.


Why Marine?


Marine Organisms

• Life has originated from the oceans that cover over 70% of the earth’s surface and contain highly ecological, chemical and biological diversity starting from microorganisms to vertebrates.

• The source of unique chemical compounds, which hold tremendous pharmaceutical potential.

• Sources emphasize on investigation of the marine ecosystem to explore numerous complex and novel chemical entities.

• The sources of new leads for treatment of many diseases such as cancer, AIDS, inflammatory conditions, and a large variety of viral, bacterial and fungal diseases.


Marine Organisms for Drug Discovery

• Majority of the marine natural products have been isolated from

• Sponges,

• Coelenterates (sea whips, sea fans and soft corals),

• Tunicates,

• Opisthobranch molluscs (nudibranchs, sea hares, etc.),

• Echinoderms (starfish, sea cucumbers, etc.) and

• Bryozoans (moss animals) and a wide variety of marine microorganisms in their tissues.



• Sponges, the most primitive multicellular invertebrates, considered as a gold mine during the past 50 years, have fascinated scientists for isolation of promising bioactive compounds for human welfare.

• Interestingly, cytarabine (Cytostar-U) also known as Ara-C, a compound isolated from the Caribbean sponge Cryptotheca crypta currently being used with other anticancer drugs in the treatment of acute myelocytic leukaemia (AML).



• Exploration of new locations, collection of organisms never before sampled, and identification of chemicals with pharmaceutical potential.

• New and improved platforms (such as autonomous, remote, and human occupied underwater vehicles) to take us farther and deeper are in development.

• Tools and sensors that have been developed both for space exploration and for diagnostic medicine


New Tools for Drug Discovery


Prospects for Success Harbor Bracnch Operates the Johnson-Sea- Link class subs Depth capability: 3000 ft


Prospects for SuccessAn Excellent View


A Variety of Manipulator Tools


Unique Work Platforms


How do we choose dive sites?


600–800m

Resolution 20-50m



Sensors

• There are numerous opportunities to borrow scientific techniques common to one discipline and apply them innovatively to other areas and questions.

• Different marine cyanobacteria to help select the most promising species for pharmaceutical drug discovery efforts, and then using a genomics and molecular biological approach to understand how their unique natural products are created at a genetic and biochemical level.


Marine Biotechnology

• The genomics revolution has recently provided an orthogonal approach to the discovery of natural product drug leads in which sequenced genomes are “mined” for biosynthetic pathways.

• While promising, this emerging technology faces its own challenges.


Marine Microbial Drug Leads

• Marine invertebrate animals provide a robust source for the discovery of bioactive small molecules with the potential to treat human diseases.

• Metagenomics affords an efficient and inexpensive method to capture this chemical diversity.

• Metagenomics is providing renewed insights into what chemical diversity means and how it originates in the ocean.


Genomic to Animal Marine

• A primary metabolite is a chemical substance which is required for cells to survive and replicate and is therefore required for survival of the organism

• Examples are:– Proteins– Carbohydrates– Lipids– Nucleic acids

Primary Metabolites


• A secondary metabolite is a chemical compound produced by an organism which is not required for survival of the organism but presumably confers an evolutionary advantage

Secondary Metabolites



Drug Discovery and Development


Prospects for Success


Marine Drug Discovery and Development


Marine Biodiscovery Centre


Sources of Chemical Diversity


Generating Chemical Diversity


Uses of Chemical Diversity

• Purified extract library

• Pure compund library

• Structurally defined pure compound library


Screening

• Structural characterisation of organic compounds and element/isotope-tagged biomolecules

• Spectroscopic expertise:

– Nuclear magnetic resonance

– Liquid chromatography - mass spectrometry

– Inductively coupled plasma - mass spectrometry (ICPMS)

– Elemental Imaging: laser ablation ICPMS

– Gas chromatography - ICPMS


Expertise

• 600 & 400 MHz Nuclear Magnetic Resonance

• High and low resolution LC-MS

• High and low resolution ICP-MS

• Parallel HPLC - ESMS/ICPMS

• Fully equipped microbial culture suite

• Separations/chromatography suite (eg SPE, SEC, HPLC)


Facilities

• Collection

• Biological Screening

• Natural Products Chemistry

• Secondary Testing/Pharmacology

• Production of material for clinical evaluation

• Clinical investigation


Prospects for Success Process of Marine Drug Discovery

• Requires a Multidisciplinary Team• Biologists - Marine Biologist

- Cell Biologists; Immunologists, Virologists

- Pharmacologists/Biochemists

- Molecular Biologists

• Chemists

- Natural Products Chemists - Spectroscopists

- Synthetic/Medicinal Chemists

• Business Professionals - Marketing/Technology Transfer/Patent Attorneys15th Nov. 2011 35Poona College of Pharmacy, BVDU, Pune.

Prospects for SuccessMarine Drug Discovery

• Before you collect-Get the right permits

• International Convention on Biological Diversity

– http://www.biodiv.org/convention/articles.asp

• You must have informed consent of host country

– Permits may be issued at a local or country level

– Work with State Governments for State waters; National marine Fisheries for Federal

– Special Permits for Marine Sanctuaries

• Often a Memorandum of Understanding is negotiated prior to collection

– Participation by Host Nation in Project

– Return of Income/Benefit to Host Country

– Reporting requirements

– Sharing of specimens and information on samples15th Nov. 2011 36Poona College of Pharmacy, BVDU, Pune.

Prospects for SuccessPermits for Collections

1. Wading

2. Scuba

3. Trawling

4. Submersibles


Prospects for SuccessCollection Methods

• Biological Diversity = Chemical Diversity


Prospects for SuccessCollection Strategies


Prospects for SuccessScreening Approach

• Cancer

- Pancreatic Cancer

- Multidrug resistant cancers

- Cancer “Specific” Agents (NCDDG)

• Infectious Disease

- Drug resistant staphylococcus aureus

- Anti-malarial-collaborative UCF, WRAIR

• Neurodegenerative Disease-collaborative

- Alzhemer’s

- Neuroprotection (Stroke)

• Inflammation-collaborative15th Nov. 2011 40Poona College of Pharmacy, BVDU, Pune.

Prospects for SuccessFinding Drug to Treat


Prospects for Success Extracts are Complex Mixtures of Marine Products

• The Traditional Way


Prospects for SuccessNatural Products Chemistry


Prospects for SuccessPurification

Organism Metabolite Location Discoverer Current Status

Bryozoan:Bugula neritina

Bryostatin I Gulf of California

Pettit,Arizona State Univ.

In Phase I/IIclinical trials in US/Europe; NCI sponsored trials.

Sea hare:Dolabella auricularia

Dolastatin 10 Indian Ocean Pettit,Arizona State Univ.

In Phase I/IIclinical trials in US; NCI sponsored trials.

Tunicate:Ecteinascidia turbinata

Ecteinascidin 743

Caribbean Rinehart, Univ. Illinois

Licensed to PharmaMar S.A.In Phase II clinical trials inEurope and in US.


Anti-tumor agents from marine sources currently licensed for development


Tunicate:Aplidium albicans

Dehydro-didemnin B [Aplidine]

Mediterranean Rinehart, Univ Illinois

Licensed to PharmaMar S.A.

Gastropod:Elysia rubefescens

Kahalalide F Hawaii Scheuer, Univ. Hawaii


Sponge:Discodermia dissoluta

Discodermolide Caribbean Gunasekera & Longley, HBOI

Licensed to Novartis.

Sponge:Lissodendoryx sp.

Isohomo-halichondrin B

New Zealand Munro & Blunt;Univ. Canterbury, NZ





Actinomycete:Micromonospora marina

Thiocoraline Mozambique Strait

Canedo, Spain Licensed to PharmaMar S.A.

Tunicate:Didemnum granulatum

Isogranulatimide Atlantic [Brazil]

Andersen & Bjerinck, Univ, British Columbia & Brazil

Licensed to Kinetik, Canada.

Sponge: Jaspis sp

Bengamide Fiji Crews et al. Univ. California, Santa Cruz

Synthetic derivative licensed to Novartis.

Sponge:Cymbastella sp.

HemiasterelinsA & B

Papua New Guinea

Andersen, Univ.British Columbia

Licensed to Wyeth-Ayerst.



Product Application Original Source Method of Production

Ara-A Antiviral drug Marine sponge,Cryptotethya cryta

microbial fermentationof analog

Ara-C Anticancer drug Marine sponge,Cryptotethya cryta

Chemical synthesis of analog

Okadaic acid Molecular probe:phosphatase inhibitor

Dinoflagellate Cell culture

Manoalide Molecular probe:phospholipase A2 inhibitor

Marine sponge,Luffariella variabilis

Wild harvest of sponge


Commercially available marine bioproducts

Product Application Original Source Method of Production

Vent ® DNApolymerase

Polymerase chainreaction enzyme

Deep sea hydrothermalvent bacterium

Recombinant protein

Formulaid ®(Martek Biosciences,Columbia, MD)

Fatty acids used as additivein infant formula nutritionalsupplement

Marine microalga Cell culture

Aequorin Bioluminescentcalcium indicator

Bioluminescent jellyfish,Aequora victoria

Recombinant protein

Green FluorescentProtein (GFP)

Reporter gene Bioluminescent jellyfish,Aequora victoria

Recombinant protein


Commercially available marine bioproducts


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Documents

Drug Discovery from Marine Sources Prof. Dr. Basvaraj K. Nanjwade M. Pharm., Ph. D Department of Pharmaceutics KLE University College of Pharmacy BELGAUM-590010