drug discovery from marine sources prof. dr. basvaraj k. nanjwade m. pharm., ph. d department of...

Drug Discovery from Marine Sources

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

Department of PharmaceuticsKLE University College of Pharmacy

BELGAUM-590010Karnataka, INDIA

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• 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.

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INTRODUCTION

Drug Discovery Process and Productivity

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• 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

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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.

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Why Marine?

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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.

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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.

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Marine Organisms for Drug Discovery

• 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).

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Marine Organisms for Drug Discovery

• 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

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New Tools for Drug Discovery

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

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Prospects for SuccessAn Excellent View

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A Variety of Manipulator Tools

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Unique Work Platforms

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How do we choose dive sites?

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600–800m

Resolution 20-50m

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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.

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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.

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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.

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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

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• 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

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Drug Discovery and Development

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Prospects for Success

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Marine Drug Discovery and Development

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Marine Biodiscovery Centre

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Sources of Chemical Diversity

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Generating Chemical Diversity

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Uses of Chemical Diversity

• Purified extract library

• Pure compund library

• Structurally defined pure compound library

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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

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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)

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Facilities

• Collection

• Biological Screening

• Natural Products Chemistry

• Secondary Testing/Pharmacology

• Production of material for clinical evaluation

• Clinical investigation

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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

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Prospects for SuccessCollection Methods

• Biological Diversity = Chemical Diversity

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Prospects for SuccessCollection Strategies

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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

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Prospects for Success Extracts are Complex Mixtures of Marine Products

• The Traditional Way

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Prospects for SuccessNatural Products Chemistry

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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.

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Anti-tumor agents from marine sources currently licensed for development

Organism Metabolite Location Discoverer Current Status

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

Licensed to PharmaMar S.A.

Sponge:Discodermia dissoluta

Discodermolide Caribbean Gunasekera & Longley, HBOI

Licensed to Novartis.

Sponge:Lissodendoryx sp.

Isohomo-halichondrin B

New Zealand Munro & Blunt;Univ. Canterbury, NZ

Licensed to PharmaMar S.A.

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Anti-tumor agents from marine sources currently licensed for development

Organism Metabolite Location Discoverer Current Status

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.

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Anti-tumor agents from marine sources currently licensed for development

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

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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

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Commercially available marine bioproducts

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THANK YOUE-mail: bknanjwade@yahoo.co.in

Cell No: 0091 9742431000