on Marine Natural Products - MANAPRO 2013

Cubierta 4:Abstracts 01/08/13 20:01 Página 1

on Marine Natural Products


14th International Symposium 8th European Conference


September 15th - 20th, 2013 La Toja Island, Galicia Organized by:

Spain

Programme and AbstractSPART

Interior ABSTRACTS:Abstracts 02/08/13 11:48 Página 1


on Marine Natural Products September 15th - 20th, 2013 La Toja Island, Galicia Spain

Organized by:


Committees

Committees

Honor Committee

President H.R.H. the Prince of Asturias

Members Mr. Alberto Núñez Feijóo, President of the Xunta de Galicia Mr. Luis de Guindos, Minister of Economy and Competitiveness Mr. Miguel Ángel Pérez García, Mayor of O´Grove Prof. José María Fernández-Sousa, President of Zeltia Group

International Advisory Board Committee Prof. William Fenical, University of California, San Diego, USA Prof. Ian Paterson, University of Cambridge, UK Prof. Manuel Norte, University of La Laguna, Spain Prof. Heonjoon Kang, Seoul National University, South Korea Prof. Marcel Jaspars, University of Aberdeen, UK Prof. Raymond Andersen, University of British Columbia, Canada

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14th International Symposium on Marine Natural Products / 8th European Conference on Marine Natural Products


Commitees

Committees

Organising Committee Dr. Carmen Cuevas, Chair, R&D, PharmaMar, Spain Prof. Vassilios Roussis, Secretary, University of Athens, Greece Prof. Carlos Jiménez, Secretary, University of La Coruña, Spain

Local Organising Committee Prof. Fernando Albericio, University of Barcelona, Spain Prof. Ricardo Riguera, University of Santiago de Compostela, Spain Dr. Simon Munt, R&D, PharmaMar, Spain Dr. Marta Pérez, R&D, PharmaMar, Spain Dr. Andrés Francesch, R&D, PharmaMar, Spain Dr. Fernando de la Calle, R&D, PharmaMar, Spain Mr. Santiago Bueno, R&D, PharmaMar, Spain Ms. Carmen Eibe, Zeltia Group, Spain Mr. Juan Manuel Báez, Zeltia Group, Spain Ms. Olga Chousa, R&D, PharmaMar, Spain


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Conference Sponsors

Conference Sponsors

Platinum

Gold

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Conference Sponsors

Conference Sponsors

Silver

Bronze


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Programme at a glance Headquarters hotel for the conference: Hotel Hesperia La Toja SUNDAY 15th September, 2013

Programme at a glance

20:00 Welcome Reception MONDAY 16th September, 2013

Marine Natural Products: Celebrating Success 09:00 - 09:05 Carmen Cuevas, Chair of the Congress: Welcome 09:05 - 10:00 OPENING CEREMONY. Invited Authorities MORNING SESSION: Marketed Marine Natural Products: Success stories Chair: Murray Munro Eisai, USA 10:05 - 10:50 L 1 Bruce A. Littlefield 10:50 - 11:20 Coffee break PharmaMar, Spain 11:20 - 12:05 L 2 José Mª Fernández Sousa-Faro Marinomed, Austria 12:05 - 12:50 L 3 Andreas Grassauer Bioalvo, Portugal 12:50 - 13:35 L 4 Ana Martins 13:35 - 15:00 Lunch

Supply Challenge: Marine Natural Products as potential therapeutic agents AFTERNOON SESSION: Marine Natural Products in Development Chair: Ian Paterson 15:00 - 15:40 PL 1 William Fenical 15:40 - 16:10 IL 1 Raymond Andersen 16:10 - 16:40 IL 2 Simon Munt 16:40 - 17:00 Coffee break 17:00 - 17:20 OC 1 Julian Adams 17:20 - 17:40 OC 2 Laurent Meijer 17:40 - 18:00 OC 3 Heonjoong Kang 18:00 - 18:15 Fernando Albericio POSTER SESSION 1 - Hotel Hesperia 18:30 - 21:30 Posters 20:00 Refreshments

University of California, San Diego, USA University of British Columbia, Canada PharmaMar, Spain Infinity Pharmaceuticals, USA Manros Therapeutics, France Seoul National University, South Korea Marine Drugs. e-conference platform Sciforum

TUESDAY 17th September, 2013

Supply Challenge: Marine Natural Products as potential therapeutic agents MORNING SESSION: Chemical Synthesis Chair: Tadeusz Molinski 09:00 - 09:40 PL 2 Ian Paterson 09:40 - 10:10 IL 3 Jef K. De Brabander 10:10 - 10:30 OC 4 Brent Copp 10:30 - 10:50 OC 5 Thomas Lindel 10:50 - 11:30 Coffee break 11:30 - 11:50 OC 6 Nicolas Inguimbert 11:50 - 12:10 OC 7 Marta Pelay Gimeno 12:10 - 12:30 OC 8 Daniel Romo 12:30 - 13:00 IL 4 Angelo Fontana 13:00 - 14:30 Lunch AFTERNOON SESSION: Biosynthesis: the "omics" era Chair: Vassilios Roussis 14:30 - 15:10 PL 3 Bradley Moore 15:10 - 15:40 IL 5 Jörn Piel 15:40 - 16:00 OC 9 Letícia V. Costa-Lotufo 16:00 - 16:20 OC 10 Brandon I. Morinaka 16:20 - 17:00 Coffee break 17:00 - 17:20 OC 11 Toshiyuki Wakimoto 17:20 - 17:40 OC 12 Nadine Ziemert 17:40 - 18:10 IL 6 Pei-Yuan Qian POSTER SESSION 2 - Hotel Hesperia 18:30 - 21:30 Posters 20:00 Refreshments

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University of Cambridge, UK UT Southwestern Medical Center, USA University of Auckland, New Zealand TU Braunschweig, Germany University of Perpignan, France Institute for Research in Biomedicine, Spain Texas A&M University, USA CNR - Istituto di Chimica Biomolecolare, Italy

Scripps Institution of Oceanography, USA Eidgenössische Technische Hochschule Zurich, Switzerland Universidade Federal do Ceara, Brasil Eidgenössische Technische Hochschule Zurich, Switzerland The University of Tokyo, Japan Scripps Institution of Oceanography, USA Hong Kong University of Science & Technology, China



Programme at a glance Headquarters hotel for the conference: Hotel Hesperia La Toja WEDNESDAY 18th September, 2013

Programme at a glance

Social Activities THURSDAY 19th September, 2013

Unlocking the Potential of Marine Organisms MORNING SESSION: Frontiers of Discovery & Innovation Chair: Junichi Tanaka 09:00 - 09:40 PL 4 Tadeusz Molinski University of California, San Diego, USA 09:40 - 10:10 IL 7 M. Valeria D'Auria University of Naples "Federico II", Italy 10:10 - 10:30 OC 13 Ronald J. Quinn Eskitis Institute, Griffith University, Australia 10:30 - 10:50 OC 14 Mark Hamann University of Mississippi, USA 10:50 - 11:30 Coffee break 11:30 - 11:50 OC 15 Matthias Köck Alfred-Wegener-Institut, Germany 11:50 - 12:10 OC 16 Gabriele Koenig University Bonn, Germany 12:10 - 12:30 OC 17 William H. Gerwick University of California San Diego, USA 12:30 - 12:50 OC 18 Philipp Hess Ifremer, Laboratoire Phycotoxines, France 12:50 - 13:20 IL 8 Shigeki Matsunaga The University of Tokyo, Japan 13:20 - 14:30 Lunch AFTERNOON SESSION: Marine Ecology: Understanding Marine Organisms Chair: Raymond Andersen 14:30 - 15:10 PL 5 Patrizia Ciminiello University of Naples "Federico II", Italy 15:10 - 15:40 IL 9 Vassilios Roussis University of Athens, Greece 15:40 - 16:00 OC 19 Marie-Lise Bourguet-Kondracki UMR 7245 CNRS-MNHN, France 16:00 - 16:20 OC 20 Miguel Costa Leal Universidade de Aveiro, Portugal 16:20 - 17:00 Coffee break Special Symposium for Young Investigators Chair: William Fenical 17:00 - 17:40 PL 6 Brian Murphy 17:40 - 18:10 IL 10 Gordon Florence 20:00 Gala Dinner

Illinois University, USA University of St. Andrews, UK

FRIDAY 20th September, 2013

Special Symposium for Young Investigators Chair: William Fenical 09:00 - 09:30 IL 11 09:30 - 09:50 OC 21 09:50 - 10:10 OC 22 10:10 - 10:30 OC 23 10:30 - 10:50 OC 24 10:50 - 11:30 11:30 - 11:50 OC 25 11:50 - 12:15 12:15 - 12:30 12:30 - 12:45 12:45 - 13:15 13:15 - 14:30

Antonio Hernández Robert Britton Robert Brkljaca Efstathia Ioannou Kine Østnes Hanssen Coffee break Paula Jiménez Closing speech: International Advisory Board Committee Closing speech: Chair of the Congress Closing speech: President of PharmaMar Selection of the host for the 15th MaNaPro Symposium Lunch

University of La Laguna, Spain Simon Fraser University, Canada Rmit University, Australia University of Athens, Greece University of Tromsø, Norway Labomar - Universidade Federal do Ceara, Brazil

L: Lecture (45') PL: Plenary Lecture (40') OC: Oral Communication (20') IL: Invited Lecture (30')


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Listed Programme MONDAY 16th September, 2013

Marine Natural Products: Celebrating Success

Listed Programme

MORNING SESSION: Marketed Marine Natural Products: Success stories L1

Bruce A. Littlefield Eisai, USA

L2 L3 L4

José Mª Fernández PharmaMar, Spain Andreas Grassauer Marinomed, Austria Ana Martins Bioalvo, Portugal

Development of Eribulin, A Macrocyclic Ketone Analog of Halichondrin B, for Advanced Breast Cancer Discovery and Development of trabectedin Antiviral Marine Biopolymers - From concepts to the market A new Botulin-like Neuromodulator Bioactive derived from a deep sea Microorganism: Applications for daily skin care

Supply Challenge: Marine Natural Products as potential therapeutic agents AFTERNOON SESSION: Marine Natural Products in Development PL 1 William Fenical IL 1

Raymond Andersen IL 2 Simon Munt OC 1 Julian Adams OC 2 Laurent Meijer

University of California, San Diego, USA University of British Columbia, Canada PharmaMar, Spain Infinity Pharmaceuticals, USA Manros Therapeutics, France

OC 3 Heonjoong Kang

Seoul National University, South Korea Fernando Albericio Institute for Research in Biomedicine, IRB Barcelona, Spain

Seriniquinone, a Marine Microbial Metabolite for the Treatment of Melanoma Discovery and Synthesis of Marine Natural Product Drug Leads Isolation, synthesis, preclinical and clinical development of PM060184 HSP90 Inhibitors in the clinic: Retaspimycin HCl (IPI-504) in non-small cell lung cancer (NSCLC) Leucettines, a family kinase inhibitors derived from the Marine Sponge Alkaloid Leucettamine B. Development as drug candidate for down syndrome and Alzheimer's disease Potential drug leads for diabetes, osteoporosis and bacterial infection from Marine Organisms Special issue "Selected Papers from the 14th International Symposium on Marine Natural Products" Journal Marine Drugs. e-conference platform Sciforum (www.sciforum.net)

TUESDAY 17th September, 2013

Supply Challenge: Marine Natural Products as potential therapeutic agents MORNING SESSION: Chemical Synthesis PL 2 Ian Paterson IL 3 Jef K. De Brabander OC 4 Brent Copp

University of Cambridge, UK UT Southwestern Medical Center, USA University of Auckland, New Zealand OC 5 Thomas Lindel TU Braunschweig, Germany OC 6 Nicolas Inguimbert University of Perpignan, France OC 7 Marta Pelay Institute for Research in Gimeno Biomedicine, Spain OC 8 Daniel Romo Texas A&M University, USA IL 4

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Angelo Fontana

Istituto di Chimica Biomolecolare, CNR, Italy

Challenges and Discoveries in Marine Macrolide Synthesis Natural Products: an opportunity for discovery research The discovery and development of new antimalarial drug leads from libraries of ascidian natural products Novel access to Cubitane- and Eunicellane-Type Diterpenoids Revision of the Laxaphycin B structure based on its synthesis and possible generalisation to its analogs First total synthesis of the antitumoral Marine Cyclodepsipeptide Pipecolidepsin A Innovations in Synthetic Efficiency and Natural Product Derivatization Driven by Natural Products and Impacting Cell Biology Bioactive Metabolites from Marine Protists



Listed Programme AFTERNOON SESSION: Biosynthesis: the "omics" era Bradley Moore

IL 5

Jörn Piel

OC 9

Letícia V. Costa-Lotufo OC 10 Brandon I. Morinaka OC 11 Toshiyuki Wakimoto OC 12 Nadine Ziemert IL 6

Pei-Yuan Qian

Scripps Institution of Oceanography, USA Eidgenössische Technische Hochschule (ETH) Zurich, Switzerland Universidade Federal do Ceara, Brasil Eidgenössische Technische Hochschule (ETH) Zurich, Switzerland The University of Tokyo, Japan Scripps Institution of Oceanography, USA Hong Kong University of Science & Technology, China

Marine bacterial biosynthesis of Polybrominated Organic Compounds Single-cell-GUIDED identification of natural product sources in complex sponge microbiomes Metabolomics Analysis of Zoanthids collected along the Brazilian Coast suggests a correlation between Microbial content and Cytotoxicity Using the Polytheonamide Pathway to Extend the Structural Diversity of Ribosomal peptides A Bacterial Symbiont of Japanese's Marine Sponge Discodermia Calyx, Produces Biologically active Metabolites Genomic insights into the diversity and evolution of secondary metabolite genes in the Marine Actinomycete Genus Salinispora Didemnins and Thalassospiramides—Gifts awarded by Genome mining of Tistrella

THURSDAY 19th September, 2013

Unlocking the Potential of Marine Organisms MORNING SESSION: Frontiers of Discovery & Innovation Tadeusz Molinski University of California, San Diego, USA IL 7 Maria Valeria Università di Napoli, Italy D’Auria OC 13 Ronald J. Quinn Eskitis Institute, Griffith University, Australia OC 14 Mark T. Hamann University of Mississippi, USA PL 4

OC 15 Matthias Köck

Alfred-Wegener-Institut, Germany

OC 16 Gabriele Koenig

University Bonn, Germany

OC 17 William H. Gerwick OC 18 Philipp Hess

University of California, San Diego, USA Ifremer, Laboratoire Phycotoxines, France University of Tokyo, Japan

IL 8

Shigeki Matsunaga

Heterocyclic Compounds from Marine Sponges: Synthesis in Aid of Structure Elucidation and SAR Marine Compounds as Modulators of metabolic nuclear receptors: Isolation, Design, Total Synthesis and potential Therapeutic Application Computational Methods in Marine Natural Products and fingerprinting the drug-like natural product metabolome Using Host-Associated Bacteria and Synthesis to construct complex and novel Marine Cytotoxins with unique Mechanisms of Action. New Cystochromanes and oxidized Meroditerpenes from the Brown Alga Cystoseira Baccata. Indication for a general revision of the Bicyclo[4.3.0]Nonane System? Salimabromide: Unexpected Chemistry from the obligate Marine Myxobacterium Enhygromyxa Salina Bastimolide A, A Biosynthetically intriguing Polyhydroxy Macrolide with Potent Antimalarial Activity High resolution mass spectrometry and biological tools for screening of marine biotoxins and discovery of marine natural products via dereplication Marine natural products with anti-cancer related activities


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Listed Programme

PL 3


Listed Programme THURSDAY 19th September, 2013

Listed Programme

Unlocking the Potential of Marine Organisms MORNING SESSION: Frontiers of Discovery & Innovation PL 5 Patrizia Ciminiello University of Napoli Mediterranean Harmful Algal Bloom Events: Old and New Risks "Federico II", Italy IL 9 Vassilios Roussis University of Athens, Greece Chemical Ecology for Ecological Chemicals OC 19 Marie-Lise UMR 7245 CNRS-MNHN, Exploring the Microbial diversity associated with a Carnivorous Sponge Bourguet-Kondracki France OC 20 Miguel Costa Leal Universidade de Aveiro, Biogeography in Marine Natural Products Research Portugal Special Symposium for Young Investigators PL 6 Brian T. Murphy University of Illinois at Lake Michigan-Derived Diaza-Anthracene Antibiotics that Target Mycobacterium Chicago, USA Tuberculosis IL 10 Gordon J. Florence University of St Andrews, UK Resolving Structure Through Synthesis: Leiodolide A and Palmerolide C FRIDAY 20th September, 2013

Special Symposium for Young Investigators IL 11 Antonio H. Daranas OC 21 Robert Britton OC 22 Robert Brkljaca OC 23 Efstathia Ioannou OC 24 Kine Østnes Hanssen OC 25 Paula Jimenez

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University of La Laguna, Spain Structural Studies on Complex Polyketides from Dinoflagellates Simon Fraser University, Canada Rmit University, Australia

Chlorohydrins: Versatile Building Blocks for the Synthesis of Marine Natural Products Phytochemical profiling and Biological Activity Studies of the Southern Australian Marine Alga, Sargassum Paradoxum University of Athens, Greece New Polyketides from a Marine-Derived Actinobacterium University of Tromsø, Norway Introducing Atomic Force Microscopy in structure elucidation - The Discovery of Breitfussin A and B, Highly Modified Halogeneted Dipeptides from the Arctic Hydrozoan Thuiaria Breitfussi Labomar - Universidade Cytokinesis blockage induced by a Dithiolpyrrolone isolated from a Federal do Ceara, Brazil Streptomyces sp. Strain recovered from the Brazilian Endemic Tunicate Eudistoma Vannamei



Monday 16th September, 2013

Marine Natural Products: Celebrating Success Marine Natural Products: Celebrating Success MORNING SESSION: Marketed Marine Natural Products: Success stories Chair: Murray Munro 10:05 - 10:50 L 1 Bruce A. Littlefield Eisai, USA 10:50 - 11:20 Coffee break 11:20 - 12:05 L 2 José Mª Fernández Sousa-Faro PharmaMar, Spain 12:05 - 12:50 L 3 Andreas Grassauer Marinomed, Austria 12:50 - 13:35 L 4 Ana Martins Bioalvo, Portugal 13:35 - 15:00 Lunch

Supply Challenge: Marine Natural Products as potential therapeutic agents AFTERNOON SESSION: Marine Natural Products in Development Chair: Ian Paterson 15:00 - 15:40 PL 1 William Fenical 15:40 - 16:10 IL 1 Raymond Andersen 16:10 - 16:40 IL 2 Simon Munt 16:40 - 17:00 Coffee break 17:00 - 17:20 OC 1 Julian Adams 17:20 - 17:40 OC 2 Laurent Meijer 17:40 - 18:00 OC 3 Heonjoong Kang 18:00 - 18:15 Fernando Albericio POSTER SESSION 1 - Hotel Hesperia 18:30 - 21:30 Posters 20:00 Refreshments

University of California, San Diego, USA University of British Columbia, Canada PharmaMar, Spain Infinity Pharmaceuticals, USA Manros Therapeutics, France Seoul National University, South Korea Marine Drugs. e-conference platform Sciforum


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L1

Morning Session

LECTURE 1

DEVELOPMENT OF ERIBULIN, A MACROCYCLIC KETONE ANALOG OF HALICHONDRIN B, FOR ADVANCED BREAST CANCER Bruce A. Littlefield Distinguished Scientist, Oncology, Eisai Inc. 4 Corporate Drive Andover, Massachusetts, USA Email: [email protected]

16th September, 2013

Natural products from marine sources provide a rich source of complex, biologicallyactive compounds as starting points for drug discovery. Related to their roles in chemical defense, marine natural products often have remarkable structural complexity, reflecting eons of chemical evolution for target selectivity. Similarly, their often high biological activity reflects potencies required in highly diluting ocean environments. One such natural product is halichondrin B (HB), isolated from the sponge Halichondria okadai. Reports of HB’s remarkable antitumor activity led to significant interest in developing this agent as a new cancer therapeutic. Although limited supply from sponges prevented such development, the total synthesis of HB and the discovery that its biological activity resided in its smaller “right half” macrolactone moiety provided opportunities to develop a simplified fully synthetic molecule. Eribulin is a synthetic macrocyclic ketone analog of HB’s macrolactone right half. As a tubulintargeted microtubule dynamics inhibitor, eribulin shows low- to sub-nM potency against cancer cells in vitro, disrupting mitotic spindles and leading to apoptosis after 10-12 hours of irreversible mitotic block. In vivo, eribulin induces rapid tumor regression and long-term survival of nude mice bearing human tumor xenografts. Eribulin (as Halaven®) is currently in clinical use in the European Union, United States, Japan and several other countries for treatment of certain advanced breast cancer patients.

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Morning Session

L2

LECTURE 2

DISCOVERY AND DEVELOPMENT OF TRABECTEDIN José María Fernández-Sousa Faro Zeltia, Plaza Descubridor Diego Ordás, 3 - 5º, 28003 Madrid, Spain Email: [email protected]


Ecteinascidia turbinata (family Perophoridae) is a colonial ascidian species found in the Caribbean and the Mediterranean. Aqueous ethanol extracts of Ecteinascidia turbinata were shown to have antitumor effects in 1969, but it was not until 1990 that isolation and structural characterization of the ecteinascidins was completed by Rinehart and coworkers. ET-743 was the most abundant member of the family (0.0001% yield). Early in vitro studies at PharmaMar and the National Cancer Institute (NCI) in a panel of 60 human tumor cell lines confirmed the potent activity of ET-743 (1 pM to 10 nM). Furthermore, the NCI COMPARE analysis using more than 100 standard anticancer agents was negative, indicating a new mechanism of action for ET-743. Later preclinical studies provided further important insights to help design subsequent clinical trials. For early clinical development, Ecteinascidia turbinata was successfully grown and harvested in aquaculture facilities located along the Mediterranean coast and ET-743 isolated by extraction and purification on an industrial scale. In recent years several total syntheses have been published. However, the length and complexity of these processes presented a tremendous barrier to their use for large-scale manufacture of ET-743, especially given the regulatory and quality requirements for pharmaceuticals. The supply problem was finally solved at PharmaMar by development of a semisynthetic procedure suitable for industrial production of the drug. Today, Trabectedin (ET-743), tradename Yondelis® , is sold in more than 80 countries around the world for the treatment of soft tissue sarcoma and ovarian cancer.


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L3

Morning Session

LECTURE 3

ANTIVIRAL MARINE BIOPOLYMERS – FROM CONCEPTS TO THE MARKET Andreas Grassauer Marinomed Biotechnologie Gmbh, Veterinaerplatz 1, 1210, Vienna, Austria. E-mail: [email protected]


Seaweed and polymers derived from seaweed have been used for centuries as food and additive for cosmetics and pharmaceuticals. Efforts to establish sulfated antiviral polymers as blocking agents for sexually transmitted HIV -1infections showed limited success in clinical trials. Recently, Iota -Carrageenan a polymer derived mainly from the species Eucheuma and Chondrus the so-called Irishmoss was recently shown to bear antiviral activity against a broad range of respiratory viruses including rhinoviruses, influenza virus and many others. This resulted in the development of products capable of creating a protective physical barrier in the nasal cavity with an antiviral polymer that works as inhibitor against virus entry for prophylaxis and therapy of respiratory infections. Independent double-blind placebo controlled clinical trials in more than 450 adults and children revealed that the intranasal administration of Iota-Carrageenan with a nasal spray is safe, significantly reduced the time to disease clearance of patients with common cold and decreased the virus load in nasal lavages. This was also true for subgroups of patients infected with influenza A virus, rhinoviruses or coronaviruses. Moreover, treatment with Iota-Carrageenan resulted in a significant reduction of relapses and a significantly higher rate of symptom free patients at the end of the study periods. Intranasal administration of Iota-Carrageenan is currently the only broad band antiviral therapeutic option available on the market for prophylaxis and therapy of respiratory infections.

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Morning Session

L4

LECTURE 4

A NEW BOTULIN-LIKE NEUROMODULATOR BIOACTIVE DERIVED FROM A DEEP SEA MICROORGANISM: APPLICATIONS FOR DAILY SKINCARE Martins, A. 1, Rodrigues, I. 1, Almeida, A. 1, Rodrigues, C. 1, Pinheiro, R. 1, Calado, P. 1 , Vieira, H. 1,2. 1


BIOALVO S.A., TEC LABS - CENTRO DE INOVAÇÃO - Campus da FCUL, Campo Grande, 1748016 Lisboa, Portugal. 2University of Lisbon, Faculty of Sciences, Centre for Biodiversity, Functional and Integrative Genomics(BioFIG),-Campus da FCUL, Campo Grande, 1748-016 Lisboa, Portugal. E-mail:[email protected]

Screening of phylogenetically diverse and unique microorganisms from rare or extreme ecosystems has increased tremendously in the last decade with relevant bioactivities being discovered. In this work, a marine prokaryotic collection, called PharmaBUG, was isolated from four deep sea hydrothermal vents in the Mid-Atlantic Ridge near Azores islands, Portugal. This collection was highly characterized and tested for daily skincare applications. RefirMARTM ingredient was identified as a novel marine microorganism derived bioactive that inhibits the release of acetylcholine at the synapse of the motor neuron. Interestingly, and in contrast to the well known botulinum toxin (BOTOX®), RefirMARTM does not have a permanent action in the motor neuron but instead inhibits the release of acetylcholine vesicles in a transient way (in vitro dosage range of 5 mg/ml to 10 mg/ml) making it highly advantageous. This natural neuromodulator is a crude protein extract highly soluble in water, safe, stable and noncytotoxic. In addition, RefirMARTM shows dermal penetration capacity and thus, potential for topical application, avoiding the inconvenience of administration by injection and medical supervision which offers limitless applications for cosmetics. A 28-day clinical study in 12 volunteers was performed to assess RefirMAR’sTM efficacy in wrinkle smoothing and line reduction, upon daily usage of a 1% formulation. Interestingly this trial has shown an up to 23% wrinkle depth decrease and an up to 64% hydration increase. RefirMARTM is thus the first success story in BIOALVO’s path towards marine microorganism-derived added value products.


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PL 1

Afternoon Session

PLENARY LECTURE 1

SERINIQUINONE, A MARINE MICROBIAL METABOLITE FOR THE TREATMENT OF MELANOMA William Fenical and Co-workers Center for Marine Biotechnology and Biomedicine. Scripps Institution of Oceanography University of California, San Diego. La Jolla, CA 92093-0204 E-mail: [email protected]


Melanoma is one of the fastest growing of all cancers with an estimated 48,000 deaths reported each year. While less common than other skin cancers, metastatic melanoma is much more dangerous and difficult to treat, thus developing new therapies is of great significance. We examined a unicellular Gm + actinobacterium, identified as a member of the genus Serinicoccus, that was recovered from shallow marine sediments. In culture, this strain produced seriniquinone, a bright orange metabolite with very limited solubility. In vitro screening indicated modest cytotoxicity toward HCT-116 colon carcinoma, however in the NCI 60 cell line panel seriniquinone showed pronounced selectivity toward all of the melanoma cell lines tested. Subsequent biochemical and pharmacological studies have shown that seriniquinone targets a poorly known protein that is highly overexpressed in melanoma. Targeting this protein activates apoptosis and the induction of autophagocytosis, leading to efficient cell death.

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Afternoon Session

IL 1

INVITED LECTURE 1

DISCOVERY AND SYNTHESIS OF MARINE NATURAL PRODUCT DRUG LEADS Raymond Andersen Department of Chemistry, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z1 E-mail: [email protected]



The secondary metabolites found in marine organisms represent an extremely rich source of novel chemical diversity for academic drug discovery and chemical biology programs. Marine sponges and marine microorganisms continue to be the most prolific sources of new bioactive marine natural products. Our group at UBC has amassed a sizable library of crude extracts from marine sponges and cultures of marine microorganisms collected in many of the world's oceans. In collaboration with biologists, this crude extract library has been screened for activity in a variety of cell-based and pure enzyme assays designed to identify promising lead compounds for the development of drugs. Bioassay-guided fractionation of the crude extracts and extensive spectroscopic analysis is used to identify the structures of the pure natural products active in these assays. Biology oriented chemical synthesis is used by our group to probe the SAR for new natural product pharmacophores that we discover and to provide material for in vivo testing in animal models. Several new drug candidates for the treatment of cancer, inflammation, cystic fibrosis, and infectious diseases have emerged from this research program. Three of them have progressed to phase II clinical trials in humans and others are in preclinical evaluation/development. The lecture will present recent chemical and biological results from our academic drug discovery/chemical biology research

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IL 2

Afternoon Session

INVITED LECTURE 2

ISOLATION, SYNTHESIS, PRECLINICAL AND CLINICAL DEVELOPMENT OF PM060184 Simon Munt, María Jesús Martín, Laura Coello, Rogelio Fernández, Fernando Reyes, Alberto Rodríguez, Carmen Murcia, María Garranzo, Cristina Mateo, Francisco Sánchez-Sancho, Santiago Bueno, Carlos de Eguilior, Andrés Francesch and Carmen Cuevas R&D, PharmaMar, S. A., 28770 - Colmenar Viejo, Madrid, Spain E-mail: [email protected]


Microtubules continue to be one of the most successful anticancer drug targets and a favourite hit for many naturally occurring molecules. Whilst two of the most successful representative agents in clinical use, the taxanes and the vinca alkaloids, come from terrestrial sources, the sea has also proven to be a rich source of new tubulin-binding molecules. We describe herein the first isolation, structural elucidation and total synthesis of two totally new polyketides isolated from the Madagascan sponge Lithoplocamia lithistoides. Both PM050489 and PM060184 show antimitotic properties in human tumor cells lines at subnanomolar concentrations and display a distinct inhibition mechanism on microtubules. The development of an efficient synthetic procedure has solved the supply problem and, following pharmaceutical and preclinical development, has allowed PM060184 to start clinical studies as a promising new drug for cancer treatment.

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Afternoon Session

OC 1

ORAL COMMUNICATION 1

HSP90 INHIBITORS IN THE CLINIC: RETASPIMYCIN HCL (IPI-504) IN NON-SMALL CELL LUNG CANCER (NSCLC) Julian Adams Infinity Pharmaceuticals, 780 Memorial Drive, 02139, Cambridge, Estados Unidos. E-mail: E-mail: [email protected]



Heat-shock protein 90 (Hsp90) is an attractive target in cancer therapy due to its role in maintaining the activity and stability of a variety of oncoproteins. Hsp90 inhibitors disrupt multiple oncogenic signals. Retaspimycin HCl (IPI504), a potent and selective Hsp90 inhibitor, is being studied in NSCLC. Retaspimycin HCl has shown preclinical activity both as a single agent and in combination in mouse models of cancer. The molecule has an ansamycin backbone, based on with the natural product, geldanamycin. The natural product itself was sub-optimal with respect to safety and drug delivery. Retaspimycin HCl is the hydroquinone derivative of 17-allylamino geldanamycin which is both air and pH sensitive. Following initial Phase 1/2 development as a single agent, retaspimycin HCl is currently being explored in two combination trials in non-small cell lung cancer (NSCLC). The first trial is a, randomized, placebo-controlled Phase 2 trial, testing the combination of retaspimycin HCl with docetaxel in patients with NSCLC. The rationale for this trial is a Phase 1b trial of that showed the combination was well tolerated and resulted in an overall response rate of 26% (n=23) which compares favorably to historical data with docetaxel as a single agent (ORR ~8%) in that population (Riely, ASCO 2011).The second investigation is a Phase 1 trial evaluating the combination of retaspimycin HCl with everolimus in RAS mutated NSCLC; this trial is based on encouraging preclinical results of the combination of retaspimycin HCl/rapamycin in a mutant KRAS/p53 mouse lung cancer model (De Raedt, Cancer Cell 2011).

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OC 2

Afternoon Session


LEUCETTINES, A FAMILY KINASE INHIBITORS DERIVED FROM THE MARINE SPONGE ALKALOID LEUCETTAMINE B. DEVELOPMENT AS DRUG CANDIDATE FOR DOWN SYNDROME AND ALZHEIMER"S DISEASE 1

2

3

3

Laurent Meijer , Tania Tahtouh , Francois Carreaux , Jean-Pierre Bazureau , Arnaud 5 1 4 4 5 Duchon , Yann Herault , Jonathan Elkins , Stefan Knapp , Emilie Durieu 1


Manros Therapeutics, Manros Therapeutics, Centre De Perharidy, 29680, Roscoff, Francia 2CNRS, Roscoff, 29680, France. 3UNIVERSITY OF RENNES, Rennes, 35000 , France 4Institut De Génétique Et De Biologie Moléculaire Et Cellulaire, Illkirch, 67404, France.5University Of Oxford, Structural Genomics Consortium, Oxford, Ox3 7dq, Uk E-mail: [email protected]

There is growing evidence implicating DYRKs (dual specificity, tyrosine phosphorylation regulated kinases) and CLKs (cdc2 -like kinases) in the onset and development of neurodegenerative pathologies such as Alzheimer’s disease and Down syndrome. A random screen of natural products has allowed us to identify the marine sponge Leucettamine B as an inhibitor of DYRKs and CLKs. Synthesis of analogues (collectively referred to as Leucettines) led to an optimized product, Leucettine L41. L41 was co-crystallized with DYRK1A, DYRK2, CLK3 and PIM1. The selectivity of L41 was extensively studied (i) by its effects on the catalytic activity of panels of recombinant kinases, (ii) in an interaction assay with 402 kinases, (iii) by an affinity chromatography approach (L41 immobilized on sepharose beads) and (iv) by a competition affinity assay. These approaches revealed unexpected potential secondary targets such as casein kinase 2, STE20-like Ser/Thr Kinase and phosphatidylinositol-3-phosphate 5-kinase type III and associated Vac14/Fig4. Proteomic and transcriptomic studies shed light on the range of molecular actions of L41 in cells. Leucettine L41 displayed dose-and time-dependent neuroprotective effects on glutamate-induced cell death in immortalized mouse hippocampal cell line HT22. L41 also reduced amyloid precursor protein -induced cell death in cultured rat brain slices. L41 is currently being tested in the tgBACDyrk1a mice model which expresses one extra copy of the DYRK1A gene and shows cognitive deficits. The unusual multi-target selectivity of Leucettines may account for their neuroprotective effects. This family of kinase inhibitors deserves further optimization as potential therapeutics against Alzheimer’s disease and Down syndrome. References 1.

2.

20

Tahtouh, T., Elkins, J.M., Filippakopoulos, P., Soundararajan, M., Burgy, G., Durieu, E., Cochet, C., Schmid, R., Lo, D.C., Delhommel, F., Oberholzer, A.E., Pearl, L.H., Carreaux, F., Bazureau, J.P., Knapp, S. and Meijer, L., 2012. Selectivity, co-crystal structures and neuroprotective properties of Leucettines, a family of protein kinase inhibitors derived from the marine sponge alkaloid Leucettamine B. J. Med. Chem. 55, 9312-9330. Debdab, M., Carreaux, F., Renault, S., Soundararajan, M., Fedorov, O., Filippakopoulos, P., Lozach, O., Babault, L., Tahtouh, T., Baratte, B., Ogawa, Y., Hagiwara, M., Eisenreich, A., Rauch, U., Knapp, S., Meijer, L. and Bazureau, J.-P., 2011. Leucettines, a class of potent inhibitors of cdc2-like kinases and dual specificity, tyrosine phosphorylation regulated kinases derived from the marine sponge leucettamine B. Modulation of alternative pre-RNA splicing. J. Med. Chem. 54, 4172-4186.



Afternoon Session

OC 3


POTENTIAL DRUG LEADS FOR DIABETES, OSTEOPOROSIS AND BACTERIAL INFECTION FROM MARINE ORGANISMS Hiyoung Kim,1 Shin-Young Ryu,2 Sunhee Park,1 Giri Awadut,1 Yallamalla Srinivas,1 Rahul Shivaji Patil,1 Weihong Wang,1 Youn-Jin Son, 3 Won-Kyung Ho,2 and Heonjoong Kang1,* 1

Center for Marine Natural Products and Drug Discovery, School of Earth and Environmental


Sciences, College of Natural Sciences; 2Department of Physiology, Biomembrane Plasticity Research Center and Neuroscience Research Institute, College of Medicine, Seoul National University, Seoul, 110-799; 3Suncheon Research Center for Natural Medicines, College of Pharmacy, Sunchon National University, Suncheon, Jeonnam 540-950, Korea E-mail: [email protected]

Obesity and its associated diabetes are becoming more and more of a threat to public health. Type II diabetes mellitus (T2DM) is characterized by high blood glucose as a consequence of insulin resistance and relative insulin deficiency. Type II diabetes is treated by several approaches: insulin secretagogues (e.g., sulfonylureas), insulinsensitizing agents (e.g., metformin and thiazolidinediones) and -glucosidase inhibitors. Particularly, sulfonylureas, the KATP channel inhibitors of the pancreatic -cells which help the release of insulin, until now were widely used as oral anti-diabetics. The KATP channel plays a major role in controlling the -cell membrane potential. But, the KATP channel blockers cause a high risk of hypoglycemic events. Recently, it was discovered that the modulation of voltage-gated potassium channels may be an alternative in yielding anti-diabetic indications. The inhibition of -cell Kv currents prolongs the action potentials, sustains the opening of voltage-dependent Ca2+ channels, and thereby enhances glucose-induced insulin release without causing risky hypoglycemia. Thus, the -cell Kv channel has attracted much attention as a potential therapeutic target for the treatment of type II diabetes. Through screening of bioactive marine natural products against the Kv channel, we found a couple of compounds to inhibit the Kv channel with IC50 of 0.59 μM. Additionally, we also discovered novel hits for treatment of osteoporosis and novel antibiotic leads against MRSA, VRE and VISA. Their structural elucidation, total synthesis and structural modification including bioactivities will be discussed.


21



Tuesday 17th September, 2013

Supply Challenge: Marine Natural Products as potential therapeutic agents Supply Challenge: Marine Natural Products as potential therapeutic agents MORNING SESSION: Chemical Synthesis Chair: Tadeusz Molinski 09:00 - 09:40 PL 2 Ian Paterson 09:40 - 10:10 IL 3 Jef K. De Brabander 10:10 - 10:30 OC 4 Brent Copp 10:30 - 10:50 OC 5 Thomas Lindel 10:50 - 11:30 Coffee break 11:30 - 11:50 OC 6 Nicolas Inguimbert 11:50 - 12:10 OC 7 Marta Pelay Gimeno 12:10 - 12:30 OC 8 Daniel Romo 12:30 - 13:00 IL 4 Angelo Fontana 13:00 - 14:30 Lunch AFTERNOON SESSION: Biosynthesis: the "omics" era Chair: Vassilios Roussis 14:30 - 15:10 PL 3 Bradley Moore 15:10 - 15:40 IL 5 Jörn Piel 15:40 - 16:00 OC 9 Letícia V. Costa-Lotufo 16:00 - 16:20 OC 10 Brandon I. Morinaka 16:20 - 17:00 Coffee break 17:00 - 17:20 OC 11 Toshiyuki Wakimoto 17:20 - 17:40 OC 12 Nadine Ziemert 17:40 - 18:10 IL 6 Pei-Yuan Qian POSTER SESSION 2 - Hotel Hesperia 18:30 - 21:30 Posters 20:00 Refreshments

University of Cambridge, UK UT Southwestern Medical Center, USA University of Auckland, New Zealand TU Braunschweig, Germany University of Perpignan, France Institute for Research in Biomedicine, Spain Texas A&M University, USA CNR - Istituto di Chimica Biomolecolare, Italy

Scripps Institution of Oceanography, USA Eidgenössische Technische Hochschule Zurich, Switzerland Universidade Federal do Ceara, Brasil Eidgenössische Technische Hochschule Zurich, Switzerland The University of Tokyo, Japan Scripps Institution of Oceanography, USA Hong Kong University of Science & Technology, China


23


PL 2

Morning Session

PLENARY PL ENARY LECTURE LECTURE 2

CHALLENGES C HALL LENGES A AND ND D DISCOVERIES ISCOVERIES IN IN MARINE MARINE MACROLIDE MACROLIDE SYNTHESIS SYNTHES SI S IIan aan Paterson Patterson Department De partment of of Chemistry, Chemistry, University U iversity of Un of C Cambridge, a m b r id g e , C Cambridge a m b rid g e C CB2 B2 1EW, 1EW, UK E Email: ip [email protected] E-mail: [email protected]

M arrine na attural products products continue continue to to provide provide an an unparalleled unparralleled opportunity opportunity for for o driving d ving dri Marine natural cr eattivity aand nnd di scovery. Recent Recent w orrk from from r will bee presented presented directed directed towards towards r the the creativity discovery. work our group will ttotal otal ssynthesis ynthe h sis of rare raarre and annd architecturally architecturally complex com mpplex marine marrine macrolides macrolides having haavving promising prom mising [1]] [1 aanticancer nnticanncer activity, activity, iincluding ncluding lleiodermatolide eiodermatolide (i (isolated solatted ffrom rom r tthe he llithistid ithistid sponge sponge p


[2]] Leiodermatium L eiodermatium ssp.) p.)[2 and annd the the aplyronines applyroni y nes (isolated (isolated from from r the the Japanese Jappannese sea sea hare harre Aplysia Apl plysia [3]] [3 kkurodai). urodai). Each Each of the the h targets targets selected selected presents presents a unique unique set set of challenges, challenges, inspiring inspiring n the the

de development velopment of new new methods methods and and n strategies strattegies to to circumvent circum mvent the the multiple multiple problems problems eencountered. ncountereed. On On a wider wider level, level, our work work has has led led to to the the development developm o ent of versatile versatile and annd general geeneral aapproaches ppproaches for for o the the stereocontrolled stereocontrolled construction construction of the the characteristic chaarracteristic molecular molecular scaffolds scaffol o ds aassociated ssociated with with a large larrge variety vaarriety of important importannt polyketides polyketides off marine marrine and annd terrestrial terrestrial origin, o gin, ori biological eenabling naabl b ing ttheir h ir ssustainable he ustainable ssupply upply ffor or o ffurther urt u her bi ological sstudies. tudies.

References R eferences 1 a) a Dalby, Dalby, S. S. M.; M.; Paterson, Paterson, I. I. Curr. Cuurr. O Opin. p in . D Drug rug Discovery Discovery Dev. Dev. 2010, 13, 13, 7777; 77; b) b) Yeung, Yeung, K.-S.; K.-S S.; Paterson, P Pa terson, I. I. Chem. Chem. Rev. Rev. 2005, 105, 105, 44237; 237; c) c) Paterson, Paterson, I.; I.; Anderson, Andeerson, E. E. A. A. Science Science 2005, 310, 4451. 51. 2 Paterson, P terson, I.; Pa I.; Dalby, Dalby, S. M.; M.; Roberts, Roberts, J. J. C.; C.; Naylor, Naylor, G. G. J.; J.; Guzman, Guzman, E. E. A.; A.; Isbrucker, Isbrucker, R.; R.; Pitts, Pitts, T. T. P.; P.; Linley, Ed. Linley, P.; P.; Divlianski, Divlianski, D.; D.; Reed. Reed. J. J. K.; K.; Wright, Wright, A. A. E. Angew. Angew. Chem. Chem. Int. Int. Ed d. 2011, 50, 50, 33219. 219. 3 a) Yamada, Am. Yamada, K.; K.; Ojika, Ojika, M.; M.; Ishigaki, Ishigaki, T.; T.; Yoshida, Yoshida, Y.; Y.; Ekimoto, Ekimoto, H.; H.; Arakawa, Arakawa, M. M. J. A m. Chem. Chem m. Soc. Soc. 1993, 19933, 115, 115, 111020; 1020; b) b) Ojika, Ojika, M.; M.; Kigoshi, Kig i oshi, H.; H.; Suenaga, Suenaga, K.; K.; Imamura, Imamura, Y.; Y.; Yoshikawa, Yoshikawa, K.; K.; Ishigaki, Ishigaki, T.; T.; Sakakura, Sakakura, A.; A.; Mutou, Mutou, T.; T.; Yamada, Yamada, K. K. Tetrahedron Tetrahedron 2012, 68, 68, 982. 982.

24



Morning Session

IL 3

INVITED LECTURE 3

NATURAL PRODUCTS: AN OPPORTUNITY FOR DISCOVERY RESEARCH Jef De Brabander Ut Southwestern Medical Center, Department Of Biochemistry, 5323 Harry Hines Blvd, Texas, Tx 75390-9038, Dallas, Estados Unidos. E-mail: [email protected]



Research in De Brabander laboratory focuses on the synthesis of complex molecular architectures. Synthetic targets include both designed and naturally occurring substances with novel structural features and interesting biological function. To facilitate the development of efficient and practical syntheses, the De Brabander group develops methodology for the construction of functionality found in complex natural products and small molecules of interest to drug discovery programs. Taking advantage of the research environment at UT Southwestern Medical Center, his group has integrated molecular pharmacology and biochemical studies to understand and create biologically active small molecules.

25


OC 4

Morning Session


THE DISCOVERY AND DEVELOPMENT OF NEW ANTIMALARIAL DRUG LEADS FROM LIBRARIES OF ASCIDIAN NATURAL PRODUCTS Brent Copp University of Auckland, School of Chemical Sciences, Private Bag 92019, 1142, Auckland, New Zealand. E-mail:[email protected]


Parasitic protozoal infection is a cause of suffering for an estimated one billion people worldwide. Insect bites that transfer protozoa to human hosts lead to the diseases malaria (protozoa: Plasmodium falciparum), Chagas Disease (also known as American Trypanosomiasis, Trypanosoma cruzi), Leishmaniasis (Leishmania spp.) and Sleeping Sickness (also known as Human African Trypanosomiasis, Trypanosoma brucei rhodesiense, T. brucei gambiense). While drugs exist for these diseases, they have undesirable side-effects or are showing reduced efficacy due to the growing prevalence of drug resistant strains. In the context of our continuing search for new leads for the development of treatments for neglected human diseases, we recently initiated a program of screening a library of synthesized and isolated marine natural products against a panel of four parasitic protozoa: Trypanosoma brucei rhodesiense, Trypanosoma cruzi, Leishmania donovani and Plasmodium falciparum K1 dual drug-resistant strain, with concurrent counterscreening for toxicity towards the non-malignant L6 rat myoblast cell line. A diverse range of hits were identified, with many of the natural products originating from ascidians. For example orthidine F (1), didemnidine B (2), ascidiathiazone A (3), eudistomin X (4), scabellone B (5) and opacalines B and C (6 and 7) were all found to be relatively non-toxic, moderate growth inhibitors of a dual drug-resistant strain of Plasmodium falciparum with IC50 0.89 μM, 15.0, 3.3, 1.9, 4.8, 2.5 and 4.5 μM, respectively. Extensive structure-activity relationship studies of the four compound classes represented by 1-4 have yielded analogues with enhanced in vitro potency (IC50 low to sub-nanomolar) and excellent selectivity. Follow-up in vivo studies against P. berghei using either ip or oral dosing has identified a number of active examples, highlighting their potential to act as antimalarial drug leads. The synthesis of the target compounds and discussion of biological data in a SAR context will be presented.

26



Morning Session

OC 5


NOVEL ACCESS TO CUBITANE-AND EUNICELLANE-TYPE DITERPENOIDS Thomas Lindel, Kristina Simon, Johannes Wefer, Mona Al Batal T U Braunschweig, Institute of Organic Chemistry, Hagenring 30, 38106, Braunschweig, Germany E.mail: [email protected]

Cubitane-and bicyclic eunicellane-type diterpenoids constitute very interesting natural products from marine corals. Examples include klysimplexin R (1) from Klyxum simplex[1] and calyculone A (2) from Eunicea calyculata.[2] Recently, cytotoxicity of calyculone A (2) against several human cancer cell lines in the higher nanomolar range was reported.[3] The first cubitanoid was cubitene (3), isolated from the secretion gland of soldiers of the East African termite Cubitermes umbratus.[4]


The biosynthesis of both skeletons probably proceeds via cembrane-type precursors, which undergo ring contraction or cyclization, respectively. In this communication it is shown that the chemical synthesis of the title skeletons can also start from menthanetype building blocks, which are accessible in a much more facile manner than cembranoids. Key reagent in both cases is samarium diiodide which is employed in Barbier-type bicyclizations[5] and pinacol couplings.[6]

References 1. 2. 3. 4. 5. 6. 7.

Chen, B.-W.; Chao, C.-H.; Su, J.-H.; Tsai, C.-W.; Wang, W.-H.; Wen, Z.-Z.; Huang, C.-Y.; Sung, P.-J.; Wu, Y.-C.; Sheu, J.-H. Org. Biomol. Chem. 2011, 9, 834-844. Look, S. A.; Fenical, W.; Zheng, Q. T.; Clardy, J. J. Org. Chem. 1984, 49, 1417-1423. Wei, X.; Nieves, K.; Rodríguez, A. D. Pure Appl. Chem. 2012, 84, 1847-1855. Prestwich, G. D.; Wiemer, D. F.; Meinwald, J.; Clardy, J. J. Am. Chem. Soc. 1978, 100, 25602561. Simon, K.; Wefer, J.; Schöttner, E.; Lindel, T. Angew. Chem. Int. Ed. 2012, 51, 10889-10892. Al Batal, M.; Jones, P. G.; Lindel, T. Eur. J. Org. Chem. 2013, online.


27


OC 6

Morning Session


REVISION OF THE LAXAPHYCIN B STRUCTURE BASED ON ITS SYNTHESIS AND POSSIBLE GENERALISATION TO ITS ANALOGS Nicolas Inguimbert, France Boyaud, Isabelle Bonnard, Bernard Banaigs LCBE EA 4215, 58 Avenue P Alduy, Centre de Phytopharmacie, Lcbe Ea 4215, 66860, Perpignan, France. E-mail: [email protected]


Considering that the cyclic and cytotoxic dodecapeptide laxaphycin B[1]-[2] and its analogs lobocyclamides[3] and lyngbyacyclamides[4] are all produced by cyanobacterias we hypothesized that they are produced by a similar biosynthetic pathway. Consequently, they are highly homologous if we consider their amino acids sequences. We could also expect that this will be the case for the stereochemistry of the amino acids entering in their sequences but in fact subtle variations at the stereochemical level are reported. Therefore, one way to confirm their structures relies on the development of a synthesis offering some flexibility in terms of amino acids mutation. We will report on the first total synthesis of laxaphycin B that was accomplished through stepwise automated Solid Phase Peptide Synthesis (SPPS), leading to the structure revision of laxaphycin B especially in regards with the configuration of one of the two 3-hydroxyleucines that entered in its composition. The analogous lyngbyacyclamide A was obtained by an extension of this synthesis. This synthesis offered the possibility to check through a structure activity relationships study a way to identify the key amino acids responsive for their cytotoxicity.

References 1. 2. 3. 4.

28

Frankmölle, W. P.; Knübel, G.; Moore, R. E.; Patterson, G. M. J. Antibiot. (Tokyo) 1992, 45, 1458-1466. Bonnard, I.; Rolland, M.; Francisco, C.; Banaigs, B. Lett. Pept. Sci. 1997, 4, 289-292. MacMillan, J. B.; Ernst-Russell, M. A.; de Ropp, J. S.; Molinski, T. F. J. Org. Chem. 2002, 67, 8210-8215. Maru, N.; Ohno, O.; Uemura, D. Tetrahedron Lett. 2010, 51, 6384-6387.



Morning Session

OC 7


FIRST TOTAL SYNTHESIS OF THE ANTITUMORAL MARINE CYCLODEPSIPEPTIDE PIPECOLIDEPSIN A 1

2

1

2

Marta Pelay Gimeno , María Jesús Martin , Jan Spengler , Andrés Francesch , 2 1 1 Carmen Cuevas , Judit Tulla-Puche , Fernando Albericio 1

Institute for Research in Biomedicine, IRB Barcelona, Baldiri Reixac, 10, 08028 - Barcelona, Spain. 2 PharmaMar, S. A., Avda. de los Reyes, 1, 28770 - Colmenar Viejo, Madrid, Spain. E-mail: [email protected]



During the last decades the literature has been enriched with a new class of natural products named “head-to-side-chain” cyclodepsipeptides. All of them are produced by marine sponges and show relevant biological profiles, mostly including cytotoxic and anti HIV activities. A number of structural features are shared by all these peptides. Firstly, the defining “head-to-side-chain” scaffold via an ester bond between the Cterminal and a beta-hydroxy group. Secondly, a branching position consisting of a Dallo-beta-hydroxy alpha-amino acid, which is always acylated by the same rare residue. Finally, the presence of certain unique amino acids and an N-terminal blocked with a beta-hydroxy acid. Callipeltins, neamphamides, papuamides, theopapuamides, mirabamides and homophymines have already been described in the literature. Herein, we report for the first time the synthesis of the potent antineoplastic agent pipecolidepsin A, a “head-toside-chain” cyclodepsipeptide bearing the unprecedented and extraordinary bulky AHDMHA residue at the branching point. The construction of the extremely hindered ester bond and the synthesis and assembly of the complex moieties (3S,4R)-3,4dimethyl-L-glutamine; (2R,3R,4S)-4,7-diamino-2,3-dihydroxy-7-oxoheptanoic acid, DADHOHA; L-threo-beta-EtO-Asn; and (2R,3R,4R)-3-hydroxy-2,4,6trimethylheptanoic acid, HTMHA represent the major synthetic challenges overcome.

29


OC 8

Morning Session


INNOVATIONS IN SYNTHETIC EFFICIENCY AND NATURAL PRODUCT DERIVATIZACION DRIVEN BY NATURAL PRODUCTS AND IMPACTING CELL BIOLOGY Daniel Romo Department of Chemistry and the Natural Products LINCHPIN Laboratory, Texas A&M University, College Station, TX 77842-3012, USA. E-mail: [email protected]


Synthetic chemists are challenged in the 21st century by the goal of developing more efficient, scaleable synthetic designs to access complex molecules including natural products with an eye towards mining their rich information content for basic cell biology. Our group has been engaged in these endeavors and this talk will describe our recent discoveries surrounding the potent antitumor agent, agelastatin A, a member of the pyrrole-2-aminoimidazole marine alkaloid family. In addition, the application of novel derivitization strategies and organocatalytic, rapid complexity-building synthetic methods applicable to bioactive natural products will be described.

30



Morning Session

IL 4

INVITED LECTURE 4

BIOACTIVE METABOLITES FROM MARINE PROTISTS A. Fontana1, A. Cutignano1, R. De Palma2, G. d’Ippolito1, E. Manzo1, G. Nuzzo1, D. Pagano1 1

Istituto di Chimica Biomolecolare, Consiglio Nazionale delle Ricerche Via Campi Flegrei 34, 80078 Pozzuoli, Napoli, Italy. 2Department of Clinical and Experimental Medicine, Second University of Naples, Via S. Pansini 5 (Edif. 3), 80131 Napoli, Italy E-mail:[email protected]


Although decades of pioneering studies in marine microbiology have demonstrated that marine microorganisms have great potential as source of novel bioactive molecules, conspicuous invertebrates from temperate and tropical waters remain the main origin of compounds described in the specialized literature.[1] However, the natural concentrations of many pharmacologically active compounds in these organisms are often minute and sometimes account for less than few percentages of the wet weight. In addition, the source species may be comparatively rare, and an adequate supply may require more collections that are often hampered by strict national and international laws that tend to protect the source organism and its habitat from overexploitation. These aspects strongly influence the identification of novel marine bioactive compounds, impacting both the biological and chemical studies especially for marine organisms of remote regions. Here we report our recent studies on bioactive compounds isolated from infrequently encountered marine organisms and unexplored phyla. The contribution is mainly focused on the exploration of marine protists as inspiration and source of novel compounds for human wellbeing. We will discuss the technological tools for addressing this task, including the development of novel culture conditions, as well as the chemical and biochemical protocols for their analysis and control. The main body of the report will concern identification of the bioactive natural products, their biotechnological production and preparation of synthetic analogs. Most of the data are related to the results obtained within the frame of two large projects concerning discovery of novel antitumor leads and immunomodulatory agents. References 1. J.W. Blunt, B.R. Copp, R.A. Keyzers, M.H.G. Munro, M.R. Prinsep. Marine natural products. Nat. Prod. Rep. 30, 2013, 237-323.


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PL 3

Afternoon Session

PLENARY LECTURE 3

MARINE BACTERIAL BIOSYNTHESIS OF POLYBROMINATED ORGANIC COMPOUNDS Bradley S. Moore Scripps Institution of Oceanography E-mail: [email protected]


The identification and tracking of halogenated organic compounds (HOCs) in the marine environment has revealed their widespread distribution in organisms occupying higher trophic levels of the marine food web. Based on chemical analysis of environmental and biological samples, it is known that several classes of toxic HOC compounds persist in the marine environment of both natural and anthropogenic origin. Despite our ability to detect these compounds in the environment, their origin and their entry point into the marine food web is unknown. This seminar will highlight the bacterial origin and biosynthesis of diverse organohalogen compounds that are structurally related to polybrominated diphenyl ethers, polybrominated dibenzodioxins and polybrominated bipyrroles. A biosynthetic model based on in vivo and in vitro studies will be introduced connecting polybrominated phenol and pyrrole-based chemistry in a model marine bacterium to allow for the genetic source tracking of these biosynthetic signatures across trophic levels of the marine food web.

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Afternoon Session

IL 5

INVITED LECTURE 5

SINGLE-CELL-GUIDED IDENTIFICATION OF NATURAL PRODUCT SOURCES IN COMPLEX SPONGE MICROBIOMES Jörn Piel Institute of Microbiology, Eidgenoessische Technische Hochschule (ETH) Zurich, Switzerland. E-mail: [email protected]


Sponges are a rich source of bioactive natural products with significant drug development potential. Since many sponge species harbor highly complex and specific bacterial communities, a possible microbial origin of at least some sponge compounds has been under discussion. For several sponge-derived compounds,1-4 we have provided metagenomic evidence for a bacterial origin, but the taxonomic identity of the producers remained unknown. This talk discusses two single-cell methods to pinpoint natural product sources in sponges: FACS and FluidFM, the latter being a recently developed method based on atomic force microscopy. In a collaborative study with groups at Waseda University and the Universities of Tokyo and Bielefeld, we investigated Theonella swinhoei Y, a chemically extremely prolific sponge that is the source of onnamides, theopederins, polytheonamides, orbiculamides, cycloand pseudotheonamides, nazumamides, and aurantosides. Surprisingly, a single bacterium within the complex microbiome was identified as the producer of all compounds with the possible exception of the aurantosides.5 Genomic analysis showed that the producer coexists with a closely related second variant in this sponge chemotype. Both bacteria possess large genomes with numerous natural product biosynthetic gene clusters, many of which with as-yet unknown function. The search for relatives in other sponges provided evidence for a more widespread role in the chemistry of these animals. Their rich and highly variable natural product repertoire, which for the examined representatives seems to be non-overlapping with that of cultivated bacteria, exhibits significant discovery potential. References 1.

J. Piel, D. Hui, G. Wen, D. Butzke, M. Platzer, N. Fusetani, S. Matsunaga, Proc. Natl. Acad. Sci. U.S.A. (2004) 101:16222. 2. T. Nguyen, K. Ishida, H. Jenke-Kodama, E. Dittmann, C. Gurgui, T. Hochmuth, S. Taudien, M. Platzer, C. Hertweck, J. Piel, Nat. Biotechnol. (2008) 26:225. 3. K.M. Fisch, C. Gurgui, N. Heycke, S.A. van der Sar, S.A. Anderson, V.L. Vebb, S. Taudien, M. Platzer, B.K. Rubio, S.J. Robinson, P. Crews, J. Piel, Nat. Chem. Biol. (2009) 5:494. 4. M.F. Freeman, C. Gurgui, M.J. Helf, B.I. Morinaka, A.R. Uria, N.J. Oldham, H.-G. Sahl, S. Matsunaga, J. Piel, Science, (2012) 338:387. 5. M.C. Wilson, T. Mori, C. Rückert, A.R. Uria, K. Takada, C. Gernert, U. Steffens, N. Heycke, S. Schmitt, C. Rinke, C. Gurgui, M.J. Helf, E.J.N. Helfrich, T. Wakimoto, M. Kracht, A. Semeniuk, M. Crüsemann, U. Hentschel, I. Abe, S. Matsunaga, J. Kalinowski, H. Takeyama, J. Piel, submitted.


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OC 9

Afternoon Session


METABOLOMICS ANALYSIS OF ZOANTHIDS COLLECTED ALONG THE BRAZILIAN COAST SUGGESTS A CORRELATION BETWEEN MICROBIAL CONTENT AND CYTOTOXICITY Letícia V. Costa-Lotufo1,2, Bianca D. B. Sahm1,2, Arinice M. Costa1, Diego V. Wilke1, Paula C. Jimenez2, Francisco C. L. Pinto3, Otília D. L. Pessoa3, Tito M. C. Lotufo2, Norberto P. Lopes4 1

Departamento de fisiologia e farmacología. Universidade Federal do Ceará (UFC), Fortaleza, CE, Brazil. 2 Instituto de Ciências do Mar (UFC), Fortaleza, CE, Brazil. 3Departamento de Química Orgânica e Inorgânica (UFC), Fortaleza, CE, Brazil. 4 Faculdade de Ciências Farmacêuticas, Universidade de Ssão Paulo, Ribeirão Preto, SP, Brazil. E-mail: [email protected]


Brazil has the second largest coastline of the world containing a remarkable diversity in its ecosystems. As part of an anticancer drug discovery and development program in Brazil, we are interested in further exploring the Zoanthidea family, as a potential source for new structural classes. Herein, we extended a metabolomics approach to explore the differences between the cytotoxic activity of the extracts from Protopalythoa variabilis and Palythoa caribaeorum collected in four different locations along Brazilian coast (Paracuru and Taiba beaches, Ceara State; Arraial do Cabo and Búzios; Rio de Janeiro State; and Arvoredo Island, Santa Catarina State). The extracts were analyzed by HR-LC/MS and their cytotoxicity evaluated against two cell lines (PC-3M and HCT-116) using MTT assay. Interestingly, the activity of the extracts did not correlate with species, with both species collected at Paracuru Beach displaying the highest activity (P. variabilis, IC50 = 2.8 μg/mL; P. caribaeorum, IC50 < 0.003 μg/mL). The analysis of the metabolomics profile suggested that while P. variabilis extracts shared a common profile, the chemical profile of P. caribaeorum extracts was highly diverse across the different locations. This data pointed to the importance of marine microbiota in the production of secondary active metabolites within these specimens and suggest a preliminary correlation between bioactivity and microbial content. This discovery provides a key next step in developing tools to correlate metagenomic diversity of the bacteria associated with zoanthids to cytotoxicity and its associated bioactive secondary metabolites.

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Afternoon Session

OC 10

USING U SING THE THE P POLYTHEONAMIDE OLYTHEONAMIDE P PATHWAY ATHWAY TO E EX EXTEND TEND TH THE ES STRUCTURAL TRUCTU T R AL DIVERSITY DIVERSITY T OF OF RIBOSOMAL RIBOSOMAL PEPTIDES PEP E TIDES Brandon Brandon d I. I. Morinaka Mor o inaka1, Michael Michaeel F. Freeman Freeman1, Ch Christian ristian G Gurgui urggui2, Maximilian Maxi a milian J. Helf Helf1, Agu stinus R. U ria1, N Neil eil J. O Oldham lddham3, S Shigeki hige g kii M Matsunaga aattsunagga4, M Muriel uriel Gugger, Guggger,5 H Helge elgge B. Agustinus Uria 6 11,22 * 1, Bodee, an andd Jörn Jörn Piel Piel Bode, 1

In Institute stitute ooff M Microbiology, icrobiology, Eidgenössische Eidgenössische Technische Technische Hochschule Hochschule (ETH) (ETH) Zürich, Zürich, W Wolfgang-Pauli-Str. olffgang-Pauli-Str. 10, 10 8093 Zürich, Zürich, Switzerland. Switzerland.2 Ke Kekulé kulé Institute Institute fo forr O Organic rg a n ic C Chemistry hemistry and and Biochemistry, Biochemistry, University University of of Bonn, B onn , Ge rhard-Domagk Str. Str. 1, 1, 53111 53111 Bonn, Bonn, Germany. Germany.3 Sc School hool of of Chemistry, Chemistry, University University of of Nottingham, Nottingham, University U n iv e rs ity Gerhard-Domagk 4 Pa rk, Nottingham, N o ttin g h a m , N G72RD, UK. UK. Gr Graduate raduate School School of of Agricultural Agricultural and and Life Life Sciences, Sciences, The The University University of of Park, NG72RD, Tokyo, B unkyo-Ku, Tokyo Tokyo 113-8657, 113-8657, Japan. Japan.5 De Department partment of of Microbiology, Microbiology, Pasteur Pasteur IInstitute, nstitute, 2288 rrue ue dduu D Drr Tokyo, Bunkyo-Ku, Ro ux, 75015 75015 Paris, Paris, France. France.6 In Institute stitute ffor or M Molecular o le c u la r B Biosciences, iosciences, University University ooff F Frankfurt, rankfurt, Max-von-Laue-Str. Max-von-L Laue-Str. Roux, 9, 60 0438 Frankfurt Frankffuurt am am Main, Main, Germany. G e rm a n y . 60438 Em ail:[email protected], [email protected] [email protected] Email:[email protected], 17th September, 2013

U sing a metagenomic metagenomic approach appproach we we recently recently characterized charracterized the the biosynthetic biosynthetic pathway patthway of the the Using hi ghly cytotoxic, cytotoxic, sponge-derived sponge-deriveed polytheonamides. polytheonaamides.1 The The h pol polytheonamides, ytheonam mides, which whiich are are r highly produced produced by a bacterial bacterial symbiont, symbiont o , are arre ribosomally ribosomally synthesized syntthesized and and post-translationally post-trannslationally modified peptides (RiPPs). The gene onlyy ssix modifying modified pe ptides (Ri PPs ). T hee sstreamlined trreamlined ge ne ccluster luster ccontains ontains onl ix m o fyying odi enzymes responsible modifications. The precursor peptide enzymes re sponsible ffor oor 48 m o fications. T odi he lleader eader re rregion gion of tthe he pre cursor pe ptide resembles resembles members members of the the nitrile nitrile hydratase hydraattase family, faam mily, and annd is proposed p propos ed to to be characteristic chaarracteristic off a new (proteusins). demonstrated new family family of natural natuural products products (prot eusins). We We de monstraated tthe he ffunction unc u tion ffor or o aann unprecedented SAM unprecedented radical radical S AM epimerase epimerase (PoyD), (PoyD), which which generate gene n raate multiple multiple and annd different differe r nt Damino acid acid residues, residues, and annd likely likely carries carries out all all 18 epimerizations. epimerizattions. The The protein protein sequence sequeence of amino PoyD putative radical SAM genes genomes PoyD was was used used to to identify identify additional additiona o l put ative ra dical S AM eepimerase pimerase ge nes iinn tthe he ge enomes of cyanobacteria. cyaano n bacteria. The The genes genes were were located located in in putative putative proteusin prroteusin gene gene clusters clusters for for or as-yet as-yet unidentified unidentified natural naatural products. products. To To gain gain insight insight into into the the function fuuncction of these these enzymes, enzymes, the thee genes genes were were cloned clone o d into into E. E. coli coli and annd coexpressed coexpressed with with their their cognate cognaatte precursor precursor peptides. peptides. Chemical Chhemical analysis annalysis of the the modified modified precursors precursors o revealed revealed multiple multiple D-amino D-am mino acids, acids, with with each each epimerase epiimerase showing PoyD. Radical SAM represent robust new showingg different different selectivity selectivity tthan han P oyD. Ra dical S AM eepimerases p merases re pi present a robu ust ne w modifyying enzyme enzyme that thatt will will extend exttend the the function fuunction and annd structural structural diversity diversity of ribosomal ribos b omal modifying peptides.Figure peptides.Figure 1. Structure Structure off polytheonamides poolytheonaamides A aand nnd B.

References References 1. F Freeman, M. M. F., F., Gurgui, Gurgui, C., C., Helf, Helf, M. M. J., J., Uria, Uria, A. A. R., R., Oldham, Oldham, N. N. J., J., Matsunaga, Matsunaga, S., S., Sahl, Sahl, H.-G., H.-G., Piel, P ie l, Freeman, J Sc J. ience 2012, 338 87-390. Science 338,, 3387-390.


35


OC 11

Afternoon Session

A BACTERIAL SYMBIONT OF JAPANESE MARINE SPONGE Discordemia calyx, PRODUCES BIOLOGICALLY ACTIVE METABOLITES Toshiyuki Wakimoto1, Yoko Egami2, Yu Nakashima1, Ikuro Abe1 1

The University Of Tokyo, Graduate School Of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-Ku, 113-0033, Tokyo, Japan, 2The University Of Tokyo, Graduate School of Pharmaceutical Sciences, 7-3-1 Hongo, Bunkyo-Ku, 113-0033, Japan. E-mail: [email protected]


The lithistid sponges including the Discodermia and Theonella genera are exceptionally rich sources of structurally unique polyketides and peptides. Interestingly, certain classes of these metabolites were structurally similar to the metabolites of terrestrial bacteria, which have suggested the bacterial symbiont as the real producer of these sponge metabolites. However, the uncultivability of most symbionts has hampered the understanding of the producer organism, despite its potential to produce bioactive molecules. We first tried to obtain the biosynthetic gene cluster of two cytotoxic compounds, calyculins1 and calyxamides2 isolated from the Japanese sponge Discodermia calyx by a metagenome mining approach. With the desired PKS-NRPS hybrid gene clusters in hand, further attempts to identify the symbiotic bacteria having these two gene clusters were conducted by fluorescence in situ hybridization and laser microdissection for PCR analysis. As a result, both two gene clusters were encoded by same filamentous bacterium, which was identified as Entotheonella sp. based on the 16S rRNA sequence. Moreover, kasumigamide,3 a linear peptide originally isolated from a flesh water cyanobacterium, was also indentified from D. calyx. The single cell analyses of complex microbial assemblage revealed that kasumigamide is also biosynthesized by same Entotheonella sp. This is the solid proof of the long-suspected question regarding who the real producer of unique sponge metabolites is, demonstrating that there are indeed the talented but uncultured bacteria commonly residing in at least two genera, Discodermia and Theonella.

36



Afternoon Session

OC 12

GENOMIC INSIGHTS INTO THE DIVERSITY AND EVOLUTION OF SECONDARY METABOLITE GENES IN THE MARINE ACTINOMYCETE GENUS SALINISPORA Nadine Ziemer, Anna Lechner, Matthias Wietz, Krystle Chavarria, Natalie Millan, Paul Jensen Scripps Institution of Oceanography, Center For Marine Biotechnology And Biomedicine, 9500 Gilman Drive, La Jolla, 92093-0204, USA E-mail: [email protected]



The marine actinomycete genus Salinispora is a prolific producer of secondary metabolites including the anti-cancer compound salinosporamide A. A culture collection containing thousands of Salinispora strains derived from global collection sites provides a unique opportunity to address questions about the relationships between biogeography, secondary metabolite production, and species-level taxonomic assignments. In a massive sequencing project in collaboration with the Joint Genome Institute, we obtained genome sequences from 75 Salinispora strains isolated from seven different locations around the world. These genomes not only provide ample opportunities to guide our natural product discovery and engineering efforts, but the magnitude of the sequencing project provides unprecedented access into the evolutionary processes that create secondary metabolite structural diversity. We analyzed the genomes for the presence of secondary metabolite pathways using tools such as NaPDoS and antiSMASH and detected an enormous diversity of cryptic PKS and NRPS gene clusters. We present the diversity and distribution of these pathways in Salinispora species and give a first insight into the mechanisms that drive pathway evolution and the existence of “genomic hot spots” for secondary metabolism.

37


IL 6

Afternoon Session

INVITED LECTURE 6

DIDEMNINS AND THALASSOSPIRAMIDES—GIFTS AWARDED BY GENOME MINING OF Tistrella Pei-Yuan Qian Global Collaborative Research Program of KAUST, Division of Life Science, Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong. E-mail: [email protected]


The anti-neoplastic agent didemnin B from the Caribbean tunicate Trididemnum solidum was the first marine drug that was clinically tested in humans. Because of its limited supply and complex cyclic depsipeptide structure, considerable challenges were encountered during the development didemnin B as commercial drug, leading to development of aplidine (dehydrodidemnin B) that is currently being evaluated in numerous clinical trials. Recently, we found that the marine proteobacteria Tistrella mobilis and Tistrella bauzanensis could produce didemnin B and its derivatives through a unique post-assembly line maturation process. Analysis of complete genome sequence of the 6,513,401 bp T. mobilis strain KA081020-065 with its five circular replicons revealed the didemnin biosynthetic gene cluster (DID) on the 1,126,962 bp megaplasmid pTM3. The DID locus encodes a 13-module hybrid non-ribosomal peptide synthetase-polyketide synthase enzyme complex organized in a collinear arrangement for the synthesis of the fatty acylglutamine ester derivatives didemnins X and Y rather than didemnin B as first anticipated. The discovery of the didemnin biosynthetic gene cluster may provide a long-term solution to the supply problem that presently hinders drug development of this group of marine natural products and also pave the way for the genetic engineering of new didemnin congeners.Further genome mining of two species of Tistrella revealed another NRPS-PKS hybrid gene cluster encoding unidentified products. Successful culture medium manipulation led to the high expression of the gene cluster and discovery of as many as 14 new thalassospiramide derivatives in addition to the isolation of two known compounds, thalassospiramides A and B. The bioinformatics analysis of the gene clusters suggests that structural diversity, which ranges from the 803.4 Da thalassospiramide C to the 1291.7 Da thalassospiramide F, were resulted from a complex sequence of reactions involving amino acid substrate channeling and enzymatic multimodule skipping and iteration. Preliminary biochemical analysis supported a biosynthetic model in which in cis amino acid activation competed with in trans activation to increase the range of amino acid substrates incorporated at the N terminus. These compounds represent a new class of calpain 1 protease inhibitors with nanomolar activity and low toxicity. Further chemical modification of these compounds and docking experiments between them and calpain suggested that the , -unsaturated keto is a key functional group that targets the active cysteine residue.

Acknowledgement This work was carried out by Dr. Ying XU, Mr. Liang LU from PY Qian’s lab at Hong Kong University of Science and Technology, and by Mr. Roland Kersten and Dr. Avena Rose In B Moore’s lab at Scripps Institution of Oceanography, UCSD; W Fenical assisted with structure identification of Didemnin B; Funding support was from Chinese Ocean Mineral Resource Research and Development Association and Global Collaborative Research of KAUST to PY Qian, and NIH grants to BS Moore, W Fenical and PC Dorrestein.

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Thursday, 19th September, 2013

Unlocking the Potential of Marine Organisms Unlocking the Potential of Marine Organisms MORNING SESSION: Frontiers of Discovery & Innovation Chair: Junichi Tanaka 09:00 - 09:40 PL 4 Tadeusz Molinski University of California, San Diego, USA 09:40 - 10:10 IL 7 M. Valeria D'Auria University of Naples "Federico II", Italy 10:10 - 10:30 OC 13 Ronald J. Quinn Eskitis Institute, Griffith University, Australia 10:30 - 10:50 OC 14 Mark Hamann University of Mississippi, USA 10:50 - 11:30 Coffee break 11:30 - 11:50 OC 15 Matthias Köck Alfred-Wegener-Institut, Germany 11:50 - 12:10 OC 16 Gabriele Koenig University Bonn, Germany 12:10 - 12:30 OC 17 William H. Gerwick University of California San Diego, USA 12:30 - 12:50 OC 18 Philipp Hess Ifremer, Laboratoire Phycotoxines, France 12:50 - 13:20 IL 8 Shigeki Matsunaga The University of Tokyo, Japan 13:20 - 14:30 Lunch AFTERNOON SESSION: Marine Ecology: Understanding Marine Organisms Chair: Raymond Andersen Patrizia Ciminiello University of Naples "Federico II", Italy 14:30 - 15:10 PL 5 Vassilios Roussis University of Athens, Greece 15:10 - 15:40 IL 9 Marie-Lise Bourguet-Kondracki UMR 7245 CNRS-MNHN, France 15:40 - 16:00 OC 19 Miguel Costa Leal Universidade de Aveiro, Portugal 16:00 - 16:20 OC 20 Coffee break 16:20 - 17:00


39


PL 4

Morning Session

PLENARY PL ENARY LECTURE LECTURE 4

HETEROCYCLIC FROM MARINE SPONGES: SYNTHESIS HETE EROCYCLIC COMPOUNDS COMPOUNDS F ROM M A RIN E S PONGES: S YNTHESIS IIN N AID AID OF OF STRUCTURE STRUCTURE ELUCIDATION ELUCIDATION AND AND SAR SAR Tadeusz T ade a usz (‘Ted’) (‘T Ted’) M Molinski oollinskii Department De partment of of Chemistry Chemistry and and Biochemistry, Biochemistry, Un University iversity ooff California, C a lif o r n ia , S San an Diego Diego Gilman Dr. Dr. MC0358, MC0358, La La Jolla, Joolla, CA CA 92093, 922093, USA. USSA. 9500 Gilman E-mail: tm olinskicsd.edu E-mail: tmolinskicsd.edu


The marine natural products (MNPs) The wide wide d chemical chemical diversity diversity that thaat characterizes chaarracterizes tthe he m arrinee nat tural produc ts (M NPs) from from r invertebrates, invertebra b tes, particularly paarrticularl r y marine mari r ne sponges, sponges, is is often offten matched matched by potent potent biological biological activity activity in in therappeut u ically re levannt bi oassayyss. Several Several marine-derived marrine-derived natural naatural products products have haave v now noow been been therapeutically relevant bioassays. approved appproveed as as drugs – one from frrom a cone-snail cone-snail (Prialt™) (Prialt™) and annd one from frrom a marine marrine tunicate tunicate (Yondelis™) – while while others others have havee iinspired nspired ‘de rived-drugs’.1 In this thhis talk, talk, I will will attempt attempt to to (Yondelis™) ‘derived-drugs’. illustratte aspects aspects of recent recent research researcch on marine marrine natural nattural products prodducts from from r our labs labs at at UCSD, UCS C D, La La illustrate Jolla, Jolla, California.Case C lifoornia.Case studies Ca studies will will be presented presented that thatt elucidateextraordinary eluc u idatteextraordinary heterocyclic heteroc r yclic moleculeswith moleculleswith significant siggnificant biological biologggical activitiesusing activitiesusingg ‘nanomole-scale’ ‘nannoomole-scale’ methodology, methodology gyyy, total total synthesis, origin (PIAs) using synthesis, and and n exploration exploration of the thee ori gin of pyrrole-imidazole pyrrole-imidazole aalkaloids lkaloids (P IAs) us ing n ccellellfree fre r e labeling labbeeling studies stuudies – so-called so-called ‘metabiosynthesis’. ‘metabbiosynthesis’. A common com mmon theme theme merges merges that thaat unifies unifies these sstudies tudi u es aand nnd unde rscores ourr aapproach:expanded ppproach:expannded sscope coppe of di scoverythrough ssynthesisynnthesisthese underscores discoverythrough 3 enabled enaabbled structure structure elucidation elucidattion aand nnd sstructure-activity tructure-activity re relationship lattionnship sstudies tudies (S (SAR). AR).

References R efeerences M lin s k i, T Mo alisay, D. D. S.; S.; Lievens, Lievens, S. S. L.; L .; S aludes, JJ.. PP.. ""Drug D ru g D evelopment from from Marine Marine 1. Molinski, T.. F F.;.; D Dalisay, Saludes, Development N tural Products." Na P r o d u c ts ." Natural N t. R Na ev. D rug Discov.2009, Discov.2009, 8,, 669-85. 9-85. Nat. Rev. Drug ( ) Stout, (a Stout, E. E. P.; P.; Wang, Wang, Y-G.; Y-G.; Romo, Romo, D.; D.; Molinski, Molinski, T. T. F. F. Angew. Angew. Chem. Chem. Intl. Intl. Ed. Ed. 2012, 51, 51, 44877. 877. (b) (b ) 2. (a) S out, E. St E. P.; P.; Morinaka, Morinaka, B. B. I.; I.; Wang, Wang, Y-G.;Romo, Y-G.;Romo, D.; D.; Molinski, Molinski, T. T. F. F. J. N a t. P rod.2012, 75, 5527 27 Stout, Nat. Prod.2012, M lin s k i, T Mo .F.; M o rin a k a , B Integrated Approaches Approaches to to the the Configurational Configurational Assignment Assignment of of Marine M a rin e 3. Molinski, T.F.; Morinaka, B.. II.. ““Integrated N tu r a l P Na roducts” Tetrahedron2012, Tetrahedron2012, 2 68, 99307-9343 307-9343 Natural Products” 68,

40



Morning Session

IL 7

INVITED LECTURE 7

MARINE COMPOUNDS AS MODULATORS OF METABOLIC NUCLEAR RECEPTORS: ISOLATION, DESIGN, TOTAL SYNTHESIS AND POTENTIAL THERAPEUTIC APPLICATION Maria Valeria D’Auria1, Valentina Sepe1, Carmen Festa1, Simona De Marino1, Raffaella Ummarino1, Giuseppe Bifulco2, Barbara Renga2, Stefano Fiorucci2, and Angela Zampella1 1

Dipartimento di Farmacia, Università di Napoli “Federico II”. 2Dipartimento di Farmacia, Università di Salerno, Fisciano (SA). 3Dipartimento di Medicina Clinica e Sperimentale, Università di Perugia, S. Andrea delle Fratte, Perugia E-mail: [email protected]


Nuclear receptors (NRs) are members of a superfamily of ligand-dependent transcription factors that regulate diverse aspects of reproduction, development, homeostasis and immune function. Among them, FXR, PXR and PPAR , belonging to the metabolic receptor subfamily of NRs, participate in an intricate network of interactions that ultimately govern lipid, steroid, and glucose homeostasis and represent a crossroad between metabolism, inflammation and cancer. Therefore, new molecular entities that modulate the activity of these NRs are considered very attractive targets for treatment of inflammation-driven immune dysfunctions, metabolic diseases and cancer. Recent studies by our group disclosed for the first time several marine metabolites as modulators of metabolic receptors. Our investigation led to the identification of some lead compounds with peculiar activation profile, such as solomonsterols, potent agonists for PXR; theonellasterol and suvanine, selective antagonists of FXR; plakilactone C and gracilioethers, covalent or not-covalent ligands of PPAR . The study has involved structural identification of new marine modulators of NRs, in vitro and in vivo pharmacological investigation, detailed analysis of the molecular basis of ligand-protein interaction, experimental validation of the ligand-receptor binding, medicinal chemistry lead optimization by semi-synthetic or synthetic modification of the natural scaffolds. Significant results of this multi-parametric study will be discussed in the present communication.

References 1.

Fiorucci, S.; Distrutti, E.; Bifulco, G.; D'Auria, M.V.; Zampella, A. Trends in Pharmacological Sciences, 2012, 33 , 591-601


41


OC 13

Morning Session


COMPUTATIONAL METHODS IN MARINE NATURAL PRODUCTS AND FINGERPRINTING THE DRUG-LIKE NATURAL PRODUCT METABOLOME Ronald J Quinn Eskitis Institute, Griffith University, Brisbane, Queensland 4111, Australia E-mail: [email protected]

The revival of natural products arises because of their novel chemotypes and because of the inherent difficulty associated with producing synthetic libraries that contain molecules that interact with biology space. Natural products have an inherent understanding of biology space. Our lead discovery program is based on the drug-like natural product metabolome.1,2 Natural products and their analogues have had high impact as drugs because of the embedded biosynthetic molecular recognition that transfers to therapeutic targets as described by protein fold topology (PFT).3,4 We report the use of computational methods that may be used to guide marine natural product research: drug-like and lead-like properties 19th September, 2013

natural product scaffold properties using Fsp3 to evaluate structural complexity natural products as lead-structures; chemical transformations to create leadlike libraries. approaches to automate protein fold topology interrogation of large databases. We have previously reported a strategy (requiring no knowledge of structure) for the generation of lead-like enhanced (LLE) extracts and fractions with a protocol that allowed the retention of lead- and drug-like constituents by selecting favourable physicochemical properties such as log P < 5. We report on the use of the LLE fraction library protocol to create proton nuclear magnetic resonance-based metabolic fingerprint of the LLE fractions and introduce structural information early in the natural product isolation workflow. This approach is illustrated using a sponge Iotrochota sp., where several interesting metabolites were identified on small-scale and the subsequent large-scale extraction work yielded new natural products iotrochotadines A –D, and a novel compound iotrochotazine A. References

42

1.

Camp, D.; Davis, R. A.; Campitelli, M.; Ebdon, J.; Quinn, R. J. Drug-like properties: guiding principles for the design of natural product libraries. J. Nat. Prod. 2012, 75, 72-81.

2.

Camp, D.; Davis, R. A.; Evans-Illidge, E. A.; Quinn, R. J. Guiding principles for natural product drug discovery. Future Med. Chem. 2012, 4, 1067–1084.

3.

McArdle, B. M.; Campitelli, M. R.; Quinn, R. J. A common Protein Fold Topology shared by flavonoid biosynthetic enzymes and therapeutic targets. J. Nat. Prod. 2006, 69, 14-17.

4.

Kellenberger, E.; Hofmann, A.; Quinn, R. J. Similar interactions of natural products with biosynthetic enzymes and therapeutic targets could explain why nature produces such a large proportion of existing drugs. Nat. Prod. Rep. 2011, 28, 1483-1492.



Morning Session

OC 14

USING HOST-ASSOCIATED BACTERIA AND SYNTHESIS TO CONSTRUCT COMPLEX AND NOVEL MARINE CYTOTOXINS WITH UNIQUE MECHANISMS OF ACTION Mark T. Hamann Department of Pharmacognosy, Pharmacology, Chemistry and Biochemistry, The University of Mississippi. 407 Faser Hall, 38677, Oxford, Estados Unidos E-mail: [email protected]

In this presentation we will review progress on the discovery and development of new marine NP prototypes for the control of cancer. The kahalalides are dual targeted NPs that act on HER-3 over-expressing cells that perturb the membrane of the lysosome. Two leads from this class have advanced to clinical trials by PharmaMar however challenges with half-life and efficacy continue to be a limitation. Second-generation analogs of this class of TM



lipopeptide take into consideration improvements to the lipopeptide Caspofungin completed by investigators at Merck as well as production of the NP from the hostassociated Vibrio sp. The karlotoxins are a group of membrane interacting agents that will form a hairpin around cholesterol. Details of the MOA remain to be determined but impressive selectivity for NSCLC over nerve cell carcinoma has resulted in a promising lead without the typical dose limiting toxicity to nerve cells carcinoma has resulted in a promising lead without the typical dose limiting toxicity to nerve cells in vitro for many cancer drugs. A total synthesis of the KmTx class is nearly complete. From the deep surveys of the Aleutian Islands two new classes of alkaloids have been identified. One of these new class of alkaloids targets pancreatic cancer and a second targets melanoma.

43


OC 15

Morning Session


NEW CYSTOCHROMANES AND OXIDIZED MERODITERPENES FROM THE BROWN ALGA Cystoseira baccata. INDICATION FOR A GENERAL REVISION OF THE BICYCLO[4.3.0]NONANE SYSTEM?

Julie Muñoz1, Alexis Krupp2, Stefan Immel2, Michael Reggelin2, Gerald Culioli3, and Matthias Köck1,* 1

Alfred-Wegener-Institut, Helmholtz-Zentrum für Polar- und Meeresforschung, Am Handelshafen 12, 27570 Bremerhaven, Germany, 2Clemens-Schöpf-Institut für Organische Chemie und Biochemie, Technische Universität Darmstadt, Petersenstraße 22, 64287 Darmstadt, Germany, 3Université de Toulon, MAPIEM, EA 4323, 83957 La Garde cedex, France. E-mail: [email protected]


Cystoseira is one of the most studied genus of the Sargassaceae family and Cystoseira spp. are known to produce a wide array of terpenes, such as linear diterpenes or meroditerpenes [1-3]. Even though these compounds have been studied for more than 40 years, it was recently demonstrated that their structures could still raise interesting issues, especially concerning their stereochemistry [4,5]. The isolation of several new meroditerpenes from the brown alga C. baccata led to a strong indication for a general revision of the bicyclo[4.3.0]nonane system for compounds of this family. A detailed study of the relative and absolute configurations of the new molecules was carried out. For the determination of the relative configuration, NOE-derived interproton distances were used as input for floating chirality DG/DDD simulations. Since the number of NOEs was not sufficient for an unambiguous assignment of the relative configuration, residual dipolar couplings (RDC) were used to refine the structures. The absolute configuration of the new compounds was assessed by comparison of the circular dichroism (CD) spectra to the calculated spectra. References 1. 2. 3. 4.

5.

44

Blunt, J. W.; Copp, B. R.; Hu, W.-P.; Munro, M. H. G.; Northcote, P. T.; Prinsep, M. R. Marine natural products. Nat. Prod. Rep., 2008, 25, 35-94. Valls, R.; Piovetti, L. The chemistry of the Cystoseiraceae (Fucales: Pheophyceae): chemotaxonomic relationships. Biochem. Syst. Ecol., 1995, 23, 723-745. Amico, V. Marine brown algae of family Cystoseiraceae: chemistry and chemotaxonomy. Phytochemistry, 1995, 39, 1257-1279. Mokrini, R.; Ben Mesaoud, M.; Daoudi, M.; Hellio, C.; Maréchal, J.-P.; El Hattab, M.; OrtaloMagné, A.; Piovetti, L.; Culioli, G. Meroditerpenoids and derivatives from the brown alga Cystoseira baccata and their antifouling properties. J. Nat. Prod., 2008, 71, 1806-1811. Muñoz, J.; Culioli, G.; Köck, M. Isolation and reassignement of the configuration of meroditerpenes from the brown alga Cystoseira baccata, manuscript in preparation.



Morning Session

OC 16

ORAL OR AL C COMMUNICATION OMMUNICATION N 16 16

SALIMABROMIDE: UNEXPECTED UNEXPEC E TED CHEMISTRY CHEMISTR RY FROM FROM THE THE OBLIGATE OBLIGATE E SALIMABROMIDE: MARINE MYXOBACTERIUM MYXOBA B CTERIUM Enhygromyxa Enhyggrom myxa x Salina Sallina MARINE Gabriele M. M. König, Königg,, S Stephan teph p an Felder, F lde Fe d r, S Sandra andra Dreisigacker, Dreisigac g ke k r, Stefan Steffan Kehraus, Kehraus, Edith Ediith Gabriele Neu, Gabriele Gabriele Bierbaum, Bierbaum, Patrick Pat atrick R. W rigght, D irk M enche, T ill F. S chäberle Neu, Wright, Dirk Menche, Till Schäberle In Institute stituute ffor or P Pharmaceutical harmaceutical Biology, Biology, Ke kulé-Institute ffor or O rganic C hemistry aand nd Biochemistry, Bioc o hemistry, Kekulé-Institute Organic Chemistry In stitute ooff M e d ic a l M icrobiology, Immunology Immunology and and Parasitology, Parasitology, University University of of Bonn Bonn Institute Medical Microbiology, E-m mail: [email protected] [email protected] E-mail:


T hee ffocus oc o us u of o the thee current cuurreent study study is is on o unusual, uunus usual, hitherto hitheerto not ot at at all all researched reesearrccheed bacterial bacterial taxa. taxa. The W ere r successful successful u in in isolating isolatingg an an obligatory obligaatory marine marrine myxobacterium myxobacterium from from r mud Wee w were a mud ssample ample ffrom rrom a ccoastal oastal ar rea of tthe he iisland sland P rerow (G erm r anny). 16S rD NA an nalysis area Prerow (Germany). rDNA analysis re vealed tthis hhis bacterium bacterium aass Enhygromyxa Enhyyggrom myxa ssalina, alina, a sspecies pecies closely closely related related to to revealed m icroorrgaannisms to to date datte termed termed as as “unculturable”. “unculturabble”. T The he pe persistent rsistent antibiotic anntibiotic aactivity ctivity microorganisms ttowards owarrds Gram-positive Graam-positive microorganisms microorggannisms let let us choose choose this this bacterium bacterium for for o detailed detailed cchemical hemical iinvestigation. nvestigaation. L arrge sscale cale ccultivation ultivaation m ade iitt ppossible ossible ttoo iisolate solate the the first first secondary secondaarry metabolite, metabbolite, i.e. i.ee. 1, Large made marine myxobacterium. The only produced minute ffrom rom r tthis hiis m arrine m yxobacterium. T he ccompound ompound iiss onl y pr roduced iin nm inute aamounts mountts of μg/L. hass a novel 8μ g/L. Salimabromide Salimabbromide ((1) 1) ha novel carbon carrbon skeleton, skeleton, on and annd the thhe core core structure structure reveals reveals no resemblance resemblannce to to any any known know wn natural naatural or synthetic synthetic compound. compound. The The absolute abs b ol o ute cconfiguration onffigurration was was deduced deduced using usingg experimental experimental and annd calculated calcullated CD spectroscopic spectroscopic data. data. da Further novell ccompounds were and will presented. F urther nove ompounds w ere aalso lso ffound ound o an nd ttheir heir sstructures trructures w ill be pre sent n ed. Phylogenetic analysis draft genome data E.. ssalina SWB007, P hylogeenetic an nalysis aand nnd ffirst irst dra aft ge nome da ata ffrom rom r E alina sstrain train S WB007, llead ead us ttoo cconclude onclude tthat haat m marine arrine m myxobacteria yxobaacteria ar are re phyl phylogenetically ogenetically qui quite te di distinct stinct ffrom rom r other other myxobacteria. This m yxobaacteria. T his iiss also also mirrored mirroreed in in their their biosynthetic biosyntthhetic abilities. abi b lities.

Figure. Novel Fi iggure. N ovel ttricyclic ricyclic ccarbon arbon sskeleton keleton and annd structure structure u off salimabromide salimabromide


45


OC 17

Morning Session


BASTIMOLIDE A, A BIOSYNTHETICALLY INTRIGUING POLYHYDROXY MACROLIDE WITH POTENT ANTIMALARIAL ACTIVITY William H. Gerwick1,6, Chang-Lun Shao1,2, Roger G. Linington3,4, Marcy J. Balunas4,5, Paul Boudreau1, Tara Byrum1, Charlie Larson1, Lena Gerwick1, Niclas Engene7 and Chang-Yun Wang2 1

Center for Marine Biotechnology and Biomedicine, Scripps Institution of Oceanography, University of California San Diego, La Jolla, California 92093, USA. 2Key Laboratory of Marine Drugs, The Ministry of Education of China, School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, P.R. China. 3Department of Chemistry and Biochemistry, University of California Santa Cruz, Santa Cruz, California 95064, USA. 4Instituto de Investigaciones Científicas y Servicios de Alta Tecnología, Clayton, Apartado 0816-02852 Panama. 5School of Pharmacy, University of Connecticut, Storrs, CT 06269, USA. 6 Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, La Jolla, California 92093, USA. 7Smithsonian Marine Station, 701 Seaway Dr., Fort Pierce, FL 34946 E-mail: [email protected]


Bastimolide A, a biosynthetically intriguing class of polyhydroxy macrolide possessing a 40-membered ring, was isolated from a new genus of the cyanobacterium Tropicimarinus villus collected from the Caribbean coast of Panama. The planar structure of this new polyketide macrolide is composed nine hydroxy groups, one ester, a trisubstituted olefin and one unique t-butyl group, and was elucidated by a combination of spectroscopic data and chemical reactions. Its complete structure and absolute configuration were unambiguously determined by X-ray diffraction analysis of the nona-p-nitrobenzoate derivative. Bastimolide A showed potent antimalarial activity against four resistant strains of Plasmodium falciparum with IC50 values between 80 and 270 nM, and thus represents an exceptional new structural class of promising antimalarial lead compound.

46



Morning Session

OC 18


HIGH RESOLUTION MASS SPECTROMETRY AND BIOLOGICAL TOOLS FOR SCREENING OF MARINE BIOTOXINS AND DISCOVERY OF MARINE NATURAL PRODUCTS VIA DEREPLICATION Philipp Hess1, Bernhard Wüst2, Thomas Glauner2, Yann Guitton1, 3, Florence Mondeguer1, Manoëlla Sibat1, Zita Zendong1, 3, Véronique Séchet1, Thierry Jauffrais3, Marie Geiger1, 3, Christine Herrenknecht3 and Zouher Amzil1 1

Ifremer, Laboratoire Phycotoxines, Rue de l’Iled’Yeu, 44311 Nantes Cedex 03, France. 2 Agilent Technologies, Hewlett-Packard-Str. 8, 76337 Waldbronn, Germany. 3 Université de Nantes, Faculté de Pharmacie, MMS EA 2160, 44000 Nantes, France. E-mail: [email protected]

Marine natural products are unsurpassed sources of chemical diversity underlying the natural biodiversity. Libraries have been established and contain a large number of compounds, e.g. MarinLit™ (22,000 compounds) or AntiMarin™ (ca. 50,000 compounds). This paper describes the use of high resolution mass spectrometry and miniaturised bioassays for the dereplication of marine natural products. 19th September, 2013

For full scan screening and dereplicationof algal and shellfish samples, a generic gradient method allowed for molecular feature extraction which yielded typically several hundred compounds for micro-algal fingerprints of single species. An in-house database was developed to have an authoritative list of ca. 300 known marine toxins.Screening of this in-house database and library facilitated rapid and definitive toxin identification in algal and shellfish samples, while comparison to a large-scale database (ca. 30,000compounds) resulted in tentative identification of ca. 10 % of all compounds present at 1 ppm mass accuracy (initial dereplication in ca. 15 min/sample). Statistical approaches using Mass Profiler Professional software reduced data complexity and streamlined the dereplication workflow. Cytotoxicity and fly larval toxicity assays applied to organisms of several groups including cryptophytes, mammiellophytes, prymnesiophytes and dinoflagellates, pointed to 4 toxic dinoflagellates (Azadinium spp., Prorocentrum lima, Alexandrium ostenfeldii and Vulcanodinium rugosum). Analysis using the above-mentioned full scan mode techniques allowed for qualitative and semi-quantitative profiling of these dinoflagellates. Dereplication allowed for rapid preliminary information regards toxic components. Overall, the study demonstrates the power of high-resolution mass spectrometry as part of a toxicity directed analysis approach for the screening of bioactive natural products.


47


IL 8

Morning Session

INVITED LECTURE 8

MARINE NATURAL PRODUCTS WITH ANTI-CANCER RELATED ACTIVITIES Shigeki Matsunaga, Kentaro Takada, Yuki Hitora, Akihiro Ninomiya, Yasufumi Imae, Yi Sun, Masato Naruse, Emi Takanashi Laboratory of Aquatic Natural Products Chemistry. Graduate School of Agricultural and Life Sciences. The University of Tokyo. 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657.Japan. E-mail: [email protected]

In our continuing study toward the discovery of anticancer leads, we have searched for compounds through cell-based and enzyme inhibitory assays, which are all related to carcinogenesis. The isolation, structure elucidation, and biological activities of several


marine natural products will be discussed.

48



Afternoon Session

PL 5

PLENARY LECTURE 5

MEDITERRANEAN HARMFUL ALGAL BLOOM EVENTS: OLD AND NEW RISKS Patrizia Ciminiello1, Carmela Dell"Aversano2, Martino Forino2, Luciana Tartaglione2 2

1 Department of Pharmacy, Via Domenico Montesano, 49, 80132, Napoli, Italy. UNIVERSITY OF NAPOLI "FEDERICO II", Department Of Pharmacy, Via D. Montesano, 49, Napoli, 80131, Italy. E-Mail: [email protected]


Seawater teems with phytoplankton species. Most of them are harmless; but, a few dozens, classified as toxic, under appropriate climatic and environmental conditions can bloom to such an extent to give rise to exceptional events, defined Harmful Algal Blooms (HABs). HABs exert their adverse impacts on both the environment and living organisms through the production of marine biotoxins. Such toxins are the causative agents of serious, sometimes even lethal, human intoxications. In the late ‘80s we afforded the first experimental evidence over the occurrence of a marine biotoxin in Italy, by isolating and characterizing okadaic acid (OA). Since then, our study has been a cornerstone in analyzing algal toxic outbreaks along the Italian coastline and has shown that the HABs along these coasts are in continuous evolution, with an increasing number of toxic compounds belonging to different classes of toxins1. The latest threat to public health is placed by some Ostreopsis spp. - mostly O. cf ovata and O. cf. siamensis, that have become increasingly frequent in the last decade. Liquid chromatography-mass spectrometry (LC-MS) analysis revealed that O. cf. ovata produces palytoxin-like compounds, ovatoxins, a class of very potent marine toxins. On the contrary, O. cf. siamensis was found to be practically devoid of any appreciable toxicity. LC-MS also played a key role for determining the accumulation of ovatoxins in both mussels and marine aerosols, as well as the influence of several environmental parameters on O. cf. ovata toxin production. References 1. 2.

D. M. Anderson (1989) In: Red tides: Biology Environmental Science and Toxicology (Eds.: T. Okaichi, D. M. Anderson, T. Nemoto), Elsevier, New York, pp. 11–16 P. Ciminiello et al. (2012) In: Handbook of Marine Natural Products (Eds.: E. Fattorusso, W.H. Gerwick, O.Taglialatela-Scafati) Springer Science+ Business Media B.V., pp. 1345-1387


49


IL 9

Afternoon Session

INVITED LECTURE 9

CHEMICAL ECOLOGY FOR ECOLOGICAL CHEMICALS Vassilios Roussis University of Athens, School of Pharmacy, Department of Pharmacognosy and Chemistry of Natural Products, Panepistimiopolis Zografou, Athens 15771, Greece. E-mail: [email protected]


The biodiversity of the Mediterranean basin ecosystem has been proven a prolific source of bioactive natural products. Interest in the chemical ecology of marine organisms has grown over the past decades focusing on the elucidation of the ecological functions of secondary metabolites. Biofouling is one of the most significant problems that marine organisms are facing. The same phenomenon is taking place on any surface immersed in sea water thus creating serious problems in the maritime domain. After the prohibition of TBT containing paints, there is an urgent need for the development of new environmentally benign antifouling alternatives that would be efficient against the most severe fouling organisms. In the framework of our search towards the isolation of metabolites with antifouling activity we have investigated a number of epibiont-free organisms collected from the Greek seas. Extracts and metabolites of the red algae genera Laurencia and Sphaerococcus exhibited significant inhibition on the settlement of barnacle cyprids. Bromosphaerol was found to be among the most active compounds exhibiting the lowest EC50 without toxic effects on the barnacle cyprids. The search for even more active analogues led to the isolation of a series of new minor brominated diterpenes. Targeting the development of a novel, cost-efficient, eco-friendly antifouling system we designed and tested experimental paints incorporating various amounts of copper oxide nanocontainers loaded with Laurencia and Sphaerococcus metabolites. The promising results might pave the way towards a sustainable solution for the persisting and expensive problem of biofouling. Acknowledgements This work is supported by the project “MARIPAINTS”, which is implemented under the "COOPERATION 2011" Action of the Operational Programme “COMPETITIVENESS AND ENTREPRENEURSHIP” and is co-funded by the European Social Fund (ESF) and National Resources.

50



Afternoon Session

OC 19


EXPLORING THE MICROBIAL DIVERSITY ASSOCIATED WITH A CARNIVOROUS SPONGE Samuel Dupont1, Erwan Corre2, Alyssa Carré-Mlouka1, Yanyan Li1, Isabelle DomartCoulon1, Jean Vacelet3 and Marie-Lise Bourguet-Kondracki1 1 Laboratoire Molécules de Communication et Adaptation des Micro-organismes, UMR 7245 CNRS, Muséum National d’Histoire Naturelle, 75005, Paris, France, 2 CNRS, UPMC, FR2424, ABiMS, Station Biologique, 29680, Roscoff, France, 3 Université Aix Marseille, CNRS, IMBE UMR 7263, 13007, Marseille, France E-mail: [email protected]

The microbial community associated with the carnivorous Demosponge Asbestopluma hypogea (Cladorhizidae family) was investigated through a multidisciplinary study. This study was stimulated by the discovery of A. hypogea in 1996 in a cave of the Mediterranean coast at 20 meters deep,1 while carnivorous sponges are usually found in deep-sea environments. Preliminary electron microscopy studies had shown that this tiny sponge, which is not a filter-feeding sponge and has no digestive cavity, harbors a highly abundant microbial community in its mesohyl. The increase in microbial density around the preys after their capture suggested a bacterial involvement in digestion.2 19th September, 2013

Exploring the microbial community associated with this carnivorous sponge, multidisciplinary approaches have been developed. They constitute a vast study with transmission electron microscopy observations and fluorescence in situ hybridization experiments in order to localize the bacteria and archaea in the sponge tissues. A metagenomic study using 454 pyrosequencing allowed, for the first time, the characterization of the associated microbial community targeting the V3 and V6 hypervariable regions of the 16S rRNA gene. We compared one A. hypogea sample from the cave environment to one sample maintained one year in an aquarium. Analysis of more than 60,000 sequences showed the presence of 20 bacterial phyla and 2 archeal phyla (38-54 % of sequences).3 Furthermore, the cultivable bacteria isolated on various media were identified by comparison of the 16S rRNA gene sequences. Evaluation of their antimicrobial, antioxidant and chitinolytic activities will also be presented and discussed. References 1. 2. 3.

Vacelet J & Duport E (2004), Zoomorphology 123: 179-190. Vacelet J, Boury-Esnault, N. (1996), Bulletin de l'institut royal de sciences naturelles de Belgique 66: 109-115. Dupont S, Corre E, Li Y, Vacelet J, Bourguet-Kondracki M-L, FEMS Microbiol. Ecol. (2013), accepted.


51


OC 20

Afternoon Session

BIOGEOGRAPHY MARINE NATURAL PRODUCTS RESEARCH BI IOGEOGRAPHY IIN NM AR INE N ATURAL P RODUCTS R ESEARCH Miguel Costa Leal M igguel Cos ta L eall1, M Murray urray M Munro unro2, Joh John n Bl Blunt unt2, João Puga Puga g 3, Car Carolina ollina M Madeira ade a ira4, 5,6 5,6 4 1 Bruno Jesus Jesus , Ru Ruii Rosa Rosa an andd Ricardo Ricardoo Calado Callaado Bruno 1

Depa Departamento part r amento de Bi Biologia ologia & CE CESAM, SAM , U Universidade niversidade dee A Aveiro, veiro, A Aveiro, veiro, P Portugal. ortugal. Department D epa parrtment off Che Chemistry, mistry, U University n versity off Ca ni Canterbury, annterbury, C Chri Christchurch, stchurch, N New ew Z Zealand ealandd 3 Zoo Z ooo Logical, Logical, L Lisboa, isboa, Portugal. Portugaal.4 Ce Centro ntro de Oceanografia, Oceannografia, Faculdade Faculdade de Ciências, Ciências, Universidade de L isboa, L isboa, P ortugal. 5

Universidade Lisboa, Lisboa, Portugal. ! " ! " # $ # $ 6 % &'()* &+$, % &'()* &+ $ , - L LUNAM UNAM université, université, U Université niversité dee Nant n es, Nantes, Nanntes, France Fraanc n e Nantes, E-maail:[email protected] m E-mail:[email protected] 2


Biogeoggraapphy aims aims to to describe describe and annd understand understand the the spatial spaatial patterns patterns of biodiversity. biodiverssity. Biogeography While While it it has has been been traditionally traditionally associated associaatted with with biology biology and anndd macroecology, macroecology, it it has has seldom seldom d been applied apppplied to to marine marrine natural natuural products prroducts research. researrch. Information Infform o ation on biodiversity biodiversity hotspots, hotspot p s, been sppecies distribution distrribution and annd latitudinal/longitudinal latitudinal/longi g tudinal gradients ggradients may may be useful useful u to to understand understand n species and annd predict prredict the the distribution distribution of marine marine natural naatural products. prroducts. Furthermore, Furthermore, such such biogeoggrapphical analysis analysis contributes contribuutes to to better better understand understannd spatial spatial and and n temporal temporal patterns paatterns biogeographical in the the chemical chemical composition composition of particular parrticularr biological pa biological groups grouups from from r different regions, regionss, as as in different well as to to identify identiffyy unscreened unscreened taxa taxa x and annd locations. locations. All All of these theese features features help help to to maximize maximize the the discovery discove o ry off marine marine natural naatural products. prroducts. The lack lack of application appplicattion of biogeography bioggeograapphy to to marine marine natural naatural products products research researc r h is is a The consequuence of the the lack lack of a comprehensive com mprehensive dataset dataset containing containing chemical, chemical, taxonomical taxonom mical consequence annd, nd most mos o t importantly, importantly, y geographical geograapphhical information. infform o ation. on This This constraint constraint is is currently currently being beeing and, addressed by the the database dattabbase MarinLit, Mari r nLit, with with the the first first group grooup with with geographical geograpphical data data da addressed availabble being being the the algae. algae. In this this presentation presentattion trends trends in in the the h discovery discovery of marine mari r ne algal algal available naatural product product are arre highlighted, highlighted, with with emphasis empphasis to the the identification ide d ntification of biodiscovery biodiscovvery natural hotspots, as as well well as as temporal tempporal and anndd spatial spaatial biodiscovery biodiscovery patterns. paatterns. Taxonomy, Taxonomy, chemistry chemistry hotspots, annd biogeography bioge o ograpphy are arre combined com mbbined for foor the the first first time time to to provide provide insights insights into into future future and bioprospe p cting efforts effort o s off unexplored unexplorred organisms orgaannisms and/or annd/or areas. areas. bioprospecting

52



Thursday 19th and Friday 20th September, 2013

Special Symposium for Young Investigators Special Symposium for Young Investigators AFTERNOON SESSION THURSDAY 19th Chair: William Fenical 17:00 - 17:40 PL 6 Brian Murphy 17:40 - 18:10 IL 10 Gordon Florence MORNING SESSION FRYDAY 20th Chair: William Fenical 09:00 - 09:30 IL 11 Antonio Hernández 09:30 - 09:50 OC 21 Robert Britton 09:50 - 10:10 OC 22 Robert Brkljaca 10:10 - 10:30 OC 23 Efstathia Ioannou 10:30 - 10:50 OC 24 Kine Østnes Hanssen 10:50 - 11:30 Coffee break 11:30 - 11:50 OC 25 Paula Jiménez 11:50 - 12:15 Closing speech: International Advisory Board Committee 12:15 - 12:30 Closing speech: Chair of the Congress 12:30 - 12:45 Closing speech: President of PharmaMar 12:45 - 13:15 Selection of the host for the 15th MaNaPro Symposium 13:15 - 14:30 Lunch

Illinois University, USA University of St. Andrews, UK

University of La Laguna, Spain Simon Fraser University, Canada Rmit University, Australia University of Athens, Greece University of Tromsø, Norway Labomar - Universidade Federal do Ceara, Brazil


53


PL 6

Afternoon Session

PLENARY LECTURE 6

LAKE MICHIGAN-DERIVED DIAZA-ANTHRACENE ANTIBIOTICS THAT TARGET Mycobacterium tuberculosis Chang Hwa Hwang2, Andrew Newsome1, Michael Mullowney1, Xiaomei Wei1, Larry Klein2, Urszula Tanouye 1, Minjee Kim2, Sanghyun Cho2, Scott Franzblau1,2, Brian T. Murphy1,2 1

Department of Medicinal Chemistry and Pharmacognosy and 2Institute for Tuberculosis Research, University of Illinois at Chicago, Chicago, IL. E-mail: [email protected]


In 2012 the World Health Organization estimated that there were 1.4 million deaths as a result of tuberculosis infection (TB), while one-third of the world’s population was infected with the etiologic agent. The most significant threats to the population are multidrug- and extensively drug-resistant strains of M. tuberculosis (MDR- and XDR-TB), which are resistant to first and second line drug regimens, respectively. Thus, novel drug scaffolds that can reduce TB spread and efficiently cure drug-resistant cases are in serious need. Our program focuses on harnessing the potential of freshwater-derived actinomycete bacteria as a source for potential TB leads. After screening our fraction library of actinomycete secondary metabolites against M. tuberculosis H37Rv using the microplate Alamar Blue Assay (MABA), we discovered that a Micromonospora sp. collected from Lake Michigan sediment (80 m) produced several submicromolar TB antibiotics. We cultivated this strain in 30 L of freshwater media, extracted the extracellular secondary metabolites, and after several chromatographic steps we isolated and identified a class of diaza-anthracene antibiotics that exhibit potent inhibitory activity toward mycobacteria. Some of these molecules displayed an in vitro activity profile similar or superior to clinically used TB agents with drug-like potency against a panel of drug-resistant TB strains, and less activity against non-mycobacteria, E. faecalis, S. pneumoniae, A. baumanii, P. aeruginosa, E. coli, and C. albicans. Synthetic efforts, in vitro ADMET and in vivo DMPK analyses, and preliminary steps toward molecular target identification will be discussed.

54



Afternoon Session

IL 10

INVITED IN VITED LECTURE LECTURE 1100

RESOLVING SYNTHESIS: LEIODOLIDE AND R ESOLVING STRUCTURE STRUCTURE THROUGH T THROUGH S YNTHESIS: LEI ODOLIDE A A ND PALMEROLIDE C PALMEROLIDE Gordon J. Florence Flloorence Gordon EaStCHEM Ea StCHEM School School of of Chemistry, Chemistry, University University ooff S Stt A Andrews, ndrews, United United Kingdom K in g d o m E-m mail: [email protected] [email protected] E-mail:


N ature provides p provi des an an armada arm r ada of structurally strructurally diverse diverse secondary seconddaarry metabolites metabbolites with with unique uniique Nature aand nnd often offten unexplored unexplored biological biological modes modes of action. action. Combined Com mbined with with their their molecular molecul u ar natural products aarchitectures rrchitectures na atural produc ts continue cont o inue to to provide provide the the inspiration insspiraation to to develop develop practical practical ssynthetic ynthetic routes routes and annd methods methods not n only only to to aid aid further fuurther biological b ological evaluation, bi evaluattion, on butt to to provide provi de confirmation conffirmation of their their complete complete structure. structure. This This talk talk will will discuss discuss our recent recent eendeavours ndeavoours towards towards r the syntheses syntheeses of two two marine marrine macrolides, macrolides, leiodolide leiodolide A and annd the palmerolide which properties pa lmeroolide C, w hich have haavve displayed displ p ayyed potent potent and and selective selective anticancer annticanncer prope rties in in pre limina n ry bi ological sstudies. tudi u es. preliminary biological


55


IL 11

Morning Session

INVITED LECTURE 11

STRUCTURAL STUDIES ON COMPLEX POLYKETIDES FROM DINOFLAGELLATES Antonio H. Daranas, Humberto J. Domínguez, José G. Napolitano, Francisco Cen Pacheco, Tamara S. Vilches, Guillermo Mendoza, Manuel Norte, José J. Fernández University Institute of Bioorganic Chemistry “Antonio González”, Department of Chemical Engineering and Pharmaceutical Technology, University of La Laguna. La Laguna, 38206, Canary Islands. Spain

Marine dinoflagellates produce many of the most active and complex secondary [1]

metabolites found in nature. Some of them have had an extraordinary impact upon different life science areas such as human health, pharmacology, analytical and natural product chemistry or even in the economics of fishery industry; promoting new developments in all these areas. In particular, dinoflagellates of the genus Prorocentrum, recognized as co-responsible of the DSP (Diarrhetic Shellfish Poisoning) syndrome, are well known to produce several unique bioactive secondary metabolites with a broad diversity of skeletons, including "linear" polycyclic [2-6]


compounds and macrolides. As a result of our continuous search for new bioactive products from artificial cultures of marine dinoflagellates, this communication deals with structural and biosynthetic features of complex polyketides isolated from cultures of Prorocentrum belizeanum

1. 2. 3. 4. 5. 6.

56

M. Murata, T. Yasumoto, Nat. Prod. Rep. 2000, 17, 293-314. T. Yasumoto, M. Murata, Y. Oshima, M. Sano, G. K. Matsumoto, J. Clardy, Tetrahedron 1985, 41, 10191025. K. Torigoe, M. Murata, T. Yasumoto, J. Am. Chem. Soc. 1988, 110, 7876-7877. C-K. Lu, H-N. Chou, C-K. Lee, T-H. Lee, Org. Lett. 2005, 7, 3893-3896. P. G. Cruz, A. H. Daranas, J. J. Fernandez, M. Norte, Org. Lett. 2007, 9, 3045-3048. P. G. Cruz, J. J. Fernandez, M. Norte, A. H. Daranas, Chem. Eur. J. 2008, 14, 6948-6956.



Morning Session

OC 21


CHLOROHYDRINS: VERSATILE BUILDING BLOCKS FOR THE SYNTHESIS OF MARINE NATURAL PRODUCTS Robert Britton,* Baldip Kang, Jason Draper, Stanley Chang, Michael Holmes, Hope Fan Department of Chemistry, Simon Fraser University, Burnaby, British Columbia, Canada, V5A 1S6 (ph: 001-778-782-4889; fax: 001-778-782-3765) E-mail: [email protected]


The formation of 1,2-anti-chlorohydrins via the addition of Gignard reagents to chloroaldehydes was first reported in 1959.1 That little has been done to exploit these intermediates since this time may largely be attributed to their perceived instability, preparation as diastereomeric mixtures (dr 4:1), and lack of practical methods to access these compounds in optically pure form. Recently, we have focused on identifying new roles for chlorohydrins (generated via catalytic asymmetric chlorination2) in natural product synthesis, and have found that they engage in highly diastereoselective reactions with a variety of organolithium reagents.3 These findings now enable rapid access to variously functionalized and enantiomerically enriched heterocycles, including: epoxides, tetrahydrofurans, pyrrolidines, pipiridines, and spiroacetals. A survey of the methods developed to access these heterocycles from chlorohydrins, as well recent examples of their application to the rapid synthesis of marine natural products will be presented.

References 1. 2.

3.

Cornforth, J. W.; Cornforth, R. H.; Mathew, K. K. J. Chem. Soc. 1959, 112. For the asymmetric -chlorination of aldehydes see: (a) Brochu, M. P.; Brown, S. P.; MacMillan, D. W. C. J. Am. Chem. Soc. 2004, 126, 4108. (b) Halland, N.; Braunton, A.; Bachmann, S.; Marigo, M.; Jørgensen, K. A. J. Am. Chem. Soc. 2004, 126, 4790. (c) Amatore, M.; Beeson, T. D.; Brown, S. P.; MacMillan, D. W. C. Angew. Chem. Int. Ed. 2009, 48, 5121. (a) Kang, B.; Britton, R. Org. Lett. 2007, 9, 5083. (b) Kang, B.; Mowat, J.; Pinter, T.; Britton, R. Org. Lett. 2009, 11, 1717. (c) Kang, B.; Chang, S.; Decker, S.; Britton, R. Org. Lett. 2010, 12, 1716. Draper, J.; Britton, R. Org. Lett. 2010, 12, 4034; (d) Halperin, S.; Kang, B.; Britton, R. Synthesis 2011, 12, 1946; (e) Chang, S.; Britton, R. Org. Lett. 2012, 14, 5844; (f) Britton, R.; Kang, B. Nat. Prod. Rep. 2013, 30, 227; (g) Halperin, S.; Britton, R. Org. Biomol. Chem. 2013, 11, 1702; (h) Dhand, V.; Draper, J. A.; Moore, J.; Britton, R. Org. Lett. 2013, 15, 1914.


57


OC 22

Morning Session


PHYTOCHEMICAL PROFILING AND BIOLOGICAL ACTIVITY STUDIES OF THE SOUTHERN AUSTRALIAN MARINE ALGA, Sargassum paradoxum Robert Brklja a and Sylvia Urban Marine and Terrestrial Natural Product (MATNAP) Research Group, School of Applied Sciences, Health Innovations Research Institute (HIRi) RMIT University, GPO Box 2476V Melbourne, Victoria 3001, Australia. E-mail: [email protected]

In our continuing effort to discover new bioactive natural products from southern Australian marine algae, a specimen of the brown alga, Sargassum paradoxum, which displayed anti-microbial activity, was investigated. Marine algae of the genus Sargassum are known for producing chromenes, chromanols, quinones and hydroquinones.1,2


A chemical profiling study employing on-flow and stop-flow HPLC-NMR together with HPLC-MS methodologies was implemented to rapidly identify multiple new and previously described sesquiterpenes.3 Importantly HPLC-NMR was able to identify a new unstable sesquiterpene that could not be isolated by off-line methodologies. Subsequent off-line bioassay-guided fractionation resulted in the isolation of seven new and five previously reported sesquiterpenes. All isolated sesquiterpenes showed various degrees of activity. The absolute configuration of three of these compounds was proposed on the basis of Circular Dichroism (CD) Spectroscopy. This study was able to demonstrate the advantage of using hyphenated spectroscopic techniques like HPLC-NMR and HPLC-MS for the rapid chemical profiling/dereplication of crude extracts. References 1.

58

MarinLit Database (2012), University of Canterbury, Christchurch, New Zealand.

2.

Reddy P. and Urban S. Phytochemistry (2009), 70, 250-255.

3.

Brklja a R. and Urban S. Journal of Natural Products (Manuscript in preparation).



Morning Session

OC 23


NEW POLYKETIDES FROM A MARINE-DERIVED ACTINOBACTERIUM Eniko Rab1,2, Dimitris Kekos2, Vassilios Roussis1, Efstathia Ioannou1 1

University of Athens, School of Pharmacy, Department of Pharmacognosy and Chemistry of Natural Products, Panepistimiopolis Zografou, Athens 15771, Greece. 2National Technical University of Athens, School of Chemical Engineering, Biotechnology Laboratory, 9 Iroon Polytechniou Str., Zografou Campus, Athens 15780, Greece E-mail: [email protected]


In the ongoing search for more effective anticancer and antibiotic agents, microorganisms have emerged as a goldmine yielding secondary metabolites with potent biological activities. Even though terrestrial habitats have been the major source of such microorganisms, the world’s oceans have been largely understudied in this regard. Nonetheless, over the last fifteen years marine-derived actinobacteria have been proven to produce a wide variety of structurally diverse bioactive natural products. In order to obtain new strains likely to produce novel metabolites, examination of samples from different habitats and extreme environments is necessary. The East Mediterranean basin is a geomorphologically and biologically unique marine ecosystem that has not been investigated so far for its microbiota, especially from the chemical point of view. n this context, we turned our attention to an actinobacterial strain which exhibited an interesting profile during a preliminary LC-MS screening. The strain was cultured in flasks containing a seawater-based medium for 7 days. Subsequently, Amberlite XAD-7 resin was added to each flask and the slurry was shaken for 6 h. The resin was filtered through cheesecloth, washed with deionized water and extracted with acetone. Evaporation of the solvent afforded the crude extract which was subjected to a series of chromatographic separations that has led so far to the isolation of twelve secondary metabolites. The structures of the isolated compounds were determined on the basis of extensive analyses of their spectroscopic data (NMR, MS, IR, UV). Among them, two are new natural products bearing the rare wailupemycin skeleton. Acknowledgements: This work is supported by the project “BIOMARACT”, which is implemented under the “ARISTEIA” Action of the Operatıonal Programme “EDUCATION AND LIFELONG LEARNING” and is co-funded by the European Social Fund (ESF) and National Resources.


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OC 24

Morning Session


INTRODUCING ATOMIC FORCE MICROSCOPY IN STRUCTURE ELUCIDATION - THE DISCOVERY OF BREITFUSSIN A AND B, HIGHLY MODIFIED HALOGENETED DIPEPTIDES FROM THE ARCTIC HYDROZOAN Thularia breitfussi Kine Østnes Hanssen1, Bruno Schuler2, Antony J. Williams3, Taye B. Demissie4, EspenHansen1, Jeanette H. Andersen5, Johan Svenson5, Leo Gross2, Marcel Jaspars5, Johan Isaksson5 1

Mabcent-Sfi, University of Tromsø, N-9037, Tromsø, Noruega, 2IBM Research, Rüschlikon, Zurich, 8803, Switzerland, 3Royal Society of Chemistry, 904 Tamara Circle, Wake Forest, 27587, USA, 4Centre For Theoretical And Computational Chemistry, University of Tromsø, Tromsø, N-9037, Norway, 5Marine Biodiscovery Centre, Department of Chemistry, University of Aberdeen, Old Aberdeen, AB24 3UE, UK. E-mail: [email protected]


Atomic force microscopy (AFM) with atomic resolution has for the first time been used to aid in the elucidation of unknown natural products (Angew Chem Int Ed Eng 2012, 51, (49), 12238-41). AFM shows great potential for the structural characterization of planar, proton-poor compounds, as these compounds are difficult to elucidate with spectroscopic methods and therefore prone to corrections. Given the limited quantity isolated, x-ray crystallography was not possible and structure determination using spectroscopic data was unable to propose a single unequivocal structure consistent with all the data. We show here the combined use of spectroscopic methods, AFM, computer assisted structure elucidation (CASE) and electronic structure calculations (DFT) to solve the structures of breitfussins A and B, isolated from the Arctic hydrozoan Thuiaria breitfussi (Family Sertulariidae), which could not be solved using either method alone. This study presents the first compounds from the hydrozoan Thuiaria breitfussi, revealing an unusual structural framework containing a combination of indole-oxazole-pyrrole. Remarkably, AFM could be used to determine all the connection positions of the cyclic systems as well as those of the substituent groups: MeO, Br, and I information that is difficult to obtain with other techniques.

60



Morning Session

OC 25


CYTOKINESIS BLOCKAGE INDUCED BY A DITHIOLPYRROLONE ISOLATED FROM A Streptomyces sp. STRAIN RECOVERED FROM THE BRAZILIAN ENDEMIC TUNICATE Eudistoma vannamei Paula C. Jimenez1,2*, Paula A. Abreu1,2, Thiciana S. Sousa3, Danilo D. Rocha1, Diego V. Wilke1, Edilberto R. Silveira3, Otília D. L. Pessoa3, James J. La Clair4, Letícia V. Costa-Lotufo1,2 1

Departamento de Fisiologia e Farmacologia – Universidade Federal do Ceará (UFC), Fortaleza, CE, Brazil. 2Instituto de Ciências do Mar - UFC, Fortaleza, CE, Brazil; 3Departamento de Química Orgânica e Inorgânica – UFC, Fortaleza, CE, Brazil. 4Xenobe Research Institute, San Diego, California, USA E-mail: [email protected]



Marine microorganisms are known to produce a variety of interesting cytotoxic and antitumor compounds. As an effort to study Brazilian species, we examined the microorganisms recovered from the endemic tunicate Eudistoma vannamei as a potential source for new anticancer leads. One strain, identified as Streptomyces sp., was found to produce the dithiolpyrrolone N-(4,5-dihydro-5-oxo-1,2-dithiolo[4,3b]pyrrol-6-yl)-N-methyl-formamide. This compound presented IC50 values ranging from 1.7-6.4μM across a panel of tumor cell lines, with the more potent activity arising in PC-3/M, a resistant human metastatic prostate carcinoma cell line. We then turned our attention to interrogate its putative biological action. Beginning with flow cytometry, we found that the dithiolpyrrolone induced cell cycle arrest during mitosis after 24h or 48h exposure, an observation that was further confirmed by the increase of the mitotic index of treated PC-3/M cells with concentrations varying from 0.5-5μM. Confocal microscopy imaging confirmed the blockage during mitosis, evidenced by cells displaying incomplete cell division. The presence of hypertetraploid cells, suggesting unfinished transitioning through cytokinesis, further illustrated this phenotype. Additional analyses via western blot indicated that dithiolpyrrolone-treated cells showed a reduced expression of PRC-1, PLK-1 and RhoA, proteins that are necessary for the cleavage furrow assembly and exit from cytokinesis. The evidences herein suggest that the dithiolpyrrolone impaired the progression through cytokinesis by modulating the function of kinetic proteins, such as PLK-1, that are required to be aligned in the equator of the cell by the end of anaphase in order to complete successful passage through cytokinesis.

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Cubierta 4:Abstracts 01/08/13 20:01 Página 1





September 15th - 20th, 2013 La Toja Island, Galicia Organized by:

Spain

Programme and Abstracts

on Marine Natural Products - MANAPRO 2013

on Marine Natural Products - MANAPRO 2013

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