Adrianna Ianora Stazione Zoologica Adrianna Ianora Stazione Zoologica ““Anton Anton DohrnDohrn”” NapoliNapoli

The Potential of Marine Microorganisms and the Use of Chemical Ecology-Derived Methods for Drug Discovery

Advantages of marine biodiversity

Current Commercial Marine Drugs

Clinical trial Name Source Target Developed byIn clinical use ectenaiscidin 743 (Yondelis) tunicate tubulin PharmaMar, RinehartPhase III E7389 (halichondrin B inspired)* synthetic tubulin EisaiPhase II dehydrodidemnin B (Aplidine) tunicate ornithine decarboxylase PharmaMar, RinehartPhase II soblidotin (aka TZT1027, dola‐10 insp.) synthetic tubulin Teikoku, PettitPhase II synthadotin (akaILX651, dola‐15 insp.) synthetic tubulin ILEXPhase II bryostatin 1 bryozoan PKC GPC Biotech, PettitPhase II squalamine shatk angiogenesis ZasloffPhase II kahalalide F mollusk multiple PharmaMar, RinehartPhase I PM02734 (kahalalide insp.) synthetic solid tumor PharmaMarPhase I Zalypsis (jorumycin insp.)* synthetic DNA PharmaMarPhase I E7974 (hemiasterlin insp.)* synthetic tubulin EisaiPhase I taltobulin (aka HTI286, hemiasterlin insp.)* synthetic tubulin Wyeth, AndersenPhase I salinosporamide A (aka NPI0052) bacteria proteasome Nereus, FenicalPhase I spisulosine (aka ES285) clam Rho PharmaMarPhase I KRN‐7000 (agelasphin insp.)* synthetic NKT Koezuka‐KirinPhase I NPI2358 (halimide insp.) synthetic tubulin Nereus, FenicalPhase I LBH589 (psammaplin insp.)* synthetic HDAC NovartisDiscontinuedPhase II <2004 dolastatin 10 sea  hare tubulin PettitPhase II<1999 didemnin B tunicate antineoplastic RinehartPhase II <2004 cemadotin (dola‐15 insp.) synthetic tubulin BASF, PettitPhase II<2002 cryptophycin 52 (arenastatin)* synthetic tubulin Lilly, ValeriotePhase I<2004 discodermolide* sponge tubulin Novartis, HBOIPhase I<2002 LAF389 (bengamide insp.) synthetic MetAP Novartis, CrewsPhase I<2006 LAQ824 (psammaplin insp.) synthetic HDAC Novartis, CrewsPhase I<2000 girolline (aka girodazole)* sponge protein synthesis Potier* substances from marine sponges

Marine Drugs other than Antitumors

‐as nutritional supplements including color additives and antioxidants‐vitamins, oils, and cofactors which enhance general well‐being ‐The carotenoid market alone was projected to reach77 million Euro by 2010 

Dunaliella salina

Sources of Marine Natural Products

Cultured strain of Lyngbya majuscula L. majuscula at high magnification

Colonies of a marine cyanobacterium  from reefs in Papua New Guinea

Collection of filaments of a marine cyanobacterium into plastic bags

Gerwick et al. 2008

Nearly 800 compounds Have been reportedfrom marine cyanobacteria

Gerwick et al. 2008

In pre‐clinical trials

MARINE PHYTOPLANKTON AS POTENTIAL NEW DRUGS

Unicellular algae that are at the base of the marine food chain 

There are more than 200 genera of living diatoms, and it is estimated that there are approximately 100,000  speciesDiatoms can be found in the oceans and in freshwater Most live pelagically in open water, although some live as surface films at the water‐sediment interface (benthic), or even under damp atmospheric conditions They are especially important in oceans, where they are estimated to contribute up to 45% of the total oceanic primary production.

Miralto et al. Nature 1999Miralto et al. Nature 1999

Oxylipin and aldehyde methabolism in diatoms

6-LOXHTrA

CO2H

H OOHCO2H

H OHCO2H

CO2HH OOH

CO2HO

CO2HH OH

6-LOX

CO2H

H OOHCO2H

H OHCO2H

CO2HH OOH

CO2HO

CO2HH OH

9-LOX

9-LOX

HTA

O

O

CO2H

CHO

CO2H

EPA11R-LOX

CO2H

HOO

CO2H

HO

12-LOX

CO2HOOH

CO2H

CO2HOH

O

OH

decatrienal

octatrienal

octadienal

C16-PUFAs

C20-PUFAs

Fontana et al. 2007Fontana et al. ChemBioChem 2007

O

OOH

OH

LOX

OH

O

CO2H

ROS

Lipolysis

Fatty acid hydroxide

Epoxy alcohol fatty acid

Polyunsaturated aldehyde

Fatty acid hydroperoxide

Fatty acid

Fontana et al. 2007

COOH

Plants

18:3ω3/18:2ω6

Blee et al. 2002

So what is the function of these metabolites?

Chemical Defense

- Noxious compounds act as feeding deterrents.

-Their main purpose would not be to intoxicate but to discourage tasting.

- Deterrence could occur after ingestion of prey cells and the predator response could be regurgitation or discouragement of further feeding

What happens if the predator feeds on toxic cells?

physical incapacitation (paralysis, altered swimming behaviour etc..) or mortality of the predator

reduction in fecundity or depressed viability of gametes

In either case, blooms grow and persist when grazing pressure would otherwise have caused them to crash

Terrestrial environmentTerrestrial environment1) Coumarins (parsley)2) Isoflavonoids (clover)3) Azadirachtin ( neem tree)4) Phytojuvenoids (sweet basil)5) Sesamin (sesame oil)6) Nicotine (tabacco)

TeratogenicTeratogenic compoundscompounds

Aquatic environmentsAquatic environments1) Aldehydes (diatoms)

Birth control effectBirth control effect

Nicotine

How do we use this chemical ecology-derived information for drug discovery?

Fontana et al. 2007

Hatching failure in the copepod Temora stylifera due to somedinoflagellate diets (Prorocentrum micans, Gymnodinium sanguinium and Gonyaulax polyedra)

Eggs stained with Hoechst 33342

Ianora et al. Limnol. Oceanogr. 1999

Harmful Algal Blooms

Saxitoxin

Alexandrium tamarense

Types of chemical ecology‐ derived bioassays :

• chemicals used to capture prey or to deter grazers• Allelopathy chemicals produced by microalgae to deter 

growth of competitors• Pathogen‐host interactions• Chemicals that alter reproduction and development of 

predators

We need to better foster and support the development of the new emerging field of marine chemical ecology

Proven antifeedants have been used to derive anti‐cancer, anti‐viral and anti‐agingproducts; toxins have been used to develop pain‐killers; and functional products have been obtained from the materials used by sessile organisms to colonize free surfaces. 

Knowledge of eco‐physiological interactions may serve as a platform to facilitate the search for new biotechnological candidates, as well as to optimize culture conditions and achieve production of biomass for industrial applications.

More research needs to be focused on the identification of metabolites that are active in the ecological context. Once in hand the few available compounds have been drivers of intensive research efforts that have resulted in new concepts and advancements in the field. 

•If we can understand the natural function of these compounds, we will be able to manage and protect them better, and find new biotechnological applications for these potentially important natural products in the future

Massimo PernaTechnician

Dott. Antonio MiraltoResp. Lab. Ecofisiologia

D.ssa Adrianna IanoraSenior Scientist

D.ssa Isabella ButtinoAssociate Scientist

D.Ssa Raffaella CasottiAssociate Scientist

Andrea Gerecht PhD student

D.SsaGiovanna RomanoAssociate Scientist

Mario Di PintoTechnician

Dott. Francesco EspositoTechnologist Vittoria Roncalli

MSc student

D.ssa Ylenia CarotenutoAssociate Scientist

Flora PalumboTechnician

Francois RibaletPhD student

Cecilia Balestra Phd student

CNR Istituto di Chimica Biomoleculare

CNR-ICBAngelo FontanaAdele “Lella” CutignanoGiuliana d’IppolitoNadia LamariAntonio MaielloGuido Cimino

CNR-ISAGianluca Picariello

CNR-IBPNando Febbraio