Aveiro, November 25-27, 2012

Natural Products and related Redox Catalysts: Basic Research and Applications in Medicine and Agriculture

Natural Products and related Redox Catalysts: Basic

Research and Applications in Medicine and Agriculture

BOOK OF ABSTRACTS

Front Cover Design: Diana Pinto Photograph: “Canal S. Roque” by José M. G. Pereira Structure: Quercetin

Natural Products and related Redox Catalysts: Basic Research and Applications in Medicine and Agriculture

Scientific Committee

Artur M. S. Silva, Department of Chemistry and QOPNA, University of Aveiro Claus Jacob, School of Pharmacy, Universität des Saarlandes (USAAR) Diana C. G. A. Pinto, Department of Chemistry and QOPNA, University of Aveiro Gilbert Kirsch, Laboratoire d’Ingénierie Moléculaire et Biochimie Pharmacologique, Université de Metz (UPV-Metz)

Organizing Committee

Artur M. S. Silva (Chairman – Dep. Chem., Univ. Aveiro) Augusto A. C. Tomé (Dep. Chem., Univ. Aveiro) Diana C. G. A. Pinto (Dep. Chem., Univ. Aveiro) Mª Graça P. M. S. Neves (Dep. Chem., Univ. Aveiro) Graça M. S. Rocha (Dep. Chem., Univ. Aveiro) José A. S. Cavaleiro, (Dep. Chem., Univ. Aveiro) Mª do Amparo F. Faustino (Dep. Chem., Univ. Aveiro) Mário M. Q. Simões (Dep. Chem., Univ. Aveiro) Aurora Fernandes (Dep. Chem., Univ. Aveiro)

Sponsors

Contents

Welcome .............................................................................. 3

A warm Word of Welcome from the RedCat Coordinator ... 5

Scientific Program ............................................................... 7

Plenary Lectures ................................................................ 11

Invited Lectures ................................................................. 21

Oral and Poster Communications ..................................... 27

Authors Index ..................................................................... 61

Participants List ................................................................ 67

3

Welcome

RedCat is an Initial Training Network for early stage and experienced researchers

funded under the Framework 7 ‘People’ programme of the European Union. RedCat

started on December 2008 and will be ended on November 2012.

RedCat provides research and training opportunities for 10 early stage and 4

experienced researchers across Europe, with 10 partner institutions and 8 associated

partners in 5 European countries (Germany, France, Luxembourg, Portugal, UK). RedCat

conducts its own research, which addresses highly significant, up-to-date research

questions in the area of natural product research, intracellular redox processes, drug

development and ‘green’ agriculture. Individual projects run across scientific disciplines.

They embrace chemistry, biochemistry, biology, pharmacy, medicine and agricultural

research.

RedCat researchers receive extensive training in research methods and research

related subjects, including bioethics, intellectual property and communication. Training is

provided by experts in the field, at excellent institutions with the most modern equipment

and techniques. The aims of the training provided Red Cat are summarized in its motto: Fit

for Europe, fit for the future .

The RedCat ITN project considered two main scientific meetings: 1) The “Redox

regulation - natural compounds as regulators of inflammation signalling” meeting occurred

last January (January 25-27, 2012), in Luxembourg (Marc Diederich as chairman) and

having 130 participants; 2) The present meeting is the second and last one, and will be held

at the University of Aveiro (November 25-27, 2012; at the Rectory building) as a satellite

of the 3rd Portuguese Meeting on Medicinal Chemistry.

I promise you that I and the members of the Organizing Committee will do

everything to make your visit comfortable and gratifying and we welcoming you to the

Natural Products and related Redox Catalysts: Basic Research and Applications in

Medicine and Agriculture at Aveiro during November 25-27, 2012.

The Chairman Artur Silva

4

5

A warm Word of Welcome from the RedCat Coordinator

Dear friends,

as you all know, this meeting of our RedCat network will be the last of its kind. It is

therefore my pleasure to welcome you to this meeting - which in many ways represents the

culmination of our RedCat activities, and also to the wonderful city of Aveiro. At the same

time, it is also the time for me to say goodbye to all of you after four years of close

interactions and some really exciting research we have conducted together.

When the RedCat idea was developed a couple of years ago, we decided to use the

motto “Fit for Europe, fit for the Future” as a kind of overarching statement. In hindsight,

this choice was probably better than we anticipated at the time. Many of you have travelled

throughout Europe (some by plane or car, some by donkey), visited and even stayed in

countries which you knew little of before. The mobility and personal experiences achieved

as part of RedCat have been truly breathtaking, some of you have even been robbed at pen-

point in eternal cities or turned into cunning linguists as part of your training. Ultimately,

some of you have really become fit for Europe and subsequently have found new positions,

employment and careers in countries across the EU. Despite the fact that Europe has

become a bit unpopular lately, the RedCats have shown that the European spirit is alive and

can be turned into something wonderful.

Whilst this may be seen as the key success of RedCat, we can also be proud of our

training, which has not just covered purely scientific issues, but also other subjects

including the social sciences (remember the debate in the English pub or the artist with her

little fishies), communication, IPR and commercialization - and how to “go to Macedonia”

or to wear a “masque à gaz avec cartouche”, to name just a few. I still believe that this kind

of training has been important as it has really given some of you a “cutting edge” over

some traditionally trained “lab rats”.

And then, of course, we have conducted some “rather stimulating research”, and

published a “honey-waggon full of 1A-prime quality s**t” as David and I would put it,

albeit respectively. In fact, we have outperformed our most ambitious goals, in number as

well as quality of original papers and reviews, and we are even working on a joint RedCat

book right now. Some of you are just completing their PhDs, and again, the efforts you

have put into your research, and the amazing success stories we will hear about at this

meeting, go far beyond of everyone’s expectations.

6

I know that you also expect from me a word on “unisex”. Well, this is a great concept

and I am particularly proud to tell you that despite the involvement of two RedCats with

rather phallistic surnames, our network has provided training for a number of highly

successful and promising female young investigators, who surely will have a great career in

front of them. Our network has also trained a couple of non-EU researchers, who have

enjoyed their time in Europe, or, in some instances, also their time of being bullied in some

European country embassies applying for visa, and hopefully will stay in Europe and

contribute to European science in the future. For these students, in particular, the RedCat

actions have been extremely valuable as a guide to Europe, its culture, habits and pork

sausages.

At this final meeting, you will also meet our external advisors, some of whom have

long been waiting for the chance to meet all of you. The fact that we could not feed them

(in) at an earlier stage was one of the few koeck-ups which occurred during the planning of

the network and I take full responsibility for this. We simply did not anticipate and hence

budget the costs correctly for the earlier meetings. Indeed, not everything has worked

according to plan, but most things have and I hope that you have enjoyed the RedCat as

much as I have.

One item that was scheduled from the very beginning, however, is this meeting. We

were sure that Aveiro would be the most appropriate place for our farewell meeting, as it is

one of the most beautiful cities we can offer and just right for the kind of firework of

science we surely can expect during the next couple of days. So let’s enjoy this meeting,

take stock of the past but, above all, also look towards the future which, for sure, will see

some small baby cats continuing in the footsteps of the old RedCat.

Welcome to the meeting and all the best for the future,

The Coordinator of the RedCat Project

Claus Jacob

Scientific Program

SCIENTIFIC PROGRAM

8

Scientific Program

Sunday, November 25th

14:00-16:00 Registration

16:00-16:30 Opening Ceremony

Chairperson: Artur Silva

16:30-17:20 PL1 – Eric Block

Department of Chemistry, University of Albany, Albany, New York, USA Synthesis and characterization of polysulfanes with twenty or more sulfur atoms through use of liquid sulfur as a reagent: applications to the chemistry of garlic

17:20-17:50 IL1 – Alan Slusarenko

Department of Plant Physiology, RWTH Aachen University, Aachen, Germany Control of plant diseases with natural products: allicin from garlic as an example

17:50-18:15 OC1 – Awais Anwar

ECOspray Limited, Grange Farm Hilborough, Norfolk, United Kingdom Garlic in agriculture - developments and challenges

18:15-18:40 OC2 – Zhanjie Xu

Ursapharm Arzneimittel GmbH, Saarbrücken, Germany Synthesis and biological activity of 1H-naphtho[1,2-c]chromene-1,4,5-trione and benzo[i]phenanthridine-1,4,5(6H)-trione derivatives

19:00- ... Welcome reception

Monday, November 26th

Chairperson: Artur Silva

09:00-09:50 PL2 – Péter Nagy

Department of Molecular Immunology and Toxicology, The National Institute of Oncology, Budapest, Hungary Kinetics and mechanisms of thiol oxidation in biological systems

09:50-10:40 PL3 – Clementina Santos

School of Agriculture, Polytechnic Institute of Bragança, Bragança, Portugal; Department of Chemistry & QOPNA, University of Aveiro, Aveiro, Portugal Polyhydroxylated 2-styrylchromones and -xanthones as potent antioxidant agents

10:40-11:00 Coffee break

11:00-11:25 OC3 – Oualid Talhi

Department of Chemistry & QOPNA, University of Aveiro, Aveiro, Portugal Michael addition on chromone-3-carboxylic acid: a one-pot tandem reaction towards novel polysubstituted -chromones, -chromanones, and –flavones

11:25-11:50 OC4 – Lidia Brodziak-Jarosz

German Cancer Research Center (DKFZ), Heidelberg, Germany Xanthohumol from hops activates the cytoprotective Keap1-Nrf2 pathway through its enone site

9

11:50-12:15 OC5 – Clemens Zwergel

LIMBP, Institut Jean Barriol, Université de Lorraine, Metz, France Synthesis and biological evaluation of aurones and chalcones from coumarins and chromones

12:15-14:00 Lunch

Chairperson: Augusto Tomé

14:00-14:50 PL4 – M. Graça Neves

Department of Chemistry & QOPNA, University of Aveiro, Aveiro, Portugal Development of new tetrapyrrolic macrocycles with potential application in medicine

14:50-15:15 OC6 – Ifeanyi Nwachukwu

RWTH Aachen University, Department of Plant Physiology (BioIII), Aachen, Germany Oxidative stress in bacteria: differential sensitivities to selected oxidizing agents

15:15-15:40 OC7 – Maria Letizia Lo Faro

University of Exeter Medical School, Exeter, United Kingdom Nitric oxide and hydrogen sulfide cross-talk: regulation of bioavailability and cell signalling

15:40-17:00 Poster session and coffee break

Chairperson: Paul Winyard

17:00-17:30 IL2 – Marc Diederich

LBMCC, Hôpital Kirchberg, Luxembourg and College of Pharmacy, Seoul National University, Seoul, Korea Natural compounds as inhibitors of the 10 hallmarks of cancer

17:30-18:00 IL3 – Claus Jacob

Bioorganic Chemistry, School of Pharmacy, Saarland University, Saarbruecken, Germany Redox signalling via the cellular thiolstat - The special relationship in Group 16

18:00-18:25 OC8 – Nathaniel Saidu

Medical Biochemistry and Molecular Biology, University of the Saarland, Homburg, Germany Effect of diallyl tetrasulfide on HCT116 colorectal cancer cells

18:30- … Supervisory board meeting

20:00- … Congress dinner

Tuesday, November 27th

Chairperson: Gilbert Kirsch

09:00-09:50 PL5 – Anake Kijjoa

Instituto de Ciências Biomédicas de Abel Salazar and Centro Interdisciplinar de Investigação Marinha e Ambiental (CIIMAR), Universidade do Porto, Portugal Drugs, nutraceuticals and cosmeticals from marine sources

10

09:50-10:40 PL6 – Antonello Mai

Department of Drug Chemistry and Technologies, Sapienza University of Rome, Rome, Italy Design, synthesis, and biological evaluation of SIRT modulators

10:40-11:00 Coffee break

11:00-11:25 OC9 – Miriam Arbach

School of Pharmacy, University of East Anglia, Norwich, United Kingdom Diallyl polysulfides from garlic - mode of action and applications in agriculture

11:25-11:50 OC10 – Wioleta Marut

Université Paris Descartes, Faculté de Médecine, Hôpital Cochin AP-HP, Paris, France The natural organosulfur dipropyltetrasulfide prevents systemic sclerosis in the mice

11:50-12:15 OC11 – François Gaascht

LBMCC, Hopital Kirchberg, Luxembourg New natural compounds of Dionaea muscipula as anticancer agents

12:15-14:00 Lunch

Chairperson: Diana Pinto

14:00-14:25 OC12 – Diana Resende

Department of Chemistry & QOPNA, University of Aveiro, Portugal Exploring the versatility of (E,E)-cinnamylideneacetophenones as templates for the synthesis of potentially bioactive compounds

14:25-14:50 OC13 – Sergio Valente

Dipartimento di Chimica e Tecnologie del Farmaco, Sapienza Università di Roma, Roma, Italy 4-Vinyl-2H-1-benzopyran-2-one reactivity as [4+2] thermal Diels-Alder cycloaddition: access to novel coumarin-based polycycles with Cdc25 phosphatases inhibiting activity

14:50-15:20 IL4 – Chris Hamilton

School of Pharmacy, University of East Anglia, Norwich Research Park, Norwich, United Kingdom Bacillithiol:- The emerging redox properties and functions of the glutathione surrogate in many Gram positive bacteria

15:20-16:30 Poster session and coffee break

Chairperson: Claus Jacob

16:30-17:20 PL7 – Richard Glass

Department of Chemistry, University of Arizona, Tucson, Arizona, USA Redox chemistry of sulfur compounds and its biological relevance

17:20-17:30 Closing ceremony

Plenary Lectures

PLENARY LECTURES

PL1

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Synthesis and characterization of polysulfanes with twenty or more sulfur atoms through use of liquid sulfur as a reagent: applications to the

chemistry of garlic

Eric Block,a Murree Groom,b Robert Sheridan,c Kai Wang,a Shaozhong Zhanga

aDepartment of Chemistry, University at Albany, State University of New York, Albany, New York, 12222,

United States; bECOspray Limited, Grange Farm, Hilborough, Norfolk, IP26 5BT, United Kingdom; cNYS

Department of Agriculture and Markets, Food Laboratory Division, Albany, New York, 12235, United States

Diallyl disulfide reacts within minutes with liquid sulfur at 120 °C giving a family of diallyl polysulfanes, All2Sn (n = 3–22), characterized by ultra-performance liquid chroma-tography-(Ag+)-coordination ion spray-mass spectrometry (UPLC-(Ag+)CIS-MS).[1] Similarly, garlic oil (GO), bis-(2-methyl-2-propenyl), bis-(2-chloro-2-propenyl), bis-(3-methyl-2-butenyl), and bis-(2-cyclohexen-1-yl) disulfides all give families of polysulfanes with up to 22 sequential sulfur atoms. New members of families of silver chelators with up to 10 sulfur atoms were found in GO using UPLC-(Ag+)CIS-MS.[2]

Figure. HPLC trace of reaction product of liquid S8 and diallyl polysulfane. The vertical axis corresponds to the approximate relative abundance of each peak based on UV absorbance. The number of sulfur atoms is listed following “DAS” in each case. Acknowledgments: This work was supported by the U.S. National Science Foundation (CHE-0342660, CHE 0450505, and CHE-0744578). Initial phases of this research were performed by E.B. and M.G. at the UK laboratories of ECOspray Ltd; E.B. thanks ECOspray for hosting his visits in the UK. Queries regarding GO-based pesticides should be directed to M.G.: groom@ecospray.com. References [1] Wang, K.; Groom, M.; Sheridan, R.; Zhang, S.; Block, E. J. Sulfur Chem., 2012, earlypub.

http://dx.doi.org/10.1080/17415993.2012.721368. [2] Block, E. Garlic and Other Alliums: The Lore and the Science, Royal Society of Chemistry, Cambridge,

454 pp.: 2010.

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Kinetics and mechanisms of thiol oxidation in biological systems

Péter Nagy

National Institute of Oncology, Hungary, Department of Molecular Immunology and Toxicology.

Cysteine residues have a wide range of physiological functions, which are mostly governed via the redox chemistry of their thiol functional group. Due to the nucleophilic nature of sulfur, thiol compounds are particularly susceptible to oxidation, which makes cysteines primary targets of endogenous oxidants. Thiols are efficient scavengers of radical species and they also engage in two electron oxidation reactions in vivo. Therefore, the thiol-disulfide pool is thought to regulate redox homeostasis, a thermodynamic parameter that determines which reactions can occur in cellular compartments. However, it is the kinetics of the reactions that determines cell dynamics, because the partitioning of the possible reactions depends primarily on kinetic parameters.

During cysteine oxidation, due to the promiscuous chemical properties of the thiol sulphur, a plethora of different products and intermediate species can form. In addition, the lack of distinct spectroscopic signatures of the various thiol derivatives, as well as the large scale of chemical factors that influence the reaction rates poses challenges for kinetic investigations.

This presentation is aimed to give insight into the advances and challanges that are associated with mechanistic studies of the redox reactions of thiols with biological oxidants. Among others I will discuss: i) the thiol pKa (which is an important parameter that largely influences nucleophilicity and reactivity) from a chemical perspective ii) the importance of knowing the nature of the reaction partner, the oxidant and iii) that primary reactions generate reactive intermediates, which initiate a cascade of redox events.

Kinetic studies of proteins are more challenging than small molecules, and therefore they often require a different approach. As an example, our mechanistic study on the peroxidise activities of the ubiquitous antioxidant thiol-proteins, peroxiredoxins 2 and 3, will be demonstrated. And finally the power of kinetic simulations in predicting the partitioning of oxidants between cellular thiols will be mentioned. Acknowledgments: The financial support of FP7-PEOPLE-2010-RG (Marie Curie International Reintegration Grant; grant No.: PIRG08-GA-2010-277006) is greatly acknowledged. References The subject of this presentation is reviewed in:

[1] Péter Nagy. Kinetics and Mechanisms of Thiol-Disulfide Exchange Covering Direct Substitution and Thiol Oxidation-Mediated Pathways Antioxid. Redox Signal. Thiol-Disulfide Exchange Forum Issue (2012) in press.

[2] Péter Nagy and Christine Winterbourn. Redox Chemistry of Biological Thiols Adv. Mol. Toxicol. Fishbein, J.C., Ed. Elsevier: Amsterdam, The Netherlands, (2010), Vol. 4, pp. 183-222.

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Polyhydroxylated 2-styrylchromones and -xanthones as potent antioxidant agents

Clementina M. M. Santos,a,b Artur M. S. Silva,b Diana C. G. A. Pinto,b José A. S.

Cavaleirob

aDepartment of Vegetal and Production Technology, School of Agriculture, Polytechnic Institute of Bragança, 5301-855 Bragança, Portugal; bDepartment of Chemistry & QOPNA, University of Aveiro, 3810-

193 Aveiro, Portugal

Chromones and xanthones are two classes of oxygenated heterocyclic compounds widely distributed in nature.[1] Both natural and synthetic derivatives have been reported to exhibit biological, pharmacological and biocidal properties.[1,2] The antioxidant capacity deserves a special mention and the structure-activity studies indicate that a substitution pattern that includes the presence of phenolic and/or catecholic moieties is highly important.[3]

Inspired by the chemical structures of natural products we developed new routes for the synthesis of the quite rare family of 2-styrylchromones 1 and challenged us to design novel bioactive derivatives.[4] The reactivity of 3-bromo-2-styrylchromones was explored for the synthesis of unique examples of 2,3-diarylxanthones 2.[5] Moreover, we focus our attention in the potential value of the hydroxylated 2-styrylchromones and –xanthones in order to establish structure-antioxidant activity relationships.[6]

Highlights of our efforts towards the synthesis, spectroscopic characterization and antioxidant profile of chromones 1 and xanthones 2 and of all the intermediates will be presented and discussed.

O

O

O

O

R2

R1

R6

R5

1 2

R3

R4

Acknowledgments: Thanks are due to the University of Aveiro, FCT and FEDER for funding the Organic Chemistry Research Unit. (project PEst-C/QUI/UI0062/2011). References [1] (a) Ellis, G. P. In Chromenes, Chromanones and Chromones 1977, John Wiley & Sons. (b) El-Seedi, H.

R.; El-Ghorab, D. M. H.; El-Barbary, M. A.; Zayed, M. F.; Göransson, U.; Larsson, S.; Verpoorte, R. Curr. Med. Chem. 2009, 16, 25813.

[2] Pinto, M. M. M.; Sousa, M. E.; Nascimento, M. S. J. Curr. Med. Chem. 2005, 12, 2517. [3] Gomes, A.; Freitas, M.; Fernandes, E; Lima, J. L. F. C. Mini-Rev. Med Chem. 2010, 10, 1. [4] Silva, A. M. S.; Pinto, D. C. G. A.; Cavaleiro, J. A. S.; Levai, A.; Patonay, T. Arkivoc 2004 (vii), 106. [5] Santos, C. M. M.; Silva, A. M. S.; Cavaleiro, J. A. S. Eur. J. Org. Chem. 2009, 2642. [6] (a) Santos, C. M. M.; Freitas, M.; Ribeiro, D.; Gomes, A.; Silva, A. M. S.; Cavaleiro, J. A. S.; Fernandes,

E. Bioorg. Med. Chem. 2010, 18, 6776. (b) Gomes, A; Neuwirth, O.; Freitas, M.; Couto, D.; Ribeiro, D.; Figueiredo, A. G. P. R.; Silva, A. M. S.; Seixas, R. S. G. R.; Pinto, D. C. G. A.; Tome, A. C.; Cavaleiro, J. A. S.; Fernandes, E.; Lima, J. L. F. C. Bioorg. Med. Chem., 2009, 17, 7218. (c) Gomes, A.; Fernandes, E.; Garcia, M. B. Q.; Silva, A. M. S.; Pinto, D. C. G. A; Santos, C. M. M.; Cavaleiro, J. A. S.; Lima, J. L. F. C. Bioorg. Med. Chem., 2008, 16, 7939. (d) Gomes, A.; Fernandes, E.; Silva, A. M. S.; Santos, C. M. M.; Pinto, D. C. G. A; Cavaleiro, J. A. S.; Lima, J. L. F. C. Bioorg. Med. Chem., 2007, 15, 6027.

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Development of new tetrapyrrolic macrocycles with potential application in medicine

Maria G. P. M. S. Neves

Department of Chemistry & QOPNA, University of Aveiro, 3810-193 Aveiro, Portugal

Porphyrins and analogues are widely distributed in Nature, playing essential roles in

vital processes such as photosynthesis, oxygen transport and storage. In the last decades many research groups have pointed out that these tetrapyrrolic macrocycles show unique features to be used in several applications like medicine, oxidative catalysis, as biomimetic model systems of primary processes of photosynthesis, as supramolecular units, including molecular recognition in chemical receptors and sensors and as materials for nanosciences. Most of the applications of these compounds in medicine are based on their use as photosensitizers in photodynamic therapy (PDT) of oncological and non-oncological diseases. The search for new tetrapyrrolic macrocycles with adequate features for specific applications has become the target of several work programs namely of the group of Aveiro. In here we will consider some work related with the functionalization of readily available tetrapyrrolic macrocycles (e.g. meso-tetraarylporphyrins or meso-triarylcorroles), aiming to improve their properties to be used either as photosensitizers or as chemosensors. [1-6]

Acknowledgments: Thanks are due to all the co-authors that contributed to the work presented here and to the University of Aveiro, “Fundação para a Ciência e a Tecnologia”-(FCT) and POCI 2010 (FEDER) for funding the Organic Chemistry Research Unit (Project PEst-C/QUI/UI0062/2011) and the Portuguese National NMR Network supported by funds from FCT. References

[1] Cavaleiro J.A.S., Tomé A.C., Neves M.G.P.M.S., Handbook of Porphyrin Science, Kadish K.M., Smith K.M., Guilard R., Eds., World Scientific Publishing Company Co., Singapore, 2010, vol. 2, pp 193.

[2] Carvalho C.M.B., Neves M.G.P.M.S., Tomé A.C., Almeida Paz F. A., Silva A.M.S., Cavaleiro J.A. S., Organic Lett., 2011, 13, 130.

[3] Moura N.M.M., Faustino M.A.F., Neves M.G.P.M.S., Almeida Paz F.A., Silva A.M.S., Tomé A.C., Cavaleiro J.A. S., Chem. Commun., 2012, 48 (49), 6142.

[4] Moura, N.M.M., Faustino, M.A.F., Neves, M.G.P.M.S., Tomé, A.C., Rakib, E.M., Hannioui, A., Mojahidi, S., Hackbarth, S., Röder, B., Almeida Paz, F.A., Silva, A.M.S., Cavaleiro, J.A.S. Tetrahedron, 2012, 68 (39), 8181.

[5] Costa, D.C.S., Gomes, M.C., Faustino, M.A.F., Neves, M. G. P.M.S., Cunha, Â., Cavaleiro, J.A.S., Almeida, A., Tomé J.P.C. Photochemistry Photobiology Science, 2012, DOI: 10.1039/C2PP25113B

[6] Almeida A., Cunha Â, Faustino M.A.F., Tomé A.C., Neves M.G.P.M.S., Photodynamic Inactivation of Microbial Pathogens: Medical Environmental Applications, Hamblin M.R., Jori G., Eds., Cambridge: Royal Society of Chemistrty. Pp423.

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Drugs, nutraceuticals and cosmeticals from marine sources

Anake Kijjoa

Instituto de Ciências Biomédicas de Abel Salazar and Centro Interdisciplinar de Investigação Marinha e Ambiental (CIIMAR), Universidade do Porto, Rua de Jorge Viterbo 228, 4050-313 Porto, Portugal

Marine Biotechnology, which involves marine bioresources, either as the source or

the target of biotechnology applications, is fast becoming an important component of the global biotechnology sector. The global market for Marine Biotechnology products and processes is currently estimated at € 2.8 billion (2010) with a cumulative annual growth rate of 4.5%. Less conservative estimates predict an annual growth in the sector of up to 10-12% in the coming years, revealing the huge potential and high expectations for further development of the Marine Biotechnology sector at a global scale.[1]

Because of the physical and chemical conditions in the marine environment, almost every class of marine organism possesses the capacity to produce a variety of molecules with unique structural features These molecules offer an unmatched chemical diversity and structural complexity, together with a biological potency and selectivity.[2] This is due, in large part, to the increased recognition of marine organisms as a source of bioactive compounds with pharmaceutical applications or other economically useful properties. The fact that marine resources are still largely unexplored has inspired many scientist to intensify their effort by using novel technologies to overcome the inherent problems in discovering compounds which may have potential for further development as pharmaceuticals or as functional products such as cosmetics, nutritional supplements and functional foods.

In recent years, the chemistry of natural products derived from marine organisms has become the focus of a much greater research effort. Currently, although there are around 15 marine natural products in various phases of clinical development, with more on the way and several products on the market, there is also great interest in marine-derived products as nutraceuticals and cosmeticals because of their beneficial effects on human health. Acknowledgments: This work was partially supported by the project PEst-C/MAR/LA0015/2011 funded by Fundação para a Ciência e a Tecnologia (FCT) and the European Regional Development Fund (ERDF) through the COMPETE - Operational Competitiveness Programme as part of the National Strategic Reference Framework. References [1] Bǿrrensen, T.; Boyen, C.; Dobson, A.; Höfle, M.; Ianora, A.; Jaspars, M.; Kijjoa, A.; Olafsen, J.;

Querellou, J.; Rigos , G.; Wijffels, R. Marine Board-ESF Position Paper 15, 2010, 1-91. [2] Kijjoa, A.; Sawangwong, P. Marine Drugs 2004, 2, 73-82.

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Design, synthesis, and biological evaluation of SIRT modulators

Antonello Mai

Department of Drug Chemistry and Technologies, Sapienza University of Rome, Rome, Italy

NAD+-dependent histone deacetylases (sirtuins, SIRT1-7) have emerged as potential therapeutic targets for treatment of human illnesses such as cancer, metabolic, cardiovascular and neurodegenerative diseases. In our lab, several chemically different series of sirtuin inhibitors (SIRTi) have been identified and will be further developed.

The first series comprises some sirtinol analogues, obtained by replacement of the benzamide linkage of the prototype with other bioisosteric groups. Condensation of 2-hydroxy-1-naphthaldehyde with the appropriate N-(2-, 3-, and 4-aminophenyl)-2-phenylpropanamides, or 2-, 3-, and 4-amino-N-(1-phenylethyl)benzenesulfonamides, or 2-, 3-, and 4-(2-phenylpropylthio)-, -(2-phenylpropylsulfinyl)-, or –(2-phenylpropylsulfonyl)- -anilines led to the desired compounds. Such derivatives showed higher apoptosis induction and/or higher cytodifferentiation than sirtinol in human leukemia U937 cells. Starting from cambinol, a SIRTi reported as highly active against the Burkitt lymphoma, we designed some (thio)barbituric acid analogues (BDF4s) related to cambinol to evaluate as SIRTi and anticancer agents. The BDF4 prototype, MC2141, obtained by condensation and intramolecular dehydratation of N-phenylbarbituric acid and 2-hydroxy-1-naphthaldehyde, displayed in U937 cells higher apoptosis induction than cambinol and showed antiproliferative effects against a panel of cancer cells. More recently we devoted our attention to AGK-2, a SIRT2-selective inhibitor useful in a Parkinson disease model. We designed and synthesized some AGK-2 analogues, called acylpyrrolyliden-cyanacetamides (APCs) bearing an acylpyrrole moiety instead of the phenylfuran group of AGK-2, and we started to study the effect of substitution at the pyrrole as well as cyano and/or quinoline level on the SIRT inhibiting activity.

In contrast to the number of SIRT inhibitors, only few SIRT1 activators are known. Here, we rationalized the potential of the previously unexplored dihydropyridine scaffold to develop sirtuin ligands by preparing a series of 1,4-dihydropyridine-based (DHP) derivatives through one-pot reaction with arylaldehyde, ethyl propiolate, and the appropriate alkyl/arylamines. Assessment of their SIRT1-3 deacetylase activities revealed the importance of the substituent at the N1 position of the dihydropyridine structure on sirtuin activity. Introduction of cyclopropyl, phenyl, or phenylet hyl groups at N1 conferred non-selective SIRT1 and SIRT2 inhibition activity, while a benzyl group at N1 conferred potent SIRT1 and -2 activation. Senescence assays performed on hMSC, and mitochondrial function studies conducted with murine C2C12 myoblasts confirmed the compounds’ novel and unique SIRT-activating properties.

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Redox chemistry of sulfur compounds and its biological relevance

Richard S. Glass

Department of Chemistry and Biochemistry, The University of Arizona, Tucson, AZ 85721, U .S.A Dedicated to the memory of Prof. Dr. Klaus-Dieter Asmus

The redox chemistry of thioethers exemplified by the side-chain of methionine are

important biological and pharmacologically. Long-range electron-transfer in redox proteins may occur by a “hopping” mechanism through a methionine side-chain[1] or alternating sulfur π-aromatic amino acid side chains.[2] Methionine residues in proteins may serve a protecting role as antioxidants.[3] They are oxidized to sulfoxide moieties which are reduced in a reaction catalyzed by methionine reductase.[4] Alternatively, they can serve a deleterious role by generating reactive oxygen species as suggested for the pathogenesis of Alzheimer’s disease.[5]

To validate the chemistry proposed for methionine residues in proteins and peptides we have synthesized conformationally constrained molecules in which thioether moieties are juxtaposed with amide or aromatic residues and studied their redox chemistry. Thus amide 1, in which the thioether and amide moieties are held close together, have been synthesized and characterized. Electrochemical studies show that such compounds undergo oxidation at a potential over 0.5 V less anodic than analog 2 in which through-space interaction is precluded.[6] Furthermore, time resolved one-electron oxidation of 1 , using pulse radiolysis, shows the formation of a novel 2c, 3e S, O-bonded intermediate. Similar results are found for 3 in which S-π through space interaction lowers the oxidation potential by almost 0.5 V.[7] Multicenter interaction has been found for m-terphenyl thioether 4 involving two arenes and one thioether moiety and 5 involving three arenes and two thioether moieties.[8] These compounds may prove to be of interest as pharmaceutical agents as well owing to their redox properties.

Acknowledgements: The author gratefully acknowledges support of this research by the U.S. National Science Foundation through Grants CHE-0527003 and CHE-0956581. References [1] (a) Wang, M.; Gao, J.; Müller, P.; Giese, B. Angew. Chem. Int. Ed., 2009, 48, 1. (b) Giese, B.; Wang, M.; Gao

J. Stoltz, M.; Müller, P.; Graber, M. J.Org. Chem. 2009, 74, 3621. (c) Cordes, M.; Giese, B. Chem. Soc. Rev. 2009, 38, 892.

[2] Morgan, R.S.; Tabsch, C.E.; Gushard, R.H.; McAdon, J.M.; Warme, P.K. J. Peptide Protein Res. 1978, 11, 209.

[3] (a) Stadtman, E.R. Arch. Biochem. Biophys. 2004, 423, 2. (b) Luo, S.; Levine, R.L. FASEB J. 2009, 23, 464. [4] Boschi-Mueller, S.; Grand, A.; Branlant, G. Arch. Biochem. Biophys. 2008, 474, 266. [5] Schöneich, C. Biochem. Biophys. Acta. 2005, 1703, 111. [6] (a) Glass, R.S.; Hug, G.L; Schöneich, C.; Wilson, G.S.; Kuznetsova, L.; Lee, T.-M.; Ammam, M.; Lorance,

E.; Nauser, T.; Nichol, G.S.; Yamamoto, T. J. Am. Chem. Soc. 2009, 131, 13791. (b) Glass, R.S.; Schöneich, C.; Wilson, G.S.; Nauser, T.; Yamamoto, T.; Lorance, E.; Nichol, G.S.; Ammam, M. Org. Lett. 2011, 13, 2837.

[7] Chung, W.J.; Ammam, M.; Gruhn, N.E.; Nichol, G.S.; Singh, W.P.; Wilson, G.S.; Glass, R.S. Org. Lett. 2009, 11, 397.

[8] Zakai. U.T.; Bloch-Mechkour, A.; Jacobsen, N.; Abrell, L.; Lin, G.; Nichol, G.S.; Bally, T.; Glass, R.S. J. Org. Chem. 2010, 75, 836.

Invited Lectures

INVITED LECTURES

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Control of plant diseases with natural products: allicin from garlic as an example

Alan J. Slusarenko

Department of Plant Physiology, RWTH Aachen University, 52056 Aachen, Germany

The volatile antimicrobial substance allicin (diallylthiosulfinate) is produced in garlic

when the tissues are damaged and the substrate alliin (S-allyl-L-cysteine sulfoxide) mixes with the enzyme alliin-lyase (E.C.4.4.1.4).[1] Allicin is an example of a reactive sulfur species (RSS)[2] and is readily membrane-permeable and a pro-oxidant which undergoes thiol-disulfide exchange reactions with free thiol groups in proteins and it was suggested that inactivation of thiol-containing enzymes was the basis of its antimicrobial action.

S

O

C

H

COOH

NH2

S

O

S2 H2O + 2pyruvate + 2NH3

alliinase

alliin allicin We tested the effectiveness of garlic juice containing allicin against a range of plant

pathogenic organisms in vitro and in planta. Disease reduction in planta was shown for Magnaporthe grisea-infected rice, Hyaloperonospora parasitica-infected Arabidopsis thaliana and Phytophthora infestans-infected potato tubers and tomato plants. We investigated cellular mechanism of action of allicin using Saccharomyces cerevisiae as a model organism.

SHS

O

SS

S

allicinthiol

2R + 2R

mixed disulphide

+ H2O

We measured changes in the absolute concentrations of reduced and oxidized

glutathione after allicin treatment. Calculation of the cellular redox potential using the Nernst equation showed that the cellular electrochemical potential was shifted from a highly reduced to a more oxidized state. Changes in the electrochemical potential of the GSSG/GSH couple, quantitatively the most important redox buffer in the cell, correlate with the biological status of the cell and it is predicted that the allicin-induced redox shift shunts the cells into apoptosis.[3] Yeast caspase was activated in cells treated with pure synthetic allicin or garlic juice. Furthermore, cells were protected by expressing the antiapoptotic BclxL or by inhibiting the endogenous yeast AIF (apoptosis inducing factor) gene; confirming that allicin kills cells by acting as a redox toxin which shunts cells into genetically programmed apoptosis or, at higher dosages, shifts the redox state in the cells so much that they simply necrose. References [1] Cavallito CJ, Bailey HJ. (1944) J. Am. Chem. Soc. 66:1950-51. [2] Jacob C, Anwar A (2008). Physiologia Plantarum 133(3): 469-480 [3] Gruhlke et al. (2010) Free Radical Biol. Med. 49, 1916-1924

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Natural compounds as inhibitors of the 10 hallmarks of cancer

Marc Diedericha,b

a Laboratoire de Biologie Moléculaire et Cellulaire du Cancer, Hôpital Kirchberg, 9 rue Edward Steichen, L-2540 Luxembourg, Grand Duchy of Luxembourg; b Department of Pharmacy, College of Pharmacy, Seoul

National University, Seoul, 151-742, KOREA

Cancer is one of the most deadly diseases in the world. Although advances in the field

of chemo-preventive and therapeutic medicine have been made regularly over the last ten years, the search for novel anticancer treatments continues. In this field, the natural environment, with its rich variety of organisms, is a largely untapped source of novel compounds with potent antitumor activity. We focus here on selected compounds that act on the eight major hallmarks of cancer including self-sufficiency in growth signals, insensitivity to anti-growth signals, evasion of apoptosis, limitless replication, sustained angiogenesis and tissue invasion, metastasis, altered cellular metabolism and the evasion of immune destruction. Moreover, we identify compounds that interfere with the two enabling characteristics coined by Hanahan and Weinberg recently, namely inflammation and genome instability.

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Redox signaling via the cellular thiolstat - the special relationship in group 16

Claus Jacob, Torsten Burkholz, Thomas Schneider, Brigitte Czepukojc, Uma Viswanathan

Division of Bioorganic Chemistry, School of Pharmacy, Saarland University, Building B 2.1, Campus, D-66123

Saarbruecken, Germany, c.jacob@mx.uni-saarland.de

Research conducted during the last decade has indicated that the mammalian cell contains a wide range of redox sensitive cysteine proteins, whose oxidation (S-thiolation) state is critical to function and activity. Together, these proteins form a complex and sophisticated network which enables the cell to sense subtle redox changes and to respond in a rapid, measured, differentiated and vastly reversible manner. This ‘cellular thiolstat’ is involved in numerous cellular signalling pathways which affect, for instance, proliferation, antioxidant defence, differentiation and cell death.

The sophistication of the thiolstat is based on three major ‘chemical specialities’ related to the redox chemistry of sulfur: (a) the redox behaviour of the thiol group of cysteine is highly diverse - it involves numerous oxidation states accessible by various redox mechanisms, (b) individual cysteine residues in proteins differ dramatically as far as their redox sensitivity and accessibility for oxidation is concerned, (c) the modification of individual cysteine residues may trigger a wide and diverse range of cellular responses, (d) most cysteine modifications are reversible.

Unlike other posttranslational protein modifications involved in cellular signalling, S-thiolation (and cysteine overoxidation) are not only enzyme catalyzed, but can also be affected by exogenous agents. Here, redox modulators based on sulfur, selenium and tellurium bear great promise. Many of these agents are highly specific for cysteine residues and hence are able to modify specific cellular proteins, such as β-tubulin, with high efficiency yet also with considerable precision. Such redox modulating agents can trigger specific cellular processes, such as apoptosis, in cells with a particular ‘biochemical signature’, such as cancer cells. Ultimately, this combination of efficiency and selectivity associated with various cysteine-specific agents may be exploited in the field of drug design.

References [1] C. Jacob, Biochem. Soc. Trans., 39, 1247-1253 (2011). [2] M. Doering, L.A. Ba, N. Lilienthal, C. Nicco, C. Scherer, M. Abbas, A.A. Peer Zada, R. Coriat, T.

Burkholz, L. Wessjohann, M. Diederich, F. Batteux, M. Herling and C. Jacob, J. Med. Chem., 53, 6954-6963 (2010).

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Bacillithiol: The emerging redox properties and functions of the glutathione surrogate in many Gram positive bacteria

Chris Hamilton

School of Pharmacy, University of East Anglia, Norwich Research Park, Norwich NR4 7TJ, UK

BS

BSH

BSSB[O]

(H2O2,NO...)

GlcN-Mal

Protein

S SB

bacillithiolation

bacilliredoxins

BSSBreductase

Protein

SH

Cys

E+

CyS

xenobioticdetoxif ication

HO O

HO

OHN

OH

CO2H

CO2H

O

NH2

HS

Bacillithiol (BSH)

Low molecular weight thiols such as glutathione (GSH) (eukaryotes, Gram negative bacteria) and mycothiol (MSH) (actinomycetes) play a critical role in resistance and regulation of oxidative stress. Bacillithiol (BSH) was recently identified as a unique low molecular thiol amongst many low G+C Gram positive bacteria (eg. B. subtilis, B. anthracis, S.aureus and B. cereus), which do not produce GSH or MSH.[1] BSH deficient mutants display increased sensitivity to toxic oxidants, metal ions and some electrophilic antibiotics.[2] Emerging genetic and biochemical evidence already suggests that BSH serves analogous biochemical functions to those already known for GSH and MSH in other micro organisms.[2-

4] We have recently completed a total chemical synthesis of BSH and its symmetrical disulfide (BSSB)[4] as well as polyclonal antibodies able to detect bacillithiolated proteins. We are now using these materials to establish BSH redox properties as well as the identity and function of BSH-dependent redox proteins. Recent progress and discoveries within this emerging arena of low molecular weight thiol biochemistry will be discussed. References [1] Nat. Chem. Biol. 2009, 5, 625. [2] Proc. Nat. Acad. Sci. USA, 2010, 107, 6482. [3] Antiox. Redox Signal. 2012, (in press) DOI:10.1089/ars.2012.4686. [4] Angew. Chem. Int. Ed. 2011, 50, 7101.

Oral and Poster Communications

ORAL AND POSTER COMMUNICATIONS

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Garlic in agriculture - developments and challenges

Awais Anwar and Murree Groom

ECOspray Limited, Grange Farm Hilborough, Thetford IP26 5BT, Norfolk, UK

The intensification of agriculture to fulfil global food needs has increased the number of insect, fungal and nematode pest species attacking crops and increased their simultaneous resistance to many crop protection products. The combined effect of these two factors increases the risk of lost production, balanced to some extent by increased cycles of intensification that are now considered un-sustainable.

Using plant extracts as crop protection products is an emerging technique because of the toxicity and residue problems affecting the food chain from synthetic products. The literature is extensive concerning the possible use of plants as crude or refined extracts[1-3] in various areas of crop protection (insects, fungi, nematodes, bacteria, weeds). It is now mandatory though to attribute the efficacy of botanicals to specific identified constituent compound(s) in order to delineate the mechanisms of bioactivity, biologically and biochemically, and to fully exploit the therapeutic potential of extracts.[4]

The history, mystery and magic of the garlic plant has seduced people from all over the world for generations. Garlic extracts and its active compounds (allylic polysulfides) have been studied widely to control pests and human disease around the world.[5-7] In the agriculture arena ECOspray has been involved in developing different formulations to control pests with a special focus on nematicides. Various formulations have been tested in field and green houses conditions, producing efficacy comparable to highly toxic synthetic alternatives.

In recent years ECOspray has developed formulations in areas of bird repellency, animal feed inclusion, sports turfs and general nematicide for many vegetables all based on the environmentally friendly active molecules found in garlic.

Crop protection products are some of the most thoroughly tested and regulated chemicals in the world. The process for bringing new pesticides to the market is quite challenging.

A concerted effort in formulation development for bio-pesticides by multi-disciplinary teams is still required to optimize bio-pesticide yield, efficacy, storage stability and delivery and thus enable this technology to evolve further and make a significant contribution to meeting today's agricultural and societal demands for safe and sustainable food production. References [1] G. T. Brooks, Pesticide Science, 1998, 52, 303-304. [2] O. Koul and G. S. Dhaliwal, Phytochemical Biopesticides, Harwood Academic, 2001. [3] L. G. Copping, Pesticide Science, 1999, 55, 756-757. [4] M. Akhtar and I. Mahmood, Bioresource Technology, 1994, 48, 189-201. [5] C. Jacob and A. Anwar, in Chemoprevention of Cancer and DNA Damage by Dietary Factors, ed. S.

Knasmueller, D. DeMarini, I. Johnson and C. Gerhaeuser, Wiley-VCH Verlag GmbH & Co., Weinheim, 2009, pp. 663-684.

[6] C. Jacob, Natural Product Reports, 2006, 23, 851-863. [7] C. Cerella, C. Scherer, S. Cristofanon, E. Henry, A. Anwar, C. Busch, M. Montenarh, M. Dicato, C.

Jacob and M. Diederich, Apoptosis, 2009, 14, 641-654.

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Effective formulations of garlic in agriculture

Awais Anwar,a Miriam Arbach,b Murree Groom,a Chris Hamiltonb

aECOspray Limited, Grange Farm Hilborough, IP26 5BT, Norfolk, UK; bSchool of Chemistry, University of East Anglia, Norwich, NR4 7TJ, UK.

Clearly the formation and the physical and biochemical properties of garlic’s

bioactive organosulfur metabolites are very complex.[1] Such complexities create many challenges that need to be addressed in their formulation, application and regulation within an agricultural setting. However, such challenges are worthwhile pursuing, because such food-derived bioactive molecules can provide much more desirable alternatives to many of the environmentally persistent agrochemical pesticides that are currently in use.

Recent initiatives by the pesticide regulatory departments of European and North American governments have stimulated renewed interest in bio-pesticide technologies to replace toxic synthetic pesticides with more benign natural products. Much progress has been made recently with bringing botanical bio-pesticides to the market and the first well researched examples of these products are starting to enter significant segments the EU crop protection market.

The extensively studied sulfur compounds derived from garlic[2] represent excellent candidates for green nematicides or insecticides. One of the nematicides developed by ECOspray, NEMguard®, is capable of delivering efficacy comparable to synthetic products such as Temik (aldicarb), Vydate (oxamyl) and Nemacur, all of which are under intense international regulatory scrutiny for adverse ecotoxicity and residues in treated crops. References [1] E. Block, Garlic and other alliums: the lore and the science, RSC Pub., Cambridge, UK, 2010. [2] A. Anwar, T. Burkholz, C. Scherer, M. Abbas, C.-M. Lehr, M. Diederich and C. Jacob, Journal of Sulfur

Chemistry, 2008, 29, 251-268.

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Synthesis and biological activity of 1H-naphtho[1,2-c]chromene-1,4,5-trione and benzo[i]phenanthridine-1,4,5(6H)-trione derivatives

Z. Xu,a,b,c S. Valente,a C. Jacob,b P. Meiser,c G. Kirscha

aLaboratory of Molecular Engineering and Pharmacological Biochemistry, Jean Barriol Institute, University

of Lorraine, 1 Boulevard Arago, 57070 Metz, France; bDivision of Bioorganic Chemistry, School of Pharmacy, Saarland University, D-66123 Saarbrücken, Germany; cURSAPHARM Arzneimittel GmbH, D-

66129 Saarbrücken, Germany.

2H-1-benzopyran-2-one (coumarin) is present in a large number of natural products as part of their structure.[1] It has been one of the most valuable sources of novel compounds with diverse biological activity.[2] Construction of novel coumarin-based ring systems would also lead to the development of promising templates for drug discovery. Recently, we have descried the novel coumarin-based polycycles synthetic by [4+2] thermal Diels-Alder cycloaddition reaction, and their CDC 25 phosphatases activity.[3] Among these novel compounds, the 1H-naphtho[1,2-c]chromene-1,4,5-trione showed an interesting inhibitory activity in vitro against all of three CDC25 phosphatases.

Therefore, a series of new 1H-naphtho[1,2-c]chromene-1,4,5-trione and benzo[i]phenanthridine-1,4,5(6H)-trione derivatives were synthesized by [4+2] thermal Diels-Alder cycloaddition reaction. We applied the Heck reaction to 4-bromocoumarin 1 and 4-bromoqunlin 2 using Pd2dba3/DPPF as catalytic system, we obtained selectively 4-(2-butoxyvinyl)coumarin (or quinolin) 3. 1H-naphtho[1,2-c]chromene-1,4,5-trione and benzo[i]phenanthridine-1,4,5(6H)-trione derivatives 4 were synthesized by (4π+2π) cycloaddition reaction (Diels-Alder reaction). All the compounds were tested in vitro for their antiproliferative activity against tumor cell lines and inhibitory activity against CDC 25A, B and C phosphatases.

Acknowledgments: Thanks for funding by the European Community’s Seventh Framework Programme (FP7/2007-2013) under grant agreement 215009 RedCat. References [1] Kostova, I. Curr. Med. Chem. Anticancer Agents 2005, 5, 29–46. [2] Starčević, S.; Brožič, P.; Turk, S.; Cesar, J.; Lanišnik Rižner, T. and Gobec, S. J. Med. Chem. 2011, 54,

248–261. [3] Valente, S.; Bana, E.; Viry, E.; Bagrel, D. and Kirsch, G. Bioorg. Med. Chem. Lett. 2010, 20, 5827-30.

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Michael addition on chromone-3-carboxylic acid: a one-pot tandem reaction towards novel polysubstituted -chromones, -chromanones, and -

flavones

Oualid Talhi,a Jana Panning,b Clemens Zwergel,c Diana C. G. A. Pinto,a Claus Jacob,b Gilbert Kirsch,c Artur M. S. Silvaa

aDepartment of Chemistry and QOPNA, University of Aveiro, 3810-193 Aveiro, Portugal; bSchool of

Pharmacy Building B 2.1., Room 1.13 Campus 66123 Saarbrucken, Germany; cLaboratoire d’Ingénierie Moléculaire et Biochimie Pharmacologique1, boulevard Arago 57070 Metz France

Benzopyran-4-one derivatives is a large family of compounds with a widespread

importance in chemistry and biology. A huge amount of investigations are considering the broad biological activities inherent in the benzopyran-4-one-based compounds.[1] In the current work, basic organocatalysis has been used for a one-pot tandem synthesis of novel benzopyran-4-one derivatives, including chromones, chromanones, flavones and 2-styrylchromones. The tandem process involves four steps, Michael-addition / decarboxylation / pyran-4-one-ring-opening of chromone-3-carboxylic acid 1 upon reacting with 1,3-diketone (or 1,3-dicarbonyl compounds) 2 which undergoes a final chromone- and / or chromanone- ring closure step in case where R1 is an ortho-hydroxyphenyl group. Several polyphenolic scaffolds 3-6 sharing the chalcone-type moiety or bearing chiral core structures, have been produced depending on the structural patterns of the used 1,3-dicarbonyl active methylenes 2. Mechanistic, structural and stereochemical studies of the novel structures will be deeply discussed based on spectral data recorded from 1D and 2D NMR experiments.

O

O

COOH

R1

O

R2 O

Michael addtion

O

R2 O

O

R1

O

O

OHR2

R1

O

OH

O

O

O

OH

R21

2

3

5

4

6

O

O

O

HO

OH

Carbonnucleophile

Chromone-ring closure Chromanone-ring closureR1 = ortho-hydroxyphenyl

**

*R

R

R

Acknowledgments: Thanks are due to the University of Aveiro, FCT and FEDER for funding the Organic Chemistry Research Unit and the Portuguese National NMR Network (RNRMN). The authors also thank the (European Community’s) Seventh Framework Programme (FP7/2007-20139] under grant agreement nº 215009) for the financial support. References [1] Harborne, J.B.; Grayer, R.J. The Flavonoids, Advances in research since 1986, J.B. Harborne ed.;

Chapman and Hall: London, 1993.

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Xanthohumol from hops activates the cytoprotective Keap1-Nrf2 pathway through its enone site

Lidia Brodziak-Jarosz,a,b Sabine Amslinger,c C. Roland Wolf,d Clarissa Gerhäuser,a Tobias

Dickb

aDivision of Epigenomics and Cancer Risk Factors, bDivision of Redox Regulation, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69120 Heidelberg, Germany; cDepartment of Organic

Chemistry, University of Regensburg, Universitaetsstrasse 31, 93053 Regensburg, Germany; dCancer Research UK Molecular Pharmacology Unit, Ninewells Hospital and Medical School, University of Dundee,

Dundee DD1 9SY, Scotland, United Kingdom

Xanthohumol (XN) is a prenylated chalcone from hops (Humulus lupulus L.), demonstrated to have anti-tumorigenic activities. XN may act, at least partially, by influencing cellular redox processes. In fact, XN has been identified recently as an inducer of mitochondrial superoxide generation. Moreover, XN has electrophilic properties due to its enone moiety (a Michael acceptor site), that may be attacked by nucleophilic thiols, thus potentially giving rise to covalent protein thiol modifications.

Our study addressed the question of whether the redox-modulating properties of XN play a role in the activation of signaling pathways contributing to the cell survival/cell death decision. The cytoprotective Keap1-Nrf2 pathway responds to both oxidants and electrophiles, and regulates expression of genes containing an antioxidant-response element (ARE) in the promoter region.

We analyzed activation of the Keap1-Nrf2 pathway in MCF7-derived AREc32 cells using an ARE-driven luciferase reporter assay. Treatment of cells with XN strongly induced reporter activity, thus indicating activation of the Keap1-Nrf2 pathway. Chalcone, which contains a similar enone site, showed a similar potency in inducing an Nrf2 response. In contrast, treatment of cells with tetrahydroxanthohumol (TX), a derivative of XN lacking the electrophilic enone site, did not trigger the Keap1-Nrf2 pathway, suggesting that the enone site is essential for XN-mediated Keap1-Nrf2 activation.

Considering the possibility that the enone site of XN forms covalent adducts with reactive protein thiols, we synthesized an alkynylated analogue of XN to label, enrich and identify such adducts by click chemistry and mass spectrometry. We identified as many as 150 protein targets of XN derivative, including Keap1. This result suggests a pleiotropic effect of XN on multiple pathways and processes involving reactive cysteines. The interaction of XN-alkyne with Keap1 was additionally confirmed be western blotting, thus providing in situ evidence of direct alkylation of Keap1 cysteine(s) by XN derivative, what may be an important mechanism of Nrf2 activation by XN.

Xanthohumol structure Acknowledgments: Thanks are due to the (European Community’s) Seventh Framework Programme (FP7/2007-20139] under grant agreement nº 215009) for the financial support.

Enone site

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Synthesis and biological evaluation of aurones and chalcones from coumarins and chromones

Clemens Zwergel a, Sergio Valentea,b, Gilbert Kirscha

aLaboratoire d’Ingénierie Moléculaire et Biochimie Pharmacologique, Institut Jean Barriol, Université de Lorraine, 1 Boulevard Arago, 57070 Metz, France ; bDipartimento di Chimica e Tecnologie del Farmaco,

Istituto Pasteur–Fondazione Cenci Bolognetti, Università di Roma “La Sapienza,” 00185 Roma, Italy

Coumarin and Chromone based aurones[1]: Not widely distributed in nature, aurones, (Z)-2-benzylidene-benzofuran-3(2H)-ones, are one of the less common and lesser-known representatives of a flavonoid subclass. This is probably the reason why they have received little attention in comparison to the structurally similar and widely investigated flavones and isoflavones. Nevertheless they also exhibit a strong and broad variety of biological activities we recently summarized in our review[1] For example they have been described as antifungal agents, as insect antifeedant agents, as inhibitors of tyrosinase, and as antioxidants. Our idea was now to combine the benzofuranone 9 part of a classical aurone with two well-known structures: the coumarin 7 and the chromone 8 which proved several interesting biological activities such as antimicrobial, antiviral, anticancer, anti-inflammatory, antioxidant.

The novel compounds are currently tested in a redox assay as well as tested in leukemia cells K562.

Coumarin based chalcones as potential NFkB pathyway inhibitors [2]: NF-κB pathway controls the expression of several targets genes involved in apoptosis, cell proliferation, innate and acquired immunity and inflammation. Aberrant activation of the NF-κB pathway is known as to be involved in many diseases and cancers. A promising approach in the fight against cancer is the discovery of new NF-κB inhibitors. Based on known inhibitors like chalcones and curcurmin (2 or 3) we developed analogues containing

the coumarin moiety. In summary, we described novel derivatives of coumarins and chromones. Among these novel heterocycles we started the biological evaluation in vitro (redox assay) and in vivo (leukemia cells K562, NFkB pathway activation).

Acknowledgments: I thank all the RedCat Partners and especially Prof G. Kirsch, Prof C. Jacob, Prof A. Mai, Prof L. Altucci, Dr M. Diederich and Dr S. Valente for their continuous support and help in this work. This work is funded by the European Community, 7th Framework Programme under grant agreement No 215009 (RedCat Marie Curie ITN Network). References [1] Zwergel, C.; Gaascht, F.; Valente, S.; Diederich, M.; Bagrel, D.; Kirsch, G.; Aurones: Interesting natural

and synthetic compounds with emerging biological potential; Nat. Prod. Commun. 2012, 7, 3, 389-394. [2] Yadav, B.; Taurin, S.; Rosengren, RJ.; Schumacher, M.; Diederich, M.; Somers-Edgar, TJ.; Larsen, L.

Bioorg. Med. Chem. 2010, 18, 6701.

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Oxidative stress in bacteria: differential sensitivities to selected oxidizing agents

Ifeanyi D. Nwachukwu, Alan J. Slusarenko*

RWTH Aachen University, Department of Plant Physiology (BioIII), 52056 Aachen, Germany.

*To whom correspondence should be addressed: alan.slusarenko@bio3.rwth-aachen.de

Upon wounding, garlic releases the volatile phytoanticipin allicin (diallylthiosulfinate) when its alliin (S-allyl-L-cysteine sulfoxide) substrate is acted upon by the enzyme alliinase (alliin lyase). Allicin, arguably the most biologically active compound in freshly prepared garlic juice is credited with potent antimicrobial and pharmacological properties that have made garlic increasingly important in medicine and agriculture.

Like H2O2, allicin is thought to kill cells via an oxidative mechanism. Since allicin is a redox-toxin; we presumed that sensitivity to allicin might be correlated generally with oxidative stress resistance or susceptibility. Using an agar plate diffusion bioassay, we exposed a number of bacteria (E. coli and Pseudomonas spp.), including a highly allicin-resistant pseudomonad originally isolated from garlic bulbs, to both allicin and H2O2. Our results have shown that several of the pseudomonads vary naturally in their sensitivities to allicin and to H2O2. While there is a general correlation between oxidative stress resistance (H2O2) and resistance to allicin in certain Pseudomonas isolates tested, for some Pseudomonas isolates, differential sensitivities to allicin and H2O2 have instead been observed. This is also the case for the E. coli: the gshA mutant, deficient in some of the enzymes essential for glutathione conjugation,[1] is more susceptible to allicin than to H2O2, while the oxyR mutant, lacking OxyR, a transcriptional regulator of oxidative stress-induced genes,[2] is more resistant to allicin than to H2O2.

This suggests that while both allicin and H2O2 cause oxidative stress, their cellular effects are different. This may be related to allicin’s ability to enter into thiol-disulfide exchange reactions which is not the case for H2O2. Acknowledgement: The research leading to this work was supported by funding from the European Community's 7th Framework Programme [FP7/2007-2013] under grant agreement No: 215009. References [1] Kumar, C.; Igbaria, A.; D’autreaux, B.; Planson, A-G.; Junot, C.; Godat, E.; Bachhawat, A.K.;

Delaunay-Moisan, A.; Toledano, M.B. The EMBO Journal. 2011, 30 (10):2044-2056. [2] Kullik I.; Toledano, M.B.; Tartaglia, L.A.; Storz, G. Journal of Bacteriology. 1995, 177 (5): 1275–

1284.

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Nitric oxide and hydrogen sulfide cross-talk: regulation of bioavailability and cell signalling

Maria Letizia Lo Faro,a John More,b Mark E. Wood,c Jacqueline Whatmore,a Paul G.

Winyard,a Matthew Whitemana

aUniversity of Exeter Medical School, Exeter, UK; bBioProducts laboratory, Dagger Lane, Elstree,

UK;cSchool of Biosciences, University of Exeter, Exeter, UK

Nitric oxide (˙NO) and hydrogen sulfide (H2S) are two gasotransmitters with key roles in the regulation of the vascular system, both in physiological[1,2] and pathophysiological conditions.[3,4] The two gases share a number of different functions, for example they are both involved in vasodilation and protect the vasculature after oxidative insults (like ischaemia-reperfusion injury[5]). Currently there is increasing interest in investigating if and at what level ˙NO and H2S interact and affect each other’s signalling pathways.[6]

The aims of this work were: to study if H2S donor molecules recycle ˙NO from ˙NO-derived metabolites (e.g. nitrite, NO2

-, and nitrosothiols, RSNOs) and to examine the effect of these reactions in cellular systems relevant for the vascular functions (smooth muscle and microvascular endothelial cells). In addition we examined ˙NO-mediated protein modifications (on Cys residues) and their effect on protein functions.

H2S was able to induce production of ˙NO from both NO2- and RSNOs (as assessed

by electron paramagnetic resonance spectroscopy, EPR, and gas phase ozone-based chemiluminescence) (e.g. [˙NO] from NO2

-+H2S was 6.5±1.9 µM, n=4, EPR determination). These reactions affected ˙NO signalling, as evidenced by increased levels of cGMP production in smooth muscle cells (cGMP ELISA), and RSNO signalling, demonstrated by an increase in intracellular RSNO content in endothelial cells. S-nitrosated albumin (SNOA), as an example of a ˙NO-modified protein, was shown to protect endothelial cells from peroxide-induced oxidative stress to a greater extent than unmodified albumin (cell viability assessed by Sulforhodamine B staining).

In conclusion our results suggest that ˙NO and H2S are able to interact at the level of ˙NO-derived metabolites (NO2

- and RSNOs), resulting in the regulation of ˙NO levels. In addition ˙NO-modified albumin showed increased cytoprotective and anti-oxidant properties, suggesting novel clinical applications. Acknowledgments: This research received full funding from the European Community’s Seventh Framework programme (FP7/2007-2013 under grant agreement number 215009). References [1] Ignarro, L.J., et al., Endothelium-derived relaxing factor produced and released from artery and vein is

nitric oxide. Proc. Natl. Acad. Sci. U S A, 1987. 84(24): p. 9265-9. [2] Bhatia, M., Hydrogen sulfide as a vasodilator. IUBMB Life, 2005. 57(9): p. 603-6. [3] Huang, P.L., et al., Hypertension in mice lacking the gene for endothelial nitric oxide synthase. Nature,

1995. 377(6546): p. 239-42. [4] Chen, L., et al., Imbalance of endogenous homocysteine and hydrogen sulfide metabolic pathway in

essential hypertensive children. Chin. Med. J. (Engl), 2007. 120(5): p. 389-93. [5] Moody, B.F. and J.W. Calvert, Emergent role of gasotransmitters in ischemia-reperfusion injury. Med.

Gas Res., 2011. 1(1): p. 3. [6] Ali, M.Y., et al., Regulation of vascular nitric oxide in vitro and in vivo; a new role for endogenous

hydrogen sulphide? Br. J. Pharmacol., 2006. 149(6): p. 625-34.

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Effect of diallyl tetrasulfide on HCT116 colorectal cancer cells

N. E. B. Saidu, C. Jacob, M. Montenarh

Medical Biochemistry and Molecular Biology, University of the Saarland, Building 44, 66424 Homburg, Germany

Background: Diallyl polysulfanes have been shown to exert cell cycle arrest, anti-

tumour and anti-inflammatory activities in a variety of in vitro and in vivo models. Although diallyl polysulfanes cause oxidative stress, little is known about the underlying signaling cascades leading to antioxidant defense or apoptosis.

Methods: Cells were treated with DATTS at different concentrations and for different periods. Reactive oxygen species and thiol concentrations were determined by commercially available kits. The expression levels of signal molecules were determined by Western Blot analysis. A direct influence of Nrf2 on the promoter of HO-1 gene was determined by a luciferase assay with the StRE promoter element from the HO-1 gene.

Results: We found an immediate increase in the level of the superoxide anion radical O2

•- and hydrogen peroxide H2O2 and an overall thiol depletion. DATTS treatment of HCT116 cells also caused an up-regulation of phospho-eIF2α, nuclear Nrf2 and HO-1 protein levels in a time and concentration-dependent manner. Pre-treatment of cells with antioxidants significantly reduced the elevated expression levels of these proteins. A direct contribution of Nrf2 was shown by its interaction with the stress response element of the HO-1 promoter.

Conclusions: DATTS activates the ROS- eIF2α/Nrf2 HO-1 signaling cascades leading to the up-regulation of HO-1. However, this antioxidant defense is not sufficient to protect HCT116 cells from apoptosis.

General significance: This study shows for the first time a parallel but not equal activation of signaling pathways by DATTS with a competitive ultimate cellular outcome.

Keywords: reactive oxygen species (ROS), thiols, redox reagents, antioxidant pathway, apoptosis Acknowledgments: This work was supported by the European Community’s framework programme (FP7/2007-2013) under grant agreement 215009 RedCat. References [1] G. Pappa, H. Bartsch, and C. Gerhauser, Biphasic modulation of cell proliferation by sulforaphane at

physiologically relevant exposure times in a human colon cancer cell line, Mol. Nutr. Food Res. 51 (2007) pp. 977-984.

[2] N.E.B. Saidu, S. Valente, E. Bana, G. Kirsch, D. Bagrel, and M. Montenarh, Coumarin polysulfides inhibit cell growth and induce apoptosis in HCT116 colon cancer cells, Bioorg. Med. Chem. 20 (2012) pp. 1584-1593.

[3] M. Montenarh and N.E.B. Saidu, The effect of diallyl polysulfanes on cellular signalling cascades, Nat. Prod. Commun. 7 (2012) pp. 401-408.

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Diallyl tetrasulfane activates both the eIF2α and Nrf2/HO-1 pathways

N. E. B. Saidu, R. Touma, I. A. Asali, C. Jacob, M. Montenarh

Medical Biochemistry and Molecular Biology, University of the Saarland, Building 44, 66424 Homburg, Germany

Background: Diallyl polysulfanes have been shown to exert cell cycle arrest, anti-

tumour and anti-inflammatory activities in a variety of in vitro and in vivo models. Although diallyl polysulfanes cause oxidative stress, little is known about the underlying signaling cascades leading to antioxidant defence or apoptosis.

Methods: Cells were treated with DATTS at different concentrations and for different periods. Reactive oxygen species and thiol concentrations were determined by commercially available kits. The expression levels of signal molecules were determined by Western Blot analysis. A direct influence of Nrf2 on the promoter of HO-1 gene was determined by a luciferase assay with the StRE promoter element from the HO-1 gene.

Results: We found an immediate increase in the level of the superoxide anion radical O2

•- and hydrogen peroxide H2O2 and an overall thiol depletion. DATTS treatment of HCT116 cells also caused an up-regulation of phospho-eIF2α, nuclear Nrf2 and HO-1 protein levels in a time and concentration-dependent manner. Pre-treatment of cells with antioxidants significantly reduced the elevated expression levels of these proteins. A direct contribution of Nrf2 was shown by its interaction with the stress response element of the HO-1 promoter.

Conclusions: DATTS activates the ROS- eIF2α/Nrf2 HO-1 signaling cascades leading to the up-regulation of HO-1. However, this antioxidant defence is not sufficient to protect HCT116 cells from apoptosis.

General significance: This study shows for the first time a parallel but not equal activation of signaling pathways by DATTS with a competitive ultimate cellular outcome.

Keywords: reactive oxygen species (ROS), thiols, redox reagents, antioxidant pathway, apoptosis Acknowledgments: This work was supported by the European Community’s framework programme (FP7/2007-2013) under grant agreement 215009 RedCat. References [1] G. Pappa, H. Bartsch, and C. Gerhauser, Biphasic modulation of cell proliferation by sulforaphane at

physiologically relevant exposure times in a human colon cancer cell line, Mol. Nutr. Food Res. 51 (2007) pp. 977-984.

[2] N.E.B. Saidu, S. Valente, E. Bana, G. Kirsch, D. Bagrel, and M. Montenarh, Coumarin polysulfides inhibit cell growth and induce apoptosis in HCT116 colon cancer cells, Bioorg. Med. Chem. 20 (2012) pp. 1584-1593.

[3] M. Montenarh and N.E.B. Saidu, The effect of diallyl polysulfanes on cellular signalling cascades, Nat. Prod. Commun. 7 (2012) pp. 401-408.

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Diallyl polysulfides from garlic – mode of action and applications in agriculture

Miriam Arbach,a,b Awais Anwar,b Murree Groom,b Chris Hamiltona

aSchool of Pharmacy, University of East Anglia, Norwich NR4 7TJ, United Kingdom; bECOspray Limited,

Grange Farm, Hilborough, Norfolk IP26 5BT, United Kingdom

Garlic (Allium sativum) contains a wide range of organosulfur compounds which show a variety of biological effects including broad spectrum antibacterial, antifungal and antiviral activity, as well as selective anticancer activity.[1,2] When garlic is crushed, the enzyme alliinase converts alliin (S-allyl-L-cysteine sulfoxide) (1) into allicin (2), a highly reactive and therefore fairly unstable thiosulfoxide. Several degradation products are formed from allicin, e.g. cyclic dithiins (8-9), ajoenes (10) and diallyl polysulfides (3-7) (DAPS). Different biological activities are known for DAPS, but little is known about their mode(s) of action.

Here we report the results of studies investigating the influence of DAPS on the intracellular low molecular weight (LMW) thiol redox status. The work tests the theory of one possible mode of action of DAPS: When DAPS reach the cell, they react with thiols (e.g. LMW thiols or protein thiols), which leads on the one hand to a shift in the thiol redox status and on the other hand to the liberation of reactive hydropersulfides which can again react with thiols or lead to other follow on reactions in the cell (e.g. formation of reactive oxygen species).[3,4] To test this hypothesis a bacterial model organism Bacillus subtilis is used which produces bacillithiol (BSH) as major LMW thiol.[5] To gain an insight into the role of BSH in defence and/or activation of DAPS, studies have been performed with the wild type strain and a mutant strain that does not produce any BSH.

Additionally the activity of different polysulfide formulations towards plant pathogenic nematodes is investigated in bioassays as well as in potato and carrot field trials to exploit the polysulfide chemistry for the development of a “green” nematicide. References [1] International Journal of Oncology, 2010. 36(3): p. 743-749. [2] Organic & Biomolecular Chemistry, 2007. 5(10): p. 1505-1518. [3] Free Radical Biology and Medicine, 2003. 34(9): p. 1200-1211. [4] Physiologia Plantarum, 2008. 133(3): p. 469-480. [5] Nature Chemical Biology, 2009. 5(9): p. 625-627.

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The natural organosulfur dipropyltetrasulfide prevents systemic sclerosis in the mice

Wioleta Marut, Vincent Jamier, Niloufar Kavian, Amélie Servettaz, Paul Winyard, Paul

Eggleton, Awais Anwar, Carole Nicco, Christiane Chéreau, Bernard Weill, Frédéric Batteux

Université Paris Descartes, Faculté de Médecine, EA 1833 et Laboratoire d’immunologie biologique,

Hôpital Cochin AP-HP, 75679 Paris cedex 14, France

The pro-oxidative organosulfur molecule Dipropyltertrasulfide (DPTTS) is a natural compound found in Allium vegetable and devoted with antibiotic and anti-cancer properties.[1, 2] The aim of this study was to evalute the efficiency of DPTTS in a recently described mouse model of HOCl-induced SSc.

The pro-oxidative effects of DPTTS and their anti-proliferative and cytotoxic consequences were evaluated in vitro on mouse normal and SSc skin fibroblasts. The anti-fibrotic and immunomodulating properties of iv injection of DPTTS were evaluated in vivo in HOCl-induced SSc.

In vitro, the ROS production was higher in mouse SSc fibroblasts than in normal fibroblasts and was still increased by DPTTS to reach a cytostatic and cytotoxic threshold that kills mouse SSc fibroblasts.

SSc mice treated with DPTTS presented significant decrease in dermal thickness and collagen concentration in the skin compared to untreated SSc mice. Serum concentrations of advanced oxidation protein products, and hydogen sulfide were significantly reduced in SSc mice treated by organosulfur molecule. Moreover, in vivo treatment of HOCl mice with DPTTS reduced splenic B cell count, inhibited auto-antibodies production, decreased CD3/CD28 and LPS induced splenocytes proliferation and decreased the production of IL-4 and IL-13 produced by activated T cells.

We demonstrate that the natural organosulfur compound DPTTS prevents skin fibrosis in the mouse by the selective killing of diseased fibroblasts and its immunomodulating properties. Therefore, the natural vegetable-derived compound DPTTS could be a safe and effective treatment of SSc. Acknowledgments: This work was supported by European Community’s Seventh Framework Programme (FP7/2007-2013) under grant agreement 215009 RedCat for financial support. References [1] Groschel, B.; Bushman, F. Journal of virology 2005, 79(9):5695-5704. [2] Kelkel, M.; Cerella, C.; Mack, F.; Schneider, T.; Jacob C.; Schumacher, M.; Dicato, M.; Diederich, M.

Carcinogenesis 2012.

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New natural compounds of Dionaea muscipula as anticancer agents

François Gaascht,a Claudia Cerella,a Monika Jain,a Marie-Hélène Teiten,a Marc Schumacher,a Gilbert Kirsch,b Denyse Bagrel,c Mario Dicato,a Marc Diedericha,d

a Laboratoire de Biologie Moléculaire et Cellulaire du Cancer, Hôpital Kirchberg, 9 rue Edward Steichen, L-2540 Luxembourg, Grand Duchy of Luxembourg; b Laboratoire d’Ingénierie Moléculaire et Biochimie Pharmacologique, Université Paul Verlaine de Metz, Technopôle, 1 Boulevard Arago, F-57070, Metz,

France; c Laboratoire d’Ingénierie Moléculaire et Biochimie Pharmacologique, Université Paul Verlaine de Metz, Campus Bridoux, Rue du Général Delestraint, F-57070, Metz, France; d Department of Pharmacy,

College of Pharmacy, Seoul National University, Seoul, 151-742, KOREA

Since centuries, natural compounds are used in traditional medicine. Recent studies have shown that such substances can block appearance and development of many diseases including cancer. Because of emergence of new diseases and resistance towards conventional treatments, current drug library needs to be enlarged by discovery and characterization of new molecules.

The Venus Flytrap (Dionaea muscipula Solander Ex Ellis) is one of the best-known carnivorous plants that catches and digests small preys with an active trap. This plant has been shown to produce many therapeutic molecules that are also present in other plants (like myricetin, quercetin, plumbagin and ellagic acid) but to date no specific natural compound had been discovered in Dionaea muscipula. The aim of this project was to characterize specific therapeutic molecules from Dionaea muscipula and especially to study their apoptotic effects on leukemia and lymphoma cells.

Plumbagin or 5-hydroxy-2-methyl-1,4-naphthoquinone has been originally identified from Plumbago zeylanica, a plant used in traditional Indian, Chinese and African medicines and cultures. Today, plumbagin is known to exert antioxidant, anti-inflammatory, anti-tumor, antibacterial and antifungal activities properties and to modulate several cell signaling pathways including pathways modulating cell death by apoptosis. Nevertheless, mechanisms leading to apoptosis remain unclear. In this context, we have chosen to investigate in details the effect of plumbagin on the apoptotic pathway.

We have focused our work on a plumbagin-sensitive cancer cell line, U937 and on a plumbagin-resistant cancer cell line, Raji. Our results have shown that plumbagin induced apoptosis in both cell lines, with a concentration of respectively 1 µM in U937 cells and 4 µM in Raji cells.

A stringent time-course analysis in U937 cells revealed a specific pattern of modulation of the anti-apoptotic Bcl-2 family proteins with Bcl-2 proteolysis and Mcl-1 down-regulation which take place concomitantly with the triggering of major caspases cleavage and massive apoptosis occurrence. The same pattern of modulation might be observed in Raji cells when treated with apoptogenic concentrations of plumbagin. Further studies are in course to elucidate the causative or amplifying roles of these modulations in plumbagin-induced apoptotic signaling.

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Exploring the versatility of (E,E)-cinnamylideneacetophenones as templates for the synthesis of potentially bioactive compounds

Diana I. S. P. Resende, Cristina G. Oliva, Artur M. S. Silva, José A. S. Cavaleiro

Department of Chemistry & QOPNA, University of Aveiro, 3810-193 Aveiro, Portugal

(E,E)-cinnamylideneacetophenones 1 are a well-known group of unsaturated ketones

that have been widely used as templates in several synthetic transformations,[1] and proved to be versatile precursors of potentially bioactive compounds.[1b,2] In the last years, our group become interested in the 1,4-conjugation addition of different nucleophiles to these α,β,γ,δ-unsaturated ketones 1 and also to study their behavior domino multicomponent reactions and under phase transfer catalysis.

In this communication we describe our results on the 1,4-conjugate addition of nitromethane and malononitrile to (E,E)-cinnamylideneacetophenones 1. We were able to control the reaction enantioselectivity with different organocatalysts and additives obtaining the building blocks 2 and 3 in excellent yields and enantioselectivities. These compounds 2 and 3 can be converted into therapeutic interesting compounds, such as polysubstituted cyclohexane derivatives 4 and 5, via a Michael/aldol reaction with the α,β,γ,δ-unsaturated ketones 1.[3,4] Interestingly we found that these derivatives 4 and 5 can also be generated directly from the (E,E)-cinnamylideneacetophenones 1 via a Michael/Michael/addition with the appropriate nucleophile.

Acknowledgments: Thanks are due to the University of Aveiro, FCT and FEDER for funding the Organic Chemistry Research Unit. (project PEst-C/QUI/UI0062/2011) and the Portuguese National NMR Network (RNRMN). D. I. S. P. Resende thanks also FCT for her PhD grant (SFRH/BD/62696/2009). References [1] (a) Santos, C. M. M.; Silva, A. M. S.; Cavaleiro, J. A. S.; Lévai, A.; Patonay. T. Eur. J. Org. Chem.

2007, 17, 2877. (b) Pinto, D.C.G.A.; Silva, A.M.S.; Lévai, A.; Cavaleiro, J.A.S.; Patonay, T.; Elguero. J. Eur. J. Org. Chem. 2000, 14, 2593.

[2] Silva, A. M. S.; Pinto, D. C. G. A.; Tavares, H. R.; Cavaleiro, J. A. S.; Jimeno, M. L.; Elguero, J. Eur. J. Org. Chem. 1998, 2031.

[3] (a) Oliva, C. G..; Silva, A. M. S.; Paz, F. A. A.; Cavaleiro, J. A. S. Synlett 2010, 1123. (b) Oliva, C. G..; Silva, A. M. S.; Resende, D. I. S. P.; Paz, F. A. A.; Cavaleiro, J. A. S. Eur. J. Org. Chem. 2010, 18, 3449.

[4] (a) Resende, D. I. S. P.; Oliva, C. G..; Silva, A. M. S.; Paz, F. A. A.; Cavaleiro, J. A. S. Synlett 2010, 115. (b) Oliva, C. G..; Resende, D. I. S. P.; Silva, A. M. S.; Cavaleiro, J. A. S., in preparation.

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4-Vinyl-2H-1-benzopyran-2-one reactivity as [4+2] thermal Diels-Alder cycloaddition: access to novel coumarin-based polycycles with Cdc25

phosphatases inhibiting activity

Sergio Valente,a,b,c Zhanjie Xu,d,e Emilie Bana,b Clemens Zwergel,b Antonello Mai,a Claus Jacob,d Peter Meiser,e Denise Bagrel,b Artur M. S. Silva,c Gilbert Kirschb

aDipartimento di Chimica e Tecnologie del Farmaco, Sapienza Università di Roma, P.le A. Moro 5, 00185

Roma; bLaboratoire d’Ingénierie Moléculaire et Biochimie Pharmacologique, Institut Jean Barriol, Université de Lorraine, 1 Boulevard Arago, 57070 Metz, France ; cDepartment of Chemistry & QOPNA,

University of Aveiro, 3810-193 Aveiro, Portugal; dDivision of Bioorganic Chemistry, School of Pharmacy, Saarland University, D-66123 Saarbrücken, Germany; eUrsapharm Arzneimittel GmbH & Co KG,

Industriestraße 35, 66129 Saarbrücken Germany

So far a large number of natural products bear a 2H-1-benzopyran-2-one (coumarin) as part of their structure.[1] 2H-1-benzopyran-2-ones show wide and well reviewed biological activity.[2] Recently our group has optimized the method to provide methyl ketone at the C4 position on the 2H-1-benzopyran-2-one scaffold through very high α-regioselective Heck coupling reaction using tosylates as substrates.[3] As an intermediate we obtained 4-(1-butoxyvinyl)-2H-chromen-2-one 1, a very useful diene. Otherwise, when we performed the cross-coupling on 4-bromo-coumarin, the reaction regioselectively afforded the beta isomer (E)-4-(2-butoxyvinyl)-2H-chromen-2-one 2, and not a mixture of both isomer, as expected. Hence, we studied dienes 1 and 2 reactivity in (4π+2π) thermal Diels-Alder cycloaddition reaction with several dienophiles aiming to build novel coumarin-containing polycycles. As in 2010 we reported some coumarin-based derivatives as Cdc25 phosphatases inhibitors,[4] we decided to test some of the new polycycles against the three Cdc25 isoforms, respectively named A, B and C. Cdc25 phosphatases are key enzymes regulating the cell cycle and represent valuable target for cancer treatment. The biological activity of such compounds against these enzymes and in cancer cell lines will be presented.

References [1] Kostova, I. Curr. Med. Chem. Anticancer Agents 2005, 5, 29-46. [2] Riveiro, ME et al. Curr Med Chem. 2010;17, 1325-38. [3] Valente, S. and Kirsch, G. Tetrahedron Lett. 2011, 52, 3429-3432. [4] Valente, S. Et al. Bioorg. Med. Chem. Lett. 2010, 20, 5827-30.

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Human peroxiredoxin and oxidative stress

Paul James, Misha Isupov, Jennifer Littlechild

Henry Wellcome Building for Biocatalysis, Biosciences, College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD,UK.

Peroxiredoxins (Prx) are a family of multifunctional enzymes that protect against oxidative stress and are produced in high levels in cells. Prx form part of a peroxide scavenging system along with sulfiredoxin (Srx), thioredoxin (Trx) and thioredoxin reductase (TrxR). All these enzymes use redox active cysteine residues to detoxify reactive oxygen species or reduce the other enzymes in the system. The structure of the red blood cell hPrxII was solved in Exeter and shows a decameric structure with the peroxidatic cysteine residue oxidised to sulfinic acid.[1] As part of the mechanism of this enzyme the peroxidatic and the resolving cysteine from its neighbouring subunit form an intermolecular disulfide bond. In the crystal structure of hPrxII the peroxidatic cysteine is found in the hyper-oxidised sulfinic acid form. In this form the cysteine residues are ~10 Å apart requiring a large structural movement to allow them to be close enough for the bond to form.

The original hPrxII protein was isolated directly from human blood. The hPrxII protein has now been over-expressed in Escherichia coli[2] and purified in the presence of the oxidising agent diamide. The protein was still found to be in the decameric form (using a calibrated gel filtration column). The recombinant protein was crystallized and the structure solved to 3.3 Å resolution which has allowed the necessary structural rearrangements to be observed.[2,3] This new structure shows the decameric form of hPrxII in the disulfide state that is known to form as part of its active cycle. It has identified movements that have to take place to bring the peroxidatic and resolving cysteine residues together (previously ~13Å apart) to form the disulfide bond. It was previously believed that the decameric form of the protein could not exist in the disulfide state and only formed when the Prx was a dimer.

To understand the interaction between the enzymes involved in this peroxide scavenging system, covalent complexes were constructed between hPrxII and Trx and hPrxII and Srx. All of the cysteine residues in these proteins were removed by site-directed mutagenesis except for those involved in disulfide formation. The proteins were encouraged to form a covalent bond by first reacting peroxiredoxin with 5,5’-dithiobis-(2-nitrobenzoic acid) (DTNB) and then replacing TNB with the protein of interest. The complex molecular weight was estimated using a calibrated gel filtration column. The size of the complex between the decameric form of PrxII and Trx was ~370 kDa. The PrxII has a molecular weight of ~250 kDa leaving ~120 kDa. This would equate to 10 Trx monomers which would decorate each subunit of the PrxII in the decamer. The size of the complex between the decameric form of PrxII and Srx is ~315 kDa. The PrxII has a weight of ~250kDa leaving ~65 kDa of Srx which would equate to 5 Srx monomers allowing one Srx to interact with each PrxII dimer. These studies have provided new information on the stoichiometry of the transitional binding between the protein components of this hPrxII peroxide scavenging system.[4] References [1] E. Schroder, J. Littlechild, A. Lebedev, N. Errington, A. Vagin and M. Isupov. Structure (2000) 8, 605-

615. [2] P. James, PhD thesis University of Exeter, 2011. [3] P. James, K. Hamidi, C. Rye, K.Line and J.Littlechild (2012) Protein Express. Purif. Submitted. [4] P. James, K. Line, M. Isupov and J. Littlechild (2012) Manuscript in preparation.

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Synthesis of flavone derivatives with potential biological activity

Djenisa H. A. Rocha, Diana C. G. A. Pinto, Artur M. S. Silva, José A. S. Cavaleiro

Department of Chemistry & QOPNA, University of Aveiro, 3810-193 Aveiro, Portugal

Plants are the main source of numerous important compounds; their importance is mainly due to its diverse biological properties. We can draw attention to flavones, natural compounds belonging to the flavonoid family[1] and presenting pharmacological properties, such as antibacterial, anti-inflammatory, anticancer, antiviral and antioxidant activities[1,2] which have led to the development of new synthetic approaches for their synthesis. Their antioxidant potential has been exhaustly studied and it has been established that the presence of hydroxyl groups at certain position on the skeleton, for instance a B ring catechol moiety and a 3- and 5-hydroxyl substituents, and also a C2=C3 double bond conjugated with the 4-carbonyl group are important structural features for scavenging superoxide anion, hydroxyl radical and peroxyl radical or quenching singlet oxygen and inhibiting lipid peroxidation in the biological system in vitro.[3]

Having in mind the referred structure-antioxidant activity relationship and considering that we have been dedicate our investigation to find out new antioxidant derivatives we have set up a program to synthesise flavone derivatives that would present the ideal type of substituent’s and can be regarded as future new antioxidant compounds and/or intermediate of other important ones.[4a] The methods that we are developing and that will be presented and discussed in this communication involve the Wittig and Heck reaction to obtain 3-styrylflavones 1 and the Sonogashira reaction to obtain 3-arylethynyl-flavones 2.[4]

Acknowledgments: Thanks are due to the University of Aveiro, FCT and FEDER for funding the Organic Chemistry Research Unit. D. H. A. Rocha (SFRH/BD/68991/2010) also thanks FCT for their PhD grant. References [1] Martens, S.; Mithöfer, A. Phytochemistry 2005, 66, 2399 and references cited there in. [2] (a) Pan, M.-H.; Lai, C.-S.; Ho, C.-T. Food Funct. 2010, 1, 15. (b) Kamal, A.; Murty, J. N. S. R.;

Viswanath, A.; Sujitha, P.; Kumar, C. G. Bioorg. Med. Chem. Lett., 2012, 22, 4891. [3] (a) Yi, L.; Chen, C.y.; Jin, X.; Zhang, T.; Zhou, Y.; Zhang, Q.-y.; Zhu, J.-d.; Mi, M.-t.; Biochimie 2012,

94, 2035. (b) Marković, J. M. D.; Marković, Z. S.; Pašti, I. A.; Brdarić, T. P.; Popović-Bijelić, A.; Mojović, M.; Dalton Trans. 2012, 41, 7295.

[4] (a) Rocha, D. H. A.; Pinto, D. C. G. A.; Silva, A. M. S.; Patonay, T.; Cavaleiro, J. A. S.; Synlett, 2012, 23, 559. (b) Seixas, R. S. G. R.; Pinto, D. C. G. A.; Silva, A. M. S.; Cavaleiro, J. A. S.; Aust. J. Chem.; 2008, 61, 718. (c) Sandulache, A.; Silva, A. M. S.; Pinto, D. C. G. A.; Almeida, L. M. P. M.; Cavaleiro, J. A. S.; New J. Chem. 2003, 27, 1592.

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Studies on the chlorination of flavones: synthesis of chlorohydroxyflavones with potential biological activity

Sara M. Tomé, Artur M. S. Silva

Department of Chemistry & QOPNA, University of Aveiro, 3810-193 Aveiro, Portugal

Flavones (2-phenylchromones) are one of the major classes of natural heterocyclic

compounds. They are known to exhibit a wide range of biological properties, including anti-oxidant, anti-inflammatory, cancer suppressing and antiviral (anti-HIV) activities,[1] and represent a strong backbone for the synthesis of pharmacological active molecules. On the other hand, it is known that the halogenation of an active molecule may enhance its biological activity.[2] The truth is that many pharmaceutical drugs[3] and agrochemicals[4] are halo-substituted compounds.

In the present communication we present and discuss the experimental attempts towards the synthesis of flavones 4a-c. Chlorination of flavone 2 (obtained by the oxidative cyclization of 2’-hydroxychalcone 1) was performed by two different methods depending on the aimed chlorination pattern (Scheme 1). Flavones 3a,b were successfully synthesized by using N-chlorosuccinimide as the chlorination agent and the synthesis of flavone 3c was accomplished by using NaOCl (10%). The final step consists on the cleavage of the methyl protecting groups of flavones 3a-c with boron tribromide affording the expected chlorohydroxyflavones 4a-c, which are currently under biological evaluation.

Scheme 1 Acknowledgments: Thanks are due to the University of Aveiro, FCT and FEDER for funding the Organic Chemistry Research Unit (project PEst-C/QUI/UI0062/2011). Sara Tomé also thanks University of Aveiro and Organic Chemistry Research Unit for her fellowship (BIIC/QUI/5173/2011). References [1] Rice-Evans, C. A.; Packer, L. Flavonoids in Health and Disease, 2nd ed., Marcel Dekker, New York,

2003. [2] (a) Binsack, R.; Boersma, B. J.; Patel, R. P.; Kirk, M.; White, C. R.; Darley-Usmar, V.; Barnes, S.; Zhou,

F.; Parks, D. A. Alcohol Clin. Exp. Res. 2001, 25, 434. (b) Marder, M.; Viola, H.; Wasowski, C.; Wolfman, C.; Waterman, P. G.; Cassels, B. K.; Medina, J. H.; Paladini, A. C. Biochem. Biophys. Res. Commun. 1996, 223, 384. (c) Park, H.; Dao, T. T.; Kim, H. P. Eur. J. Med. Chem. 2005, 40, 943.

[3] Abraham, D. J. Burger’s Medicinal Chemistry and Drug Discovery, Vol. 5, 6th ed., Wiley, New Jersey, 2003.

[4] Baker, D. R.; Basarab, G. S.; Fenyes, J. G. Synthesis and chemistry of agrochemicals IV, ACS Symposium Series 584, American Chemical Society, Washington, 1995.

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Azines derived from C-formyl-1H-imidazoles: synthesis and structural studies in solution and solid state NMR

Joana Pinto,a Vera L. M. Silva,a Artur M. S. Silva,a Rosa M. Claramunt,b Dionisia Sanz,b M. Carmen Torralba,c M. Rosario Torres,c Felipe Reviriego,d Ibon Alkorta,d José Elguerod

aDepartment of Chemistry & QOPNA, University of Aveiro, 3810-193 Aveiro, Portugal; bDepartamento de Química Orgánica y Bio-Orgánica, Facultad de Ciencias, Universidad Nacional de Educación a Distancia

(UNED), Senda del Rey 9, E-28040 Madrid, Spain; cDepartamento de Química Inorgánica I and CAI de Difracción de Rayos-X, Facultad de Ciencias Químicas, Universidad Complutense de Madrid (UCM), E-

28040 Madrid, Spain; dInstituto de Química Médica, Centro de Química Orgánica "Manuel Lora-Tamayo", CSIC, Juan de la Cierva, 3, E-28006 Madrid, Spain

Azines, the compounds resulting from the double condensation of carbonyl

compounds with hydrazine (aldazines and ketazines derived from aldehydes and ketones, respectively) are much studied compounds with special emphasis on their use as synthons, [1] on their mesogenic (as such or with metals, metallomesogens) and NLO properties,[2] as well as for their interesting structural properties.[3] Some authors have reported biological properties of azines,[4] but the most important azine in what concerns activity is hydramethylnon a powerful insecticide that owing to its azine bridge decomposes rapidly in water.[5]

Here we present our results on the synthesis of three aldazines 1-3 derived from C-formylimidazoles, being one of them doubly labeled with 15N (2L). The results of structural studies in solution and solid state NMR will also be reported and discussed.

N

N N

H

N

H

N

N

H

H

N

N N

H

N

H

N

N

H3C

CH3

H

H

N

N N

H

N

H

N

N

H

H

N

N N

H

N

H

N

N

H

H 15

15

1 2 2L 3

12

34

5 6

7

7'

6'2'

1'

3'4'

5'1

2

34

56

7

7' 7'

7

65

43

2 1

1' 1'2' 2'

3' 3'4' 4'

5' 5'6' 6'

Acknowledgments: Thanks are due to the University of Aveiro, ‘Fundação para a Ciência e Tecnologia’ and FEDER for funding the Organic Chemistry Research Unit (project PEst-C/QUI/UI0062/2011) and the Portuguese National NMR Network (RNRMN). This work has also been financed by the Spanish MICINN (CTQ2009-13129-C02-02 and CTQ2010-16122) and by the Comunidad Autónoma de Madrid (Project MADRISOLAR2, ref S2009/PPQ-1533). Joana Pinto (SFRH/BD/77807/2011) and Vera Silva (SFRH/BPD/27098/2006) thanks FCT for their grants. References [1] (a) Cohen, R.; Rybtchinski, B.; Gandelman, M.; Shimon, L. J. W.; Martin, J. M. L.; Milstein, D. Angew.

Chem. Int. Ed. 2003, 42, 1949. (b) Deun, R. V.; Parac-Vogt, T. N.; Van Hecke, K.; Van Meervelt, L. V.; Binnemans, K.; Guillon, D.; Donnio, B. J. Mater. Chem. 2003, 13, 1639. (c) Burger, K.; Hennig, L.; Zeika, O.; Lux, A. Heterocycles 2006, 67, 443.

[2] Moreno-Mañas, M.; Pleixats, R.; Andreu, R.; Garín, J.; Orduña, J.; Villacampa, B.; Levillan, E.; Sallé, M. J. Mater. Chem. 2001, 11, 374.

[3] Wolstenholme, D. J.; Cameron, T. S. J. Phys. Chem. (A) 2006, 110, 8970. [4] Kurteva, V. B.; Simeonov, S. P.; Stoilova-Disheva, M. Pharmacol. Pharm. 2011, 2, 1. [5] Unger, T. A. Pesticide Synthesis Handbook, Noyes Publications, New Jersey, 1996, p. 585.

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New approaches to the synthesis of benzo[b]acridin-12(7H)-ones and 3-styrylquinolin-4(1H)-ones

Raquel S. G. R. Seixas, Artur M. S. Silva, Diana C. G. A. Pinto, José A. S. Cavaleiro

Department of Chemistry & QOPNA, University of Aveiro, 3810-193 Aveiro, Portugal

raquel.grevy@ua.pt

Quinolin-4(1H)-ones and acridones are important families of naturally occurring nitrogen heterocyclic compounds exhibiting a variety of important biological activities. Quinolin-4(1H)-ones have been the subject of extensive study due to their potential biological applications,[1] but their use is mainly as broad-spectrum antibiotics.[2] Acridone derivatives are known to present important antiviral, antiparasitic and anticancer activities. [3] The variety of important potential biological applications of quinolin-4(1H)-ones and acridones and the continuous search of the scientific community for the development of new molecules with attractive biological activities highlight these compounds as targets for the preparation of new derivatives or/and to develop new strategies for their synthesis.

In this communication we will describe the synthesis of 3-styrylquinolin-4(1H)-ones and benzo[b]acridin-12(7H)-ones using 4-chloroquinoline-3-carbaldehyde 1 as starting material. The (Z)-3-styrylquinolin-4(1H)-ones 3 are obtained with high diastereoselectivity from the Wittig reaction of N-protected-quinolin-4(1H)-one-3-carbaldehydes 2 while (E)-3-styrylquinolin-4(1H)-ones 4 are prepared through the Wittig reaction of 4-chloroquinoline-3-carbaldehyde 1 followed by acid hydrolysis.[4] Benzo[b]acridin-12(7H)-ones 5 are obtained through the Diels-Alder reaction of ortho-benzoquinodimethanes with N-protected-quinolin-4(1H)-one-3-carbaldehyde 2, followed by oxidation.[5] Experimental procedures and structural elucidation of synthesised compounds will be presented and discussed in this communication.

Acknowledgments: Thanks are due to the University of Aveiro, FEDER and FCT for funding the project POCI/QUI/58835/2004, the Organic Chemistry Research Unit and the Portuguese National NMR Network (RNRMN). Raquel S. G. R. Seixas also thanks FCT for a PhD (SFRH/BD/30734/2006) grant. References [1] (a) Hsu, S.C. et al. J. Orthop. Res. 2009, 27, 1637; (b) Sato, M. et al. J. Med. Chem. 2009 52, 4869; (c)

Pasquini, S. et al. J. Med. Chem. 2008, 51, 5075. [2] Alós, J.-I. Enferm. Infecc. Microbiol. Clin. 2003, 21, 261. [3] (a) Winter, R. W. et al. Exp. Parasitol. 2006, 114, 47; (b) Itoigawa, M. et al. Cancer Lett. 2003, 193, 133. [4] Seixas, R.S.G.R., Silva, A.M.S., Cavaleiro, J.A.S. Synlett 2010 2257. [5] Seixas, R.S.G.R., Silva, A.M.S., Pinto, D.C.G., Cavaleiro, J.A.S. Synlett 2008 3193.

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Synthetic strategies towards novel acridin-9(10H)-ones and (E)-2-aryl-4-styrylfuro[3,2- c]quinolines

Vera L. M. Silva, Artur M. S. Silva, José A. S. Cavaleiro

Department of Chemistry & QOPNA, University of Aveiro, 3810-193 Aveiro, Portugal

Acridin-9(10H)-ones and furoquinolines are characteristic secondary metabolites of

plants of the Rutaceae family.[1] Several naturally occurring and synthetic derivatives of acridin-9(10H)-ones are known due to their biomedical potential including antitumoral,

antioxidant, anticancer, antimicrobial, antiviral, mutagenic and cytotoxic activities.[2] Quinoline derivatives, including furoquinolines, have shown in vitro activity against cutaneous and visceral leishmaniasis, African trypanosomiasis and Chagas disease, among other important biological activities.[3]

Several methods for the synthesis of linear furo[2,3-b]quinolines have been reported, but in contrast the synthesis of the isomeric angular furo[3,2-c]quinolines have been much less investigated. On the other hand, most of the synthetic approaches commonly used for the synthesis of acridin-9(10H)-ones are regioselectivity-compromised and usually require harsh acid mediated conditions.

Here we present our recent achievements in the development of new synthetic routes to prepare novel 2,3-diarylacridin-9(10H)-ones 1, 2, 1-arylacridin-9(10H)-ones 3 and (E)-2-aryl-4-styrylfuro[3,2-c]quinolines 4.[4] When properly functionalized these compounds have a great potential as antioxidant, antitumor and antiviral agents.

N

O

R1

1: R1 = H

2: R1= CH3

R2 = H, OCH3, Cl

R3, R4 = H, OCH3

R2

R3

R4

N

O

CH3

R

3: R = H, OCH3, NO2, Cl

N

O

R3

R2

R1

4: R1 = H, OCH3, Cl

R2, R3 = H, OCH3 Acknowledgments: Thanks are due to the University of Aveiro, “Fundação para a Ciência e a Tecnologia” (FCT) and FEDER for funding the Organic Chemistry Research Unit (project PEst-C/QUI/UI0062/2011) and to the Portuguese National NMR network also funded by FCT. Vera L. M. Silva also thanks FCT for the grant (SFRH/ BPD/ 27098/ 2006). References [1] Kamdem Waffo, A. F.; Coombes, P. H.; Crouch, N. R.; Mulholland, D. A.; El Amin, S. M. M.; Smith, P.

J. Phytochemistry 2006, 68 (5), 663. [2] (a) Michael, J. P.; Nat. Prod. Rep. 2007, 24, 223; (b) Wansi, J. D.; Wandji, J.; Mbaze Meva, L.; Kamdem

Waffo, A. F.; Ranjit, R.; Khan, S. N.; Asma, A.; Iqbal, C. M.; Lallemand, M.-C.; Tillequin, F.; Tanee Z. F. Chem. Pharm. Bull., 2006, 54 (3), 292. (c) Nguyen, H. T.; Lallemand, M.-C.; Boutefnouchet, S.; Michel, S.; Tillequin, F. J. Nat. Prod. 2009, 72, 527.

[3] (a) Suarez, L. E. C.; Pattarroyo, M. E.; Lozano, J. L.; Monache. F. D. Nat. Prod. Res. 2009, 23 (4), 370. (b) Hanawa, F.; Fokialakis, N.; Skaltsounis, A. L. Planta Med., 2004, 70 (6), 531.

[4] Silva, V. L. M.; Silva, A. M. S.; Cavaleiro, J. A. S. Synlett, 2010, 17, 2565.

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Visible light-induced (E Z)-photoisomerization of benzofuran-3-one-hydantoin hybrids

Guido R. Lopes, Oualid Talhi, Diana C. G. A. Pinto, Artur M. S. Silva

Department of Chemistry & QOPNA, University of Aveiro, 3810-193 Aveiro, Portugal

The (E�Z)-isomerization are important issues in chemistry and biology fields

offering various industrial, technological and medicinal applications.[1] The (E�Z)-isomerism phenomenon is usually induced by photo- and/or thermal energy contribution or even chemically catalysed.[2] Regarding to this research area, we describe the diastereoselective photo-dependent (E�Z) and (Z�E) isomerization equilibrium of (E/Z)-1,3-ditolyl-5-(3-oxobenzofuran-2(3H)-ylidene)imidazolidine-2,4-dione (DTBI). The photochemical process, which occurs spontaneously in solution when exposed to direct visible-light (sunlight or classic electrical lamp), was investigated by 1H NMR spectroscopy and HPLC-UV spectrophotometry. Both diastereomers (E)-DTBI and (Z)-DTBI are stereochemically stable in solid state, but show significant photo-sensibility in organic solutions. 1H NMR analysis of CDCl3 solutions of pure (E)-DTBI and (Z)-DTBI in the dark displays remarkable differences in their proton chemical shift values. Therefore, the photoisomerization equilibrium of DTBI can simply be deduced from the 1H NMR profile after visible-light irradiation of NMR tube solutions of both (E)-DTBI and (Z)-DTBI, indicating their presence in a 1:3 ratio. The photoisomerization reaction was monitored by HPLC-UV underling various influencing parameters such as light source, concentration, temperature and solvent effects. The results of the kinetic study reveals that (E Z)-equilibrium is mainly photo-dependent being the transformation (E�Z) proceeding faster than the (Z�E) in such circumstances. The (E Z)-equilibrium ratio depends on the used solvent, light source and temperature. Thermal heating does not induce photochemical transformation in the absence of light; moreover, UV irradiation has no effects as well. Thus, we demonstrated herein that both diastereomers are visible-light photo-sensitive tending to co-exist together in equilibrium solutions at a determined ratio which minimize the thermodynamic stability and increase the entropy of the system. However, in all the studied conditions, the equilibrium was always shifting to the most stable (Z)-product.

Acknowledgments: Thanks are due to the University of Aveiro, FCT and FEDER for funding the Organic Chemistry Research Unit (project PEst-C/QUI/UI0062/2011) and the Portuguese National NMR Network (RNRMN). The authors also thank the (European Community’s) Seventh Framework Programme (FP7/2007-20139 under grant agreement nº 215009) for the financial support. References [1] (a) Wald, G. Science, 1968, 162, 230; (b) Oesterhelt, D.; Stoeckenius, W. Nature, 1971, 233, 149; (c)

Oesterhelt, D.; Stoeckenius, W. Proc. Natl. Acad. Sci., 1973, 70, 2835; (d) Braun, A. M., Maruette, M. T.; Oleveros, E. Photochemical technology, New York: Wiley, 1991, 12, pp. 500; (e) Kirk-Othmer Encyclopedia of chemical technology, 4th edition, New York: Wiley, 1996, 18, pp. 799; (f) Ulmann Encyclopedia of chemical technology 5th edition, New York: VCH, 1991, A19, pp. 573.

[2] Inoue, Y.; Mori, T. C=C Photoinduced Isomerization Reactions, Synthetic Organic Photochemistry, Ed. J. Mattay, A. G. Griesbeck, CRC Press, 2004, p. 417–452.

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Synthesis and sensor applications of corroles with medium size carbon chains

Joana F. B. Barata,a Carla I. M. Santos,a,b M. Amparo F. Faustino,a Carlos Lodeiro,b M.

Graça P. M. S. Neves,a José A. S. Cavaleiroa

a Department of Chemistry and QOPNA, University of Aveiro, Campus de Santiago, 3810-193 Aveiro Portugal; bBIOSCOPE Group, Faculty of Science, Physical-Chemistry Department, Ourense Campus,

University of Vigo, 32004, Ourense, Spain.

Corroles are tetrapyrrolic macrocycles that share close similarities with porphyrins and other related macrocycles. The synthesis of new corrole molecules is always an exciting topic, due to its unpredictable chemistry, which is different from the porphyrin chemistry. Although corroles have been investigated in several areas[1] their ability as sensors have recently been disclosed.[2] Metal recognition plays an important role in supramolecular chemistry with their quantification really being important in the chemical industry, environmental science and biochemistry. Namely, mercury ion (Hg2+) is one of the important inorganic cations which have had a great effect on both environment and human body.

Following our interest on corrole derivatization,[3] we will report here the peculiar reaction of 5,10,15-tris(pentafluorophenyl)corrole with sarcosine and paraformaldehyde in different alcohols, affording new corrole derivatives bearing ether groups. The ability of these compounds as mercury ion sensors was also evaluated.

The experimental procedures, the structural characterization of the new compounds and the mechanistic considerations will be discussed. Acknowledgments: Thanks are due to the University of Aveiro, “Fundação para a Ciência e a Tecnologia” (FCT) and POCI 2010 (FEDER) for funding the Organic Chemistry Research Unit (Project PEst-C/QUI/UI0062/2011). J. F. B. Barata and C. I. M. Santos thank FCT for respectively their Post-Doctoral and doctoral grants SFRH/BPD/63237/2009 and SFRH/BD/64155/2009. References [1] Harel, I. A;. . Gross, Z; Chem. Eur. J. 2009, 15, 8382. [2] Santos, C. I. M.; Oliveira, E.; Barata, J. F. B., Faustino, M. A. F.; Cavaleiro, J. A. S.; Neves, M. G. P. M.

S.; Lodeiro, C. J. Mater. Chem. 2012, 22, 13811. [3] Barata, J. F. B; Neves, M. G. P. M. S.; Tomé, A. C.; Cavaleiro, J. A. S.; J. Porphyrins Phthalocyanines

2009, 13, 415.

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Studies on the influence of tetrapyrrolic nitrones in 1,3-dipolar cycloaddition reactions

A. Filipa F. Silva,a Joana F. B. Barata,a Sérgio M. Santos,b Ana M. G. Silva,c M. Graça P.

M. S. Neves,a Augusto C. Tomé,a Artur M. S. Silva,a José A. S. Cavaleiroa

aDepartment of Chemistry & QOPNA, University of Aveiro, 3810-193 Aveiro, Portugal; bDepartment of Chemistry & CICECO, University of Aveiro, 3810-193 Aveiro, Portugal; cREQUIMTE, Departamento de

Química e Bioquímica, Faculdade de Ciências, Universidade do Porto, 4169-007 Porto, Portugal

Studies on novel tetrapyrrolic molecules are being carried out in several areas, from chemistry to medicine, catalysis, sensors.[1] Knowing that their unique features are strongly dependent on their structures, new promising compounds with therapy applications can be synthesized by peripheral functionalization of the macrocycles.[2]

Following our interest in studying the behavior of porphyrins and corroles in 1,3-dipolar cycloaddition reactions,[2] we decided to synthesize tetrapyrrolic nitrones, through the reaction of 5,10,15,20-tetraphenylporphyrin-2-carbaldehyde and gallium(III)(pyridine) complex of 5,10,15-tris(pentafluorophenyl)corrole-3-carbaldehyde with N-methylhydroxyl--amine hydrochloride and to evaluate their reactivity features with dimethyl acetylenedicarboxylate.

Experimental procedures, mechanistic considerations with theoretical studies and spectroscopic data of the new compounds will be presented and discussed.

Acknowledgments: Thanks are due to the University of Aveiro, to Fundação para a Ciência e a Tecnologia (FCT) and FEDER for funding the Organic Chemistry Research Unit (QOPNA). J. F. B. B. also thanks FCT for her postdoctoral grant SFRH/BPD/63237/2009 References [1] (a) Kadish, K. M.; Smith K. M.; Guilard, R.; The Porphyrin Handbook-Applications: Past, Present and

Future, vol. 6. Academic Press: New York, 2000. (b) Harel, I. A;. . Gross, Z; Chem. Eur. J. 2009, 15, 8382.

[2] (a) Barata, J. F. B; Neves, M. G. P. M. S.; Tomé, A. C.; Cavaleiro, J. A. S.; J. Porphyrins Phthalocyanines 2009, 13, 415; (b) Silva, A. M. G.; Tomé, A. C.; Neves, M. G. P. M. S.; Silva, A. M. S.; Cavaleiro, J. A. S.; J. Org. Chem., 2005, 70, 2306. b) L. S. H. P. Vale, J. F. B. Barata, C. I. M. Santos, M. G. P. M. S. Neves, Faustino, M. A. F.; Tomé, A. C.; Silva, A. M. S.; Paz, F. A. A.; Cavaleiro, J. A. S.; J. Porphyrins Phthalocyanines 2009, 13, 358. c) Cavaleiro, J. A. S.; Tomé, A. C.; Neves, M. G. P. M. S.; Handbook of Porphyrin Science (With Applications to Chemistry, Physics, Materials Science, Engineering, Biology and Medicine), K. M. Kadish, K. M. Smith, R. Guilard, Eds., vol. 2, p. 193, World Scientific Publishing Co., Singapore, 2010.

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Porphyrin and phthalocyanine galacto-dendritic conjugates: powerful generators of reactive oxygen species to kill cancer cells

Patrícia M. R. Pereira,a,b Sandrina Silva,a José A. S. Cavaleiro,a Carlos A. F. Ribeiro,b João

P. C. Tomé,a Rosa Fernandesb

aDepartment of Chemistry & QOPNA, University of Aveiro, Portugal; bIBILI, Pharmacology & Experimental Therapeutics, Faculty of Medicine, University of Coimbra, Portugal

Porphyrins (Pors) and phthalocyanines (Pcs) are well known first and second

generation photosensitizers (PSs), respectively.[1] The combination of these PSs with light at a specific wavelength and molecular oxygen results in the formation of reactive oxygen species (ROS).[2] The efficacy of these PSs to target cancer cells and to generate ROS inside them, namely singlet oxygen (1O2) is crucial in cancer treatment by photodynamic therapy (PDT).[3] Our research group recently reported the synthesis of two powerful PSs: a porphyrin (PorGal8) and a phthalocyanine (PcGal16) decorated with eight and sixteen galactose units, respectively.[4]

In this communication, the ability of PorGal8 and PcGal16 to generate 1O2 will be reported, as well as their photo-activity against the human bladder cancer cell lines UM-UC-3 and HT-1376. Both PSs evidenced to be strong generators of 1O2 and photo-toxic to UM-UC-3 and HT-1376 cells after light irradiation. After PDT, it was also observed an increase in intracellular ROS production. The addition of specific quenchers of 1O2 (sodium azide and histidine) and free radical scavengers (cysteine) before the PDT treatment demonstrated the role of the ROS in the observed photo-cytotoxicity.

Acknowledgments: Thanks are due to the Universities of Aveiro and Coimbra, FCT and FEDER for funding the QOPNA and IBILI Units and the projects PTDC/CTM/101538/2008, PEst-C/SAU/UI3282/2011. Thanks to ACIMAGO (Ref. 12/12). P. Pereira and S. Silva thank to FCT for their BI (BI/UI55/5457/2011) and post-doctoral (SFRH/BPD/64812/2009) grants, respectively. References [1] Allison, R. R.; Sibata, C.H. Photodiagn. Photodyn. 2010, 2, 61. [2] Plaetzer, K.; Krammer, B.; Berlanda, J.; Berr, F.; Kiesslich, T. Lasers. Med. Sci. 2009, 2, 259. [3] Agostinis, P.; Berg, K.; Cengel, K. A.; Foster, T. H.; Girotti, A. W.; Gollnick, S. O.; Hahn, S. M.;

Hamblin, M. R.; Juzeniene, A.; Kessel, D.; Korbelik, M.; Moan, J.; Mroz, P.; Nowis, D.; Piette, J.; Wilson, B. C. and Golab, J. CA-Cancer J. Clin. 2011, 4, 250.

[4] Silva, S.; Pereira, P. M.; Silva, P.; Paz, F. A.; Faustino, M. A.; Cavaleiro, J. A. S.;Tome, J. P. C. Chem. Commun. 2012, 30, 3608.

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Novel water-soluble phosphonate phthalocyanines for the preparation of metal-organic frameworks

N. Venkatramaiah,a,b Filipe A. Almeida Paz,b João P. C. Toméa

aDepartment of Chemistry, QOPNA, University of Aveiro, 3810-193 Aveiro, Portugal; bDepartment of

Chemistry, CICECO, University of Aveiro, 3810-193 Aveiro, Portugal.

Water-soluble phthalocyanines (Pcs) have been shown to be effective photosensitizers (PSs) in photodynamic therapy (PDT) of cancer and photo inactivation of several micro- organisms and viruses in many infections or stored natural fluids.[1,2] Among the water-soluble Pcs, sulfonated, carboxylic and pyridinium derivatives have received great attention due to their accessibility.[3] The development of new photoactive molecules/materials, especially with different properties/functionalities, has engaged many synthetic chemists in recent years. The photophysical and chemical properties and their phototoxicity mechanisms of action are key factors of these molecules being dependent of the composition and structure of the PSs. In particular, the efficiency of generating cytotoxic active oxygen species remains a challenge.[4]

Synthesis of Pcs bearing phosphonate groups directly bound to the macrocycle core represents a novel area of research. These compounds should have high solubility in water being, thus, versatile building blocks for the construction of Metal-Organic Frameworks (MOFs). The P═O groups of phosphonates can play a vital role in different functions, as they are known to be strong hydrogen bond acceptors and exhibit interesting binding properties with saccharides. We have prepared a series of tetraphosphonated Pcs (1), both in the free-base form and having different central metal atoms. The synthesis, characterization and photophysical and photo chemical properties with particular emphasis on the role of these molecules in waste water treatment as photo catalysts is presented.

Acknowledgments: Thanks are due to the University of Aveiro, FCT and FEDER for funding the QOPNA and CICECO (PEst-C/CTM/LA0011/2011) research units and R&D project PTDC/QUI-QUI/098098/2008. N. Venkatramaiah is thankful to FCT for his post-doctoral grant (SFRH/BPD/79000/2011). References [1] Silva, S.; Patrícia, M. R. P.; Silva, P.; Paz, F.A.A.; Faustino, M.A.F.; Cavaleiro, J. A. S.; Tomé, J.P.C.

Chem. Commun. 2012, 48, 3608. [2] Pereira, J.B.; Carvalho, E.F.A.; Neves, M.G.P.M.S.; Cavaleiro, J. A. S.; Cunha, Â.; Almeida, A.; Tomé,

J.P.C. Photochem. Photobiol. 2012, 88, 537. [3] Maria C. D.; Crutchley, R.J. Coord. Chem. Rev. 2002, 233-234, 351. [4] Zhang, X. F.; Guo, W. J. Phys. Chem. A. 2012, 116, 7651.

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Comparative study of the interaction between α- and β-lapachone with bovine serum albumin (BSA)

Otávio Augusto Chaves,a,c Eduardo Benes,b Edgar Schaeffer,c Bauer de Oliveira

Bernardes,c Aurélio B.B. Ferreira,c José Carlos Netto-Ferreira,c Dari Cesarin-Sobrinhoc

aDepartment of Chemistry, University of Coimbra, 3004-535 Coimbra, Portugal; bDepartment of Chemistry, University Federal da Bahia, Ondina-Salvador, Brazil; cDepartment of Chemistry, University Federal Rural

do Rio de Janeiro, Seropédica, Brazil.

Serum Albumin is the most abundant protein in blood plasma and probably the most studied protein. Among its functions Serum Albumin are carriers, distributors and metabolizing agents of multiples ligands.[1] The lapachol α- and β-lapachone are naphthoquinones obtained from species of Tabebuia, exhibit anti-inflammatory, anti-bacterial, anticancer and trypanocidal properties. These substances and their derivatives have some application against tropical parasitic diseases, such as Chagas´disease.[2]

In order to understand the interaction between lapachones and BSA, we performed several experimental studies such as fluorescence, circular dichroism and UV-Vis spectroscopies. This study is important to understand the pharmacodynamics and pharmacokinetics in the distribution and elimination of the drugs in the body.

From the fluorescence data and applying the Stern-Volmer modified equation we obtained the values of the binding constants at temperatures of 288K, 293K and 298K. The observed Ka values are 106-107 L.mol-1, indicating a strong interaction between BSA and lapachones, mainly the α-lapachone. The high value of Kq (constant of fluorescence quenching) indicates that the process of fluorescence quenching of BSA is static (Kq = 2.20.1013L.mol-1).[3]

Using the values of ln Ka at these three temperature in a Van’t Hoff Stern-Volmer modified plot, we obtained the thermodynamic values ∆G°, ∆H° and ∆S°. The negative value of ∆G° to α-lapachone is consistent with the spontaneity of the binding, but the β-lapachone has a positive value of ∆G°. The positive value of ∆H° indicates that the binding process of both is endothermic, and the positive value of ∆S° shows that the type of interaction between the drugs with BSA is hydrophobic.[4]

The circular dichroism spectra indicate that the addition of the drugs influence the ellipticity of albumin, showing a decrease of two bands, 208nm and 222nm, mainly to α-lapachone, indicating that this one has more influence on the secondary structure of the protein. Acknowledgments: Thanks are due to the FAPERJ for the financial support and CAPES, for the international undergraduate scholarship (PLI/BEX4348/11-7). References [1] Galanos, C.; Rietshel, E.T.; Luderitz, O. European Journal of Biochemistry. 2005, 2, 230. [2] Netto-Ferreira, J.C.; Bernardes, A.B.B.; Ferreira, A.B.B. Chemistry and Physics. 2008, 10, 6645. [3] He, W.; Li, Y.; Tian, J.; Liu, H. Journal of Photochemistry and Photobiology. 2005, 174, 53. [4] Chen, G.Z.; Huang, X.Z.; Wang, Z.B. The Methods of Fluorescence Analysis. 1990, Science Press, 2sd

edition.

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Nitrogen heterocyclic compounds as acetylcholinesterase inhibitors: synthesis, bioactivity and docking studies

Maria do Carmo Barreto,a Diana C. G. A. Pinto,b Inês J. Sousa,c Miguel X. Fernandes,c

Artur M. S. Silvab

a CIRN/DCTD, Universidade dos Açores, 9501-801 Ponta Delgada, Portugal; b Department of Chemistry & QOPNA, University of Aveiro, 3810-193 Aveiro, Portugal; cCentro de Química da Madeira, Universidade da

Madeira, Campus Universitário da Penteada, 9000-390 Funchal, Portugal

Nitrogen heterocyclic compounds are one of the most important sources of new therapeutic agents. Furthermore, all the approved medications for Alzheimer’s disease are nitrogen-bearing molecules, most of which inhibit the enzyme Acetylcholinesterase (AChE), increasing the level of the neurotransmitter acetycholine, which is abnormally low in the brain of these patients.[1] However, the drugs currently used present severe side effects,[2] therefore the search for new compounds with less negative effects is of the utmost importance. In this context, we decided to synthesize nitrogen heterocyclic compounds, test their ability to inhibit AChE in vitro, and analyze the interaction between these molecules and the active center of the enzyme by computational docking studies.

In the present work, several nitrogen heterocycles were evaluated for their in vitro AChE inhibition by a modification of the Ellman method.[3] One of the compounds tested (1) was more active than galanthamine, an AChE inhibitor drug used in medicine, whilst the other three (2-4) presented lower activity, although comparable to galanthamine.

The interaction between the compounds and the active site of AChE was studied by computational docking, using the FlexScreen program, and the results were compared to biological data. The active site of AChE is a narrow gorge-like fold of the protein, and includes a peripheral site at the gorge mouth and a catalytic site, which in turn comprises an esteratic and an anionic site. For the studied compounds, the interactions were more favorable with the peripheral site. However, there was no relevant correlation between the energy values and the potency of inhibition. Possibly, the interaction between these compounds and the enzyme is exerted not only at the level of the active site but also in other parts of the protein molecule, and further studies are needed to fully characterize this interaction.

Acknowledgments: Thanks are due to the University of Aveiro, Fundação para a Ciência e a Tecnologia (FCT) and FEDER for funding the Organic Chemistry Research Unit (project PEst-C/QUI/UI0062/2011) and BIOPHARMAC-MAC/1/C104 and Project PEst-OE/QUI/UI0674/2011. Thanks are also due to CIRN (University of Azores) and DRCTC for funding the unit. References [1] Mehta, M.; Abdu Adem, A.; Sabbagh, M. Int. J. Alzheimers Dis. 2012, 728983.

doi:10.1155/2012/728983. [2] Patel, B.; Holland, N.W. Clinical Geriatrics 2011, 19, 27. [3] Arruda, M.; Viana, H.; Rainha, N.; Neng, N.R.; Rosa, J.S.; Nogueira, J.M.F.; Barreto, M.C. Molecules

2012, 3082.

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Protein-ligand docking study: diterpenes from Juniperus brevifolia as anticancer and antimicrobial agents

Inês J. Sousa,a Miguel X. Fernandes,a Ana M. L. Secab

aCentro de Química da Madeira, Campus da Penteada, University of Madeira, 9000-390 Funchal, Portugal;

bDCTD, University of Azores, 9501-801 Ponta Delgada,Portugal.

From leaves of Juniperus brevifolia, an endemic conifer from Azores, were isolated and structurally characterized, several dehydroabietane and sandaracopimarane derivatives.[1] Some of them (1-4), displayed antiproliferative activity against cancer cell lines (HeLa, A-549 and MCF-7) and bactericidal effect against Bacillus cereus at different concentrations tested.[2] However, it is not known how these compounds interact with most often proteins involved in the antimicrobial and cytotoxic mechanisms. Protein-ligand docking is mainly used to predict (energy and conformation wise) how small molecules bind to a protein of known 3D structure and to predict possible molecular targets for a set of compounds. In this work, the docking studies were performed, using the FlexScreen program, in order to pick molecular targets from a large set of common anticancer (63) and antimicrobial (39) targets to the selected compounds 1-4. The predicted interactions established between the compounds under study and the anticancer targets revealed that the compounds 1 and 3 interact preferentially with phosphatidylinositol-3,4,5-trisphosphate 5-phosphatase 2, whereas compounds 2 and 4 interact preferentially with human mitochondrial peptide deformylase and α-tubulin, respectively. Studying the interactions between the compounds 1 and 3 and the antimicrobial targets we predict that these compounds interact preferentially with RNA polymerase and peptide deformylase. These results provide additional understanding of the cytotoxic and antimicrobial effects of diterpenes studied. These preliminary computational docking predictions of therapeutic targets were established working with just 4 compounds, and to obtain more reliable predictions the number of compounds needs to be increased.

Acknowledgments: Thanks are due to the University of Azores, FCT, FEDER, BIOPHARMAC - MAC/1/C104 and Project PEst-OE/QUI/UI0674/2011. References [1] Seca, A. M. L.; Silva, A. M. S.; Bazzocchi, I. L.; Jimenez, I. A. Phytochemistry 2008, 69, 498. [2] Moujir, L. M.; Seca, A. M.L.; Araújo, L.; Silva, A. M.S.; Barreto, M. C. Fitoterapia 2011, 82, 225.

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Is Cytisus multiflorus an anti-inflammatory plant?

Susana C. Saraiva,a Olívia R. Pereira,a,b Joana Liberal,d,e Maria T. Batista,c,d Maria T. Cruz,c,e Susana M. Cardosoa

aCERNAS, Escola Superior Agrária de Coimbra, Instituto Politécnico de Coimbra, Coimbra, Portugal;

bDepartamento de Tecnologias de Diagnóstico e Terapêutica, Escola Superior de Saúde, Instituto Politécnico de Bragança, Bragança, Portugal; cFaculdade de Farmácia, Universidade de Coimbra,

Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal; dCentro de Estudos Farmacêuticos, Azinhaga de santa Comba, 3000-548 Coimbra, Portugal; eCentro de Neurociências e Biologia Celular, Departamento de

Zoologia da Universidade de Coimbra, 3004-517 Coimbra, Portugal

Cytisus multiflorus is a leguminous shrub native from Iberian Peninsula that is distributed in the south-west Mediterranean region. This plant is used in folk medicine and it is claimed to have various health benefits, including anti-inflammatory properties.[1] Yet, the anti-inflammatory usage of C. multiflorus is totally based on the available ethnopharmacological information while no scientific data on this capacity and on molecular targets has been reported for the plant. Hence, the present work aims to clarify the possible anti-inflammatory mechanisms of C. multiflorus.

A purified ethanolic extract was prepared and its high antioxidant capacity was confirmed though the DPPH radical scavenging[2] and reducing power[3] assays (EC50 values 13.4±1.0 and 11.4±2.1 µg/mL, respectively). Moreover, anti-inflammatory properties of the C. multiflorus extract were tested on a lipopolysaccharide-stimulated Raw 264.7 macrophages model. In order to accomplish that, nitric oxide (NO) production, scavenging activity and cytotoxicity of the extract were assessed. Furthermore, the effects on two proteins that are potential targets to prevent or treat chronic inflammation, namely cyclooxygenase-2 (COX-2) and inducible NO synthase (iNOS), were estimated by Western Blot analysis.

The obtained results showed that C. multiflorus extract was able to significantly inhibit the production of NO for non-toxic concentrations. The treatment of this cell line with 161 µg/ml and 325 µg/ml of the purified extract induced a decrease in the levels of NO of 24% and 32%, respectively. Furthermore, despite no changes on COX-2 levels were observed, iNOS expression was significantly diminished by the treatment with the highest concentration of the extract.

Overall, the present results suggest that C. multiflorus actually exerts an anti-inflammatory action which is, at least partially, mediated through the inhibition of iNOS expression.

Acknowledgments: The authors acknowledge the financial support provided by the FCT to CERNAS (project PEst-OE/AGR/UI0681/CNC/CEF/2011). Olívia R. Pereira was supported by a PhD grant (SFRH/PROTEC/49600/2009). References [1] Pereira, O. R.; Silva, A.M.S.; Domingues, M.R.M.; Cardoso, S.M.. Food Chem. 2012, 131, 652. [2] Ferreira, A.; Proença, C.; Serralheiro, M.L.M.; Araújo, M.E.M. J. Ethnopharmacol., 2006, 108, 31. [3] Oyaizu, M. Jpn. J. Nutr. 1986, 44, 307.

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Phenolic composition of Leonurus cardiaca L.

Olívia R. Pereiraa,b, Maria R. M. Domingues,c Susana M. Cardosoa

aCERNAS - Escola Superior Agrária, Instituto Politécnico de Coimbra, Coimbra, Portugal; bDepartamento de Tecnologias de Diagnóstico e Terapêutica, Escola Superior de Saúde, Instituto Politécnico de Bragança,

Bragança, Portugal; cDepartamento de Química & QOPNA, Universidade de Aveiro, Aveiro, Portugal.

Leonurus cardiaca L., also known as motherwort, is a plant native from central Europe that it is spread in different temperate countries around the world. The plant has been described to exert several beneficial properties and it is presently used in infusions, decoctions, syrups and tinctures, as well as an ingredient of pharmaceutical formulations.[1,2,3] Yet, the plant phenolics, which are frequently associated to the claimed health benefits of Leonurus cardiaca L., are not fully clarified. This topic is focused in the present work.

For that, an extract of the aerial parts of the plant was obtained with aqueous ethanolic solution (80%) and it was further purified onto SPE C18-E cartridges. The resulting purified phenolic extract was analyzed by HPLC-DAD and ESI-MSn techniques, for phenolic identification and quantification.

Accordingly, phenolic compounds in Leonurus cardiaca L. purified ethanolic extract accounted for 15 mg/g of dry plant (500 ± 49 mg/g of extract). Phenylethanoid glycosides were the most prevalent phenolics with lavandulifolioside and verbascoside representing 50% and 27% of its total quantified phenolic compounds, respectively. Additionally, the present study allowed the identification and quantification of leucoseptoside A and leonoside B for the first time in Leonurus cardiaca L., which together, accounted for 11% of the quantified phenolics. Besides phenylethanoid glycosides, the Leonurus cardiaca L. ethanolic extract also contained flavonoids (10%) and caffeic acid derivatives, albeit the latter were only present in vestigial amounts. With the exception of luteolin-7-O-rutinoside, all the remaining flavonoids were glycosidic quercetin derivatives, which include the isoquercitrin, rutin, rutin-O-glucoside and quercetin-3-O-sophoroside. Possible association of the major phenolics herein identified with specific health benefits of the plant are under investigation. Acknowledgments: The authors acknowledge the financial support provided by the FCT to CERNAS (project PEst-OE/AGR/UI0681/2011) and of the FCT as well as FSE (III Quadro Comunitário de Apoio) to QOPNA (project PEst-C/QUI/UI0062/2011), REDE/1504/REM/2005 (that concerns the Portuguese Mass Spectrometry Network). Olívia R Pereira was supported by a PhD grant (SFRH/PROTEC/49600/2009). References [1] Matkowski, A., Tasarz, P., Szypula, E. J. Med. Plants Res. 2008, 2, 321. [2] Jafari, S., Moradi, A., Salaritabar, A., Hadjiakhoondi, A., Khanavi, M. Res. J. Biol. Sci. 2010, 5, 484. [3] Xin-Hua, L., Hong, X., Yi-Zhun, Z. Acta Phys. Sin. 2007, 59, 578.

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New emissive porphyrins containing a coumarin unit selective for Hg(II) detection: Fluorescence studies

Carla I. M. Santos,a,b José Menezes,a Elisabete Oliveira,b,c M. Amparo F. Faustino,a Vitor F. Ferreira,d Fernando de C. da Silva,d José A. S. Cavaleiro,a M. Graça P. M. S. Neves,a

Carlos Lodeirob

aDepartment of Chemistry & QOPNA, University of Aveiro, 3810-193 Aveiro, Portugal; bBIOSCOPE Group, Faculty of Science, Physical-Chemistry Department, Ourense Campus, University of Vigo, 32004 Ourense, Spain; cVeterinary Science Department, (CECAV), University of Trás-os-Montes and Alto Douro, 5001-801 Vila Real, Portugal; dDepartamento de Química Orgânica, Universidade Federal Fluminense, 24020-141

Niterói, Rio de Janeiro, Brazil

The presence of mercury(II) in water is a serious environmental problem that has

been approached by many researchers.[1] During the last years many sensitive and accurate analytical techniques have been tested in the detection of this metallic ion, such as NMR spectroscopy, atomic absorption and mass spectrometry, but most of these detection techniques are sample-destructive methods.[2] Of the non-destructive-sample analytical methodologies, the most important are those based on optical devices such as chromogenic sensors or fluorescent chemosensors.[3]

As a part of our research project on the synthesis and characterization of new emissive chemosensors for metallic and anionic species, here we present two porphyrin derivatives containing a coumarin unit (L1-L2)

[4] that recognized, via absorption and emission spectroscopy the presence of Hg(II). The coordination behaviour toward the metal ions Zn(II), Pb(II), Cd(II), Fe(II), Ca(II), Na(I) and Ag(I) was also studied in solution, but a clear selective response towards Hg(II) was observed in all cases.

Fig. 1. Chemical structure of ligands L1 and L2, and bar chart of L1 in the presence of 2 equiv. of each metal ion.

Acknowledgments: Authors are grateful to Xunta de Galicia (Spain) project 09CSA043383PR and Scientific PROTEOMASS Association (Ourense-Spain) for financial support. Thanks are due to FCT-MEC and FEDER for funding the QOPNA unit (project PEst-C/QUI/UI0062/2011). C.S. and E.O thank also to FCT-MEC (Portugal) by their doctoral and Post-Doctoral grants, SFRH/BD/64155/2009 and SFRH/BPD/72557/2010, respectively. C.L. thanks Xunta de Galicia for the Isidro Parga Pondal Research Program. References [1] Liu, H.; Yu. P; Du, C.; He, B.; Qiu, X.; Chen, G. Talanta, 2010, 81, 433. [2] (a) Baumann, T. F.; Reynolds, J. G. Chem. Commun.1998, 16, 1637. (b) Nelson, A. J.; Reynolds, J. G.;

Baumann, T. F.; Fox, G. A. Appl. Surf. Sci. 2000, 167, 205. (c) Baumann, T. F.; Reynolds, J. G.; Fox, G. A. React. Funct. Polym. 2000, 44, 111.

[3] Mamelli, M.; Capelo, J.; Lodeiro, C. Inorg. Chem., 2010, 49, 8276. [4] Menezes, J. C. J. M. D. S.; Gomes, A. T. P. C.; Silva, A. M. S.; Faustino, M. A. F.; Neves, M. G. P. M.

S.; Tomé, A. C.; da Silva, F. C.; Ferreira, V. F.; Cavaleiro, J. A. S. Synlett, 2011, 13, 1841.

0

0 ,5

1

1 ,5

I/I0(a.u.)

Authors Index

AUTHORS INDEX

Auth

ors

Index

63

Alkorta, I. ......................................................................................................................................... 47 Almeida Paz, F. A. ........................................................................................................................... 54 Amslinger, S. .................................................................................................................................... 33 Anwar, A. ....................................................................................................................... 29, 30, 39, 40 Arbach, M. ................................................................................................................................. 30, 39 Asali, I. A. ........................................................................................................................................ 38 Bagrel, D. ................................................................................................................................... 41, 43 Bana, E. ............................................................................................................................................ 43 Barata, J. F. B. ............................................................................................................................ 51, 52 Barreto, M. C. .................................................................................................................................. 56 Batista, M. T. ................................................................................................................................... 58 Batteux, F. ........................................................................................................................................ 40 Benes, E. .......................................................................................................................................... 55 Bernardes, B. O. ............................................................................................................................... 55 Block, E. ........................................................................................................................................... 13 Brodziak-Jarosz, L. .......................................................................................................................... 33 Burkholz, T. ..................................................................................................................................... 25 Cardoso, S. M. ............................................................................................................................ 58, 59 Cavaleiro, J. A. S. ............................................................................ 15, 42, 45, 48, 49, 51, 52, 53, 60 Cerella, C. ........................................................................................................................................ 41 Cesarin-Sobrinho, D. ........................................................................................................................ 55 Chaves, O. A. ................................................................................................................................... 55 Chéreau, C. ....................................................................................................................................... 40 Claramunt, R. M. .............................................................................................................................. 47 Cruz, M. T. ....................................................................................................................................... 58 Czepukojc, B. ................................................................................................................................... 25 Dicato, M. ........................................................................................................................................ 41 Dick, T. ............................................................................................................................................ 33 Diederich, M. ............................................................................................................................. 24, 41 Domingues, M. R. M. ....................................................................................................................... 59 Eggleton, P. ...................................................................................................................................... 40 Elguero, J. ........................................................................................................................................ 47 Faustino, M. A. F. ...................................................................................................................... 51, 60 Fernandes, M. X. ........................................................................................................................ 56, 57 Fernandes, R. .................................................................................................................................... 53 Ferreira, A. B. B. .............................................................................................................................. 55 Ferreira, V. F. ................................................................................................................................... 60 Gaascht, F. ........................................................................................................................................ 41 Gerhäuser, C. .................................................................................................................................... 33 Glass, R. S. ....................................................................................................................................... 19 Groom, M. ...................................................................................................................... 13, 29, 30, 39 Hamilton, C. ......................................................................................................................... 26, 30, 39 Isupov, M. ........................................................................................................................................ 44 Jacob, C. ............................................................................................................. 25, 31, 32, 37, 38, 43 Jain, M. ............................................................................................................................................. 41 James, P. ........................................................................................................................................... 44 Jamier, V. ......................................................................................................................................... 40 Kavian, N. ........................................................................................................................................ 40 Kijjoa, A. .......................................................................................................................................... 17 Kirsch, G. ................................................................................................................. 31, 32, 34, 41, 43 Liberal, J. ......................................................................................................................................... 58 Littlechild, J. .................................................................................................................................... 44

Auth

ors

Index

64

Lo Faro, M. L. .................................................................................................................................. 36 Lodeiro, C. ................................................................................................................................. 51, 60 Lopes, G. R. ..................................................................................................................................... 50 Mai, A. ....................................................................................................................................... 18, 43 Marut, W. ......................................................................................................................................... 40 Meiser, P. ................................................................................................................................... 31, 43 Menezes, J. ....................................................................................................................................... 60 Montenarh, M............................................................................................................................. 37, 38 More, J. ............................................................................................................................................ 36 Nagy, P. ............................................................................................................................................ 14 Netto-Ferreira, J.-C. ......................................................................................................................... 55 Neves, M. G. P. M. S. .................................................................................................... 16, 51, 52, 60 Nicco, C. .......................................................................................................................................... 40 Nwachukwu, I. D. ............................................................................................................................ 35 Oliva, C. G. ...................................................................................................................................... 42 Oliveira, E. ....................................................................................................................................... 60 Panning, J. ........................................................................................................................................ 32 Pereira, O. R. .............................................................................................................................. 58, 59 Pereira, P. M. R. ............................................................................................................................... 53 Pinto, D. C. G. A. ............................................................................................... 15, 32, 45, 48, 50, 56 Pinto, J. ............................................................................................................................................. 47 Resende, D. I. S. P. ........................................................................................................................... 42 Reviriego, F. ..................................................................................................................................... 47 Ribeiro, C. A. F. ............................................................................................................................... 53 Rocha, D. H. A. ................................................................................................................................ 45 Saidu, N. E. B. ............................................................................................................................ 37, 38 Santos, C. I. M............................................................................................................................ 51, 60 Santos, C. M. M. .............................................................................................................................. 15 Santos, S. M. .................................................................................................................................... 52 Sanz, D. ............................................................................................................................................ 47 Saraiva, S. C. .................................................................................................................................... 58 Schaeffer, E. ..................................................................................................................................... 55 Schneider, T. .................................................................................................................................... 25 Schumacher, M. ............................................................................................................................... 41 Seca, A. M. L. .................................................................................................................................. 57 Seixas, R. S. G. R. ............................................................................................................................ 48 Servettaz, A. ..................................................................................................................................... 40 Sheridan, R. ...................................................................................................................................... 13 Silva, A. F. F. ................................................................................................................................... 52 Silva, A. M. G. ................................................................................................................................. 52 Silva, A. M. S. ................................................................ 15, 32, 42, 43, 45, 46, 47, 48, 49, 50, 52, 56 Silva, F. C. ........................................................................................................................................ 60 Silva, S. ............................................................................................................................................ 53 Silva, V. L. M. ............................................................................................................................ 47, 49 Slusarenko, A. J. ........................................................................................................................ 23, 35 Sousa, I. J. .................................................................................................................................. 56, 57 Talhi, O. ..................................................................................................................................... 32, 50 Teiten, M.-H. .................................................................................................................................... 41 Tomé, A. C. ...................................................................................................................................... 52 Tomé, J. P. C. ............................................................................................................................. 53, 54 Tomé, S. M. ...................................................................................................................................... 46 Torralba, M. C. ................................................................................................................................. 47 Torres, M. R. .................................................................................................................................... 47

Auth

ors

Index

65

Touma, R. ......................................................................................................................................... 38 Valente, S. ............................................................................................................................ 31, 34, 43 Venkatramaiah, N. ........................................................................................................................... 54 Viswanathan, U. ............................................................................................................................... 25 Wang, K. .......................................................................................................................................... 13 Weill, B. ........................................................................................................................................... 40 Whatmore, J. .................................................................................................................................... 36 Whiteman, M. .................................................................................................................................. 36 Winyard, P. G. ............................................................................................................................ 36, 40 Wolf, C. R. ....................................................................................................................................... 33 Wood, M. E. ..................................................................................................................................... 36 Xu, Z. ......................................................................................................................................... 31, 43 Zhang, S. .......................................................................................................................................... 13 Zwergel, C. ........................................................................................................................... 32, 34, 43

Participants List

PARTICIPANTS LIST

Part

icip

ants

Lis

t

69

Alan Slusarenko

alan.slusarenko@bio3.rwth-aachen.de

RWTH Aachen University,

Aachen, Germany

Ana Mafalda Vaz Martins

Pereira

mafaldapereira@ua.pt

QOPNA, University of Aveiro

Ana Maria Loureiro da Seca anaseca@uac.pt University of Azores

Ana Teresa Peixoto de Campos

Gomes

ana.peixoto@ua.pt

QOPNA, University of Aveiro

Anake Kijjoa ankijjoa@icbas.pt Universidade do Porto

Andreia Filipa Ferreira da Silva affs@ua.pt QOPNA, University of Aveiro

Andreia Sofia Filipe Farinha andreia.farinha@gmail.com QOPNA, University of Aveiro

Antonello Mai

antonello.mai@uniroma1.it

Sapienza University of Rome,

Rome, Italy

Artur Manuel Soares da Silva artur.silva@ua.pt QOPNA, University of Aveiro

Awais Anwar

awais.anwar@uea.ac.uk

ECOspray Limited, Suffolk,

United Kingdom

Carla Isabel Madeira dos

Santos

csantos420@gmail.com

QOPNA, University of Aveiro

Carla Patrícia Fernandes

Pereira

carlaf_pereira@hotmail.com

QOPNA & CICECO,

University of Aveiro

Carlos José Vieira Simões csimoes@qui.uc.pt University of Coimbra

Chris Hamilton

c.hamilton@uea.ac.uk

University of East Anglia,

Norwich, United Kingdom

Cláudia Marisa Barreiros

Neves

claudianeves@ua.pt

QOPNA, University of Aveiro

Claus Jacob

c.jacob@mx.uni-saarland.de

Saarland University,

Saarbrücken, Germany

Clemens Zwergel

clemens.zwergel@me.com

Université de Lorraine, Metz,

France

Clementina Maria Moreira dos

Santos

clems@ipb.pt

Instituto Politécnico de

Bragança

Cornelia Koeck

c.koeck@eurice.eu

Saarland University,

Saarbrücken, Germany

David Sadler-Bridge

ecospray@ecospray.com

ECOspray Ltd, Suffolk ,

Suffolk, United Kingdom

Part

icip

ants

Lis

t

70

Diana Cláudia Gouveia Pinto diana@ua.pt QOPNA, University of Aveiro

Diana Isabel Soares Pereira

Resende

dianaresende@ua.pt

QOPNA, University of Aveiro

Djenisa Helene Ascenção

Rocha

djenisa@ua.pt

QOPNA, University of Aveiro

Eduarda Marlene Peixoto da

Silva

eduarda.silva@ua.pt

QOPNA, University of Aveiro

Eric Block

eblock@albany.edu

University of Albany, Albany,

New York, USA

Flávio Alberto da Silva

Figueira

figueiraflavio@gmail.com

QOPNA, University of Aveiro

Francois Gaascht

francois.gaascht@lbmcc.lu

Recherches Scientifiques

Luxembourg

Frederic Batteux

batteux@gmail.com

Université Paris Descartes,

Hopital Cochin, Paris, France

Frederico Ribeiro Baptista ferdyribeiro@gmail.com QOPNA, University of Aveiro

Gabriella Costa

gabriellacostagc@gmail.com

Universidade Estadual de

Goiás, Goiás , Brazil

Gilbert Kirsch

kirsch@univ-metz.fr

Université de Lorraine, Metz,

France

Guido Rocha Lopes guido@ua.pt QOPNA, University of Aveiro

Gustavo Adolfo Lopes Ferreira

da Silva

gustavoasilva@ua.pt

QOPNA, University of Aveiro

Hélio Miguel Teixeira

Albuquerque

helio.albuquerque@ua.pt

QOPNA, University of Aveiro

Ifeanyi Daniel Nwachukwu

nwachukwu@bio3.rwth-aachen.de

RWTH, Aachen University,

Aachen, Germany

Inês Camoiana de Sousa

Cardoso

inescamoiana.s.cardoso@gmail.com

QOPNA, University of Aveiro

Isabel Cristina Maia da silva

Santos Vieira

ivieira@ua.pt

QOPNA, University of Aveiro

Jennifer Littlechild

J.A.Littlechild@exeter.ac.uk

University of Exeter, Exeter,

United Kingdom

Part

icip

ants

Lis

t

71

Joana Filipa Brites Barata

jbarata@dq.ua.pt

QOPNA & CICECO,

University of Aveiro

Joana Filipa Gonçalves Pinto jfpinto@ua.pt QOPNA, University of Aveiro

Joana Isabel Torrão da Costa jitc@ua.pt QOPNA, University of Aveiro

Joana Lia Cardoso de Sousa joanasousa@ua.pt QOPNA, University of Aveiro

Joana Patrícia Araújo Ferreira jpferreira@ua.pt QOPNA, University of Aveiro

Joana Teles Ferreira joanatferreira@ua.pt QOPNA, University of Aveiro

João Manuel Marques

Rodrigues

jrodrigues@ua.pt

QOPNA, University of Aveiro

João Paulo Costa Tomé jtome@dq.ua.pt QOPNA, University of Aveiro

José Carlos Joaquim Maia De

Souza Menezes

josemenezes@ua.pt

QOPNA, University of Aveiro

Leandro Miguel de Oliveira

Lourenço

leandrolourenco@ua.pt

QOPNA, University of Aveiro

Lidia Brodziak-Jarosz

l.brodziak@dkfz.de

German Cancer Research

Centre (DKFZ), Heidelberg,

Germany

Liliana Neto Costa liliana.neto@ua.pt QOPNA, University of Aveiro

Marc Diederich

marc.diederich@lbmcc.lu

Fondation de Recherche Cancer

et Sang, Luxembourg

Maria Clara Ferreira de

Almeida Cardia Gomes

clara.gomes@ua.pt

QOPNA, University of Aveiro

Maria da Graça de Pinho

Morgado Silva Neves

gneves@ua.pt

QOPNA, University of Aveiro

Maria do Amparo Ferreira

Faustino

faustino@ua.pt

QOPNA, University of Aveiro

Maria do Carmo Barreto barreto@uac.pt Universidade dos Açores

Maria Fernanda de Jesus Rego

Paiva Proença

fproenca@quimica.uminho.pt

University of Minho

Maria Letizia Lo Faro

maria.lofaro@pcmd.ac.uk

University of Exeter Medical

School, Exeter, United

Kingdom

Part

icip

ants

Lis

t

72

Mário Manuel Quialheiro

Simões

msimoes@dq.ua.pt

QOPNA, University of Aveiro

Mathias Montenarh

m.montenarh@mx.uni-saarland.de

Saarland University,

Saarbrücken, Germany

Miriam Arbach

m.arbach@uea.ac.uk

University of East Anglia,

Norwich, United Kingdom

Mónica Raquel Caseiro

Fernandes

monica.caseiro@ua.pt

QOPNA, University of Aveiro

Murree Groom

Groom@ecospray.com

ECOspray Ld, Suffolk ,United

Kingdom

Nathaniel E. B. Saidu

s9nasaid@stud.uni-saarland.de

University of the Saarland

(UdS), Homburg, Germany

Nuno Miguel Malavado Moura nmoura@ua.pt QOPNA, University of Aveiro

Olívia Rodrigues Pereira oliviapereira@ipb.pt Instituto Politécnico Bragança

Otavio Augusto Chaves

otavio_ufrrj@hotmail.com

University of Coimbra and

University Federal Rural do Rio

de Janeiro, Rio de Janeiro,

Brazil

Oualid Talhi oualid.talhi@ua.pt QOPNA, University of Aveiro

Patricia Luisa de Souza Bergo

patriciabergo@gmail.com

Universidade Federal de São

Carlos, Sao Paulo, Brazil

Patrícia Manuela Ribeiro

Pereira

patriciamanuela@ua.pt

QOPNA, University of Aveiro

Paul Winyard

paul.winyard@pms.ac.uk

University of Exeter, Exeter,

United Kingdom

Pedro António Martins Mira

Varandas

pedrovarandas@ua.pt

QOPNA, University of Aveiro

Peter Meiser

p.meiser@ursapharm.de

Ursapharm Arzneimittel

GmbH, Saarbrücken, Germany

Péter Nagy

peter.nagy@oncol.hu

The National Institute of

Oncology, Budapest, Hungary

Raquel González Soengas rsoengas@ua.pt QOPNA, University of Aveiro

Part

icip

ants

Lis

t

73

Raquel Sofia Grevy Ribeiro

Seixas

raquel.grevy@ua.pt

QOPNA, University of Aveiro

Richard Glass

rglass@u.arizona.edu

University of Arizona, Tucson,

Arizona, USA

Samuel Guieu

sguieu@ua.pt

QOPNA & CICECO,

University of Aveiro

Sara Mirassol Tomé smt@ua.pt QOPNA, University of Aveiro

Sergio Valente

sergio.valente1977@libero.it

Sapienza University of Rome,

Rome, Italy

Sónia Maria Gomes Pires sonia.pires@ua.pt QOPNA, University of Aveiro

Sónia Pereira Lopes sonialopes@ua.pt QOPNA, University of Aveiro

Stéphanie Branco Leal stephaniebranco@ua.pt QOPNA, University of Aveiro

Susana Costa Saraiva

susaraiva90@gmail.com

Escola Superior Agrária de

Coimbra

Susana Santos Braga sbraga@ua.pt QOPNA, University of Aveiro

Venkatramaiah Nutalapati

nvenkat83@gmail.com

QOPNA & CICECO,

University of Aveiro

Vera Lúcia Marques da Silva verasilva@ua.pt QOPNA, University of Aveiro

Vera Mónica Sousa Isca veraisca@ua.pt QOPNA, University of Aveiro

Vincent Jamier

vincent.jamier@icn.cat

Catalan Institute of

Nanotechnology, Bellaterra,

Spain

Vinicius de Oliveira Silva

viniciosoliver@hotmail.com

Universidade Estadual de

Goiás, Goiás , Brazil

Wioleta Marut

vmarut@gmail.com

Paris Descartes, Hopital

Cochin, Paris, France

Zhanjie Xu

z.xu@mx.uni-saarland.de

Ursapharm Arzneimittel

GmbH, Saarbrücken, Germany