2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
Area della Ricerca di Sassari, 8-9 Ottobre 2015
2nd Convention of the CNR Institute of Biomolecular Chemistry
Sassari Reasearch Area, October 8-9, 2015
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
Il comitato scientifico:
Nicola D’Antona
Giovanna Delogu
Luciano De Petrocellis
Corrada Geraci
Angela Patti
Paolo Ruzza
Daniele Sanna
Maurizio Solinas
Antonio Trincone
Rosa Maria Vitale
Il comitato organizzatore
Emanuela Azara
Antonella Dettori
Marina Pisano
Gloria Rassu
Maria Serra
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2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
“Biomolecular Chemistry” and the Italian Research Institute that has made a mission of it
When the CNR Institute of Biomolecular Chemistry (ICB) was founded in 2002, most of its
personnel hardly knew how to explain to the layman what this “new” discipline, Biomolecular
Chemistry, really was. Although obviously everybody knew what Chemistry is, and how it can be
elegantly applied to the understanding of the molecular mechanisms regulating biological systems
and processes (ICB was in fact founded starting from the National Coordination Institute for the
Chemistry of Biological Systems), the adjective “Biomolecular” seemed to define only a general,
and perhaps too vague, idea of what would have been the subjects of the studies of the new
institute. Yet, since from the first (and, until last year, only) general meeting of ICB, in 2002, it was
clear that researchers and technicians of the institute would have focused their research efforts on
the understanding of the multifaceted roles of biomolecules in all aspects of our life, and on their
technological application to respond to the ever increasing needs not only of the bio-medical field,
but also of the agricultural, environmental and renewable energy sectors. Those were also the
years of the rise of two new disciplines, Chemical Biology and Bioorganic Chemistry, which
seemed to be nicely tailored to what ICB researchers planned to work on.
In the meantime, the term “Biomolecular” has been increasingly used over the last 15 years.
Taking into account only the PubMed engine website, this word went from only 327 citations in
1996 to well above 5570 in 2014, thus indicating that its choice in 2002 had been both timely and
successful. “Biomolecular Sciences” is now a widespread discipline in Italian Universities, to the
point that, for example, doctorate courses on this specific subject have appeared in many faculties.
As a consequence, when I applied to become the Director of ICB, and since the beginning of my
mandate in June 2014, I thought that the mission of the Institute should continue to be the
“chemical study of biological systems and processes to understand their function at the molecular
level and explore their new technological applications” and “to operate in the fields of bioorganic
chemistry and chemical biology, with expertise in the disciplines of spectroscopy, structural and
synthetic chemistry, biochemistry, molecular modeling, microbiology, pharmacology, nutraceutics
and bioenergetics”. In other words, I strongly believed that ICB research activity should continue
to be centered on organic molecules and their biological properties.
Last October, researchers from the four ICB branches met again, in the Naples main branch of the
institute and for the first time in 12 years. During the course of two very intense days, they provided
an update of their current efforts towards fulfilling this mission, discussed their latest results, and
threw the bases for new intra-mural collaborations reflecting and exploiting the inter- and multidisciplinary nature of their expertise. This “experiment”, made necessary by the low degree (as
recognized by most of the personnel) of previous scientific contacts among researchers from the
different ICB branches, was very successful. It resulted in the forming of 5 “Inter-Section”
disciplinary groups and in the beginning of at least two new collaborations. The meeting also gave
momentum to the action of the 4 “Institute Services of Common Interest” (SICI), which are now
working very well to provide all sorts of information to ICB personnel on the subjects of their
competence, and to establish new contacts with the “outside world”.
From the scientific standpoint, the 2014 ICB meeting did not focus on any of the several
institutional topics of research in particular, as it aimed at providing ICB researchers with a general
view of the state of the art of all ICB experimental activities, “old” and “new”. For this year’s
meeting in Sassari, instead, the Scientific Committee has made a tremendous effort to select, out
of the many present in ICB, three main topics to focus the program on (Drug Discovery, Omics and
Green Chemistry), and have invited for each of these topics renowned scientists from outside the
institute and the CNR. This will increase the exchange of information between ICB and other
research institutions, and enhance further the visibility of our institute. At the same time, the
Committee have allowed for a wide participation of the institute’s research personnel to show the
results of their activities on all topics of interest of ICB in the form of posters, whose presentation
will be given equal importance as the oral presentations and over 3 hours for discussion.
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2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
The result is what I believe to be an excellent and exciting program, for which I am very grateful to
the Scientific Committee, as it is emblematic of the great variety of the research interests and
activities of ICB, and their potential utility to help solving some of the unmet demands of our
society.
I hope we will enjoy this year’s annual meeting at least as much as we did last year, and I am
confident that new collaborations, ideas and strategies will come out of these two days together.
These, in turn, will help us coping with the public funding and, particularly, human resources
difficulties that affect our institute as well as research in Italy in general, and, at the same time, will
contribute at making ICB our home and a prestigious and renowned research institute in both our
country and abroad.
Vincenzo Di Marzo
Direttore Istituto di Chimica Biomolecolare
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2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
Programma del 2° Convegno dell’Istituto CNR di Chimica Biomolecolare
8-9 Ottobre 2015, AULA MAGNA – Area di Ricerca CNR di Sassari
Giovedì 8 Ottobre
12:30–13:00
Messaggi di benvenuto del Direttore e del Responsabile UOS di Sassari – Ricordo dei colleghi Aniello Lopez
ed Ugo Anselmi
13:00–14:30 Lunch di benvenuto
PL1. 14:30–15:00
Moderatore: Giovanna Delogu
Alba Canu – Centro di Restauro della Soprintendenza Archeologia della Sardegna, Ministero per i Beni e le
Attività Culturali e il Turismo (MIBACT) – Sassari
Restoration, Archaeometry and diagnostic: an essential collaboration
OC1. 15:00–15:20
Pietro Spanu
Potent non-covalent inhibitors of Caspase-1 therapeutically targeting neuroinflammation in
neurodegenerative diseases
OC2. 15:20–15:40
Davide Fabbri
Sustainable coupling reactions of natural bioactive hydroxylated phenols
15:40–16:00 Coffee break
16:00–17:30 Poster session
17:30–19:00
Riunione Istituto di Chimica Biomolecolare
Moderatore: Vincenzo Di Marzo
CENA SOCIALE
Venerdì 9 Ottobre
PL2. 9:00–9:30
Moderatore: Giovanni Nicolosi
Anna Maria Orlandoni – Matrìca S.p.A. – Porto Torres (SS)
Third generation biorefineries: Matrìca case - Opportunity and new developments
OC3. 9:30–9:50
Fabiana Piscitelli
Endocannabinoidomics: “omics” approaches applied to endocannabinoids and endocannabinoid-like
mediators
OC4. 9:50–10:10
Paola Bureddu
Synthesis and characterization of aminoproline-based RGD semipeptides targeting V3 integrins and their
utility in medicine
OC5. 10:10–10:30
Maurizio Solinas
Screening of N,N-bidentate pyridine-based ligands in copper and palladium catalysed reactions
10:30–10:50 Coffee break
OC6. 10:50–11:10
Moderatore: Antonio Trincone
Barbara Biondi
Peptaibols in membrane-llike environments: use of nitroxyl-labeled peptides to get insight into mechanism of
action drug discovery&recognition
iv
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
OC7. 11:10–11:30 Rosaria Villano
Efficient and environmentally friendly methodologies for the synthesis of multifunctionalized scaffolds
OC8. 11:30–11:50 Giuseppe Granata
Assembly of a succinyl-calix[4]arene derivative in multilamellar vesicles
11:50–12:50 Poster session
12:50–14:00 Pranzo
PL3. 14:00–14:30
Moderatore: Andrea Calderan
Sebastiano Banni – Università degli Studi di Cagliari UNICA – Dipartimento di Biologia Sperimentale,
Sezione di Fisiologia Generale
Nutritional role of lipids and their potential use in functional foods and nutraceuticals
OC9. 14:30–14:50
Adele Cutignano
A 'sea' of bioactive natural products
OC10. 14:50–15:10
Nicola Culeddu
NMR metabolomic applications on olive oils: pedoclimatic and sensory differentiation. From single pulse
NMR to adiabatic sequence
PL4. 15:10–15:40
Moderatore: Mauro Marchetti
Pier Andrea Serra – Università degli Studi di Sassari UNISS – Dipartimento di Medicina Clinica e
Sperimentale, Sezione di Farmacologia
Electrochemical micro(bio)sensors as new analytical tools for Biomedical and Agri-Food applications
15:40–16:00 Coffee break
16:00–16:40 Poster session
OC11. 16:40–17:00
Carmelo Drago
One-pot microwave assisted catalytic transformation of waste frying oil into glycerol-free biodiesel
OC12. 17:00–17:20
Margherita Gavagnin
Carrying on the route of marine guanidines: isolation, synthesis and bioactivity of a guanidine terpene from
the mollusk Actinocyclus papillatus
17:20 Conclusioni
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2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
Relatori invitati – Invited speakers
“Restoration, Archaeometry and diagnostic: an essential collaboration”
Alba Canu – Centro di Restauro della Soprintendenza Archeologia della Sardegna,
Ministero per i Beni e le Attività Culturali e il Turismo (MIBACT) – Sassari
“Third generation biorefineries: Matrìca case - Opportunity and new developments”
Anna Maria Orlandoni - Matrìca S.p.A. – Porto Torres (SS)
“Nutritional role of lipids and their potential use in functional foods and nutraceuticals”
Sebastiano Banni – Università degli Studi di Cagliari UNICA
Dipartimento di Biologia Sperimentale, Sezione Fisiologia Generale
“Electrochemical micro(bio)sensors as new analytical tools for Biomedical and Agri-Food
applications”
Pier Andrea Serra – Università degli Studi di Sassari UNISS
Dipartimento di Medicina Clinica e Sperimentale, Sezione di Farmacologia
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2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
Oral Presentations
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
POTENT NON-COVALENT INHIBITORS OF CASPASE-1
THERAPEUTICALLY TARGETING NEUROINFLAMMATION IN
NEURODEGENERATIVE DISEASES
Ulgheri F.1, Deligia F.1, Morra A.1, Fuggetta M.P.2, Chandgude A. L.3,
Doemling A.3, Spanu P.1
1
Institute of Biomolecular Chemistry-CNR, Trav. La Crucca 3, 07100,Sassari,
Italy
2
Institute of Translational Pharmacology-CNR, Via Fosso del Cavaliere,00133,
Rome, Italy
3
Department of Drug Design, University of Groningen, Groningen, 9713 AV,
Netherlands
email: [email protected]
Recent studies have shown the pivotal role of innate immune activation in
the pathogenesis of major neurodegenerative diseases. It was discovered that
microglia and other cell types components of the brain innate immune system,
may be activated also by misfolded proteins or aberrant endogenous molecular
patterns that are accumulated in the brain of patients affected by several
neurodegenerative disorders. These stimuli are responsible for chronic
neuroinflammation by triggering the inflammasome's formation and subsequent
caspase-1 activation. Proinflammatory cytokines are processed by active caspase1 into their mature form that leads to cytokines production and, ultimately, chronic
inflammation. Furthermore, the extensive generation of proinflammatory cytokines
(IL-1β, IL-18) mediated from activated caspase-1, leads to a new process of
programmed cell death called pyroptosis that plays an important role in several
neurological diseases. Moreover, recent findings indicate caspase-1 as modulator
for the activation of caspase-6-mediated axonal degeneration in AD.
These findings have changed the view of the role of chronic
neuroinflammation in neurodegenerative diseases, showing that this could be the
primary cause of the neurodegeneration and not a simple effect of the pathologies.
Additionally, there is evidence of the key role of caspase-1 in neurodegeneration,
and its inhibition might effectively interfere with the progression of neurological
disorders. So, caspase-1 inhibitors are very promising candidates for the treatment
of neurodegenerative diseases. For this purpose, we have designed and
synthesized new potent and selective non-peptidic, non-covalent and cell
permeable caspase-1 inhibitors, which have shown a very high activity in
suppressing the formation of IL-1 in LPS induced inflammation.
Acknowlegment: Research funded by Regione Autonoma della Sardegna- Legge Regionale 7
References: a) Heneka, M. T.; Kummer, M. P.; Latz, E. Nat. Rev. Immunol. 2014, 14, 463–477. b)
Agyemang, A. F.; Harrison, S. R.; Siegel, R. M.; McDermott, M. F. Semin. Immunopathol. 2015, 37,
335-347. c) Kaushal, V.; Dye, R.; Pakavathkumar, P.; Foveau, B.; Flores, J.; Hyman, B.; Ghetti, B.;
Koller, B. H.; LeBlanc, a C. Cell Death Differ. 2015, 1–11.
Oral 1
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
SUSTAINABLE COUPLING REACTIONS OF NATURAL BIOACTIVE
HYDROXYLATED PHENOLS
Fabbri D., Dettori M. A., Delogu G.
CNR - Institute of Biomolecular Chemistry, Traversa La Crucca 3, I-07100 Sassari, Italy
email: [email protected]
Hydroxylated phenols like vanillin (4-hydroxy-3-methoxybenzaldehyde) 1, apocynin (1-(4hydroxy-3-methoxyphenyl)-ethanone
2,
iso-apocynin
1-(2-hydroxy-3methoxyphenyl)ethanone), 3 creosol (2-methoxy-4-methylphenol) 4, zingerone (4-(4hydroxy-3-methoxyphenyl)-2-butanone) 5 and eugenol (2-methoxy-4-(propen-2-yl)-phenol)
6, are naturally occurring compounds, present, as main components, in various plants like,
Vanilla planifolia, Picrorhiza kurroa, Guaiacum officinale, Zingiber officinalis and Eugenia
carofillata. These compounds show a broad spectrum of biological properties such as
antiinflammatory, antiseptic, antioxidant and fungicide.a,b The corresponding C2-symmetric
dimers, named hydroxylated biphenyls, generally show a higher bioactivity in comparison
to the monomers.c Our interest in the field of hydroxylated biphenyls pushed us towards
the optimization of oxidative coupling reactions of monomers 1-6 with particular attention
to aspects of environmental sustainability of the synthesis. The reactions reported in the
literature for C2 symmetric dimerization of phenols 1-6, contemplate reactions mediated by
chemical inorganic oxidants,d,e typically iron sulphate and potassium persulfate, or
enzymatic reactionsf (e.g. horseradish peroxidase). Surprisingly, to the best of our
knowledge, there are not reported oxidative coupling reactions carried out with the aid of
microwaves on natural phenols 1-6.
Dimers 7-12 are important precursors of most of bioactive hydroxylated biphenyls recently
prepared by our group.g-l Our work is therefore aimed at optimizing, with the aid of
microwaves, the synthetic process in terms of reduction of the amount of solvent,
exclusive use of water as reaction medium and simplification of work-up operations.
Acknowlegment: Research funded by: L.R. n.7 : "Promozione della ricerca scientifica e dell'innovazione
tecnologica in Sardegna”: crp-17136 “Salute & Trigu” 2012-1014, crp-25114 “Cosmesagro” 2012-1014 and
PRIN D.M. 1152/ric del 27/12/2011, “Cell wall determinants to improve durum wheat resistance to Fusarium
diseases”.
References: a) Kim, J-H. et al. Plos one, 2014, 9, 1-10. b) Lin, C-H. et al. Mol. Plant-Micr. Interact. 2009, 22,
942-952. c) Klees, R. et al. J. Biomed. Biotechnol. 2006, 2:87246. d) Forsythe, W. G. et al. Green Chem.
2013, 15, 3031-3038. e) Delomenede, M. et al. J. Med. Chem. 2008, 51, 3171-3181. f) Antoniotti, S. et al.
Org. Lett. 2004, 6, 1975-1978. g) Dettori, M.A. et al. Lett. Drug Des. Discov. 2015, 12, 131-139; h) Marchiani,
A. et al. Amino Acids, 2013, 45 (2), 327-338; i) Rozzo, C. et al. Mol. Cancer, 2013, 12:37. l) Pani, G. et al. J.
Agr. Food. Chem. 2014, 62 (22), 4969-4977.
Oral 2
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
ENDOCANNABINOIDOMICS: “OMICS” APPROACHES APPLIED
TO ENDOCANNABINOIDS AND ENDOCANNABINOID-LIKE
MEDIATORS
Piscitelli F. 1, Bisogno T.1, Giordano C.2, Luongo L.2, Maione S.2, Di
Marzo V.1
1
Institute of Biomolecular Chemistry, Consiglio Nazionale delle Ricerche, Via
Campi Flegrei 34, 80078, Pozzuoli (NA), Italy; 2 Department of Experimental
Medicine/Division of Pharmacology, Second University of Naples, via
Costantinopoli, 16 80138, Naples, Italy.
email: [email protected]
Endocannabinoids (ECs) are defined as the endogenous ligands of cannabinoid
receptors (CB1 and CB2) and a growing body of evidence has emerged on the
role of the endocannabinoid system (ECS) in the regulation of several
physiological conditions and numerous diseases. The development of analytical
techniques for endocannabinoid measurement in several biological matrices has
been helpful for the understanding of the physiological and pathological role of
these mediators. Moreover, since ECs and EC-like molecules are all derivatives of
fatty acids, several lipidomics approaches have been applied to date. The
application of targeted lipidomics methods to the analysis of ECs and EC-related
mediators is part of those methodologies used to investigate the
“endocannabinoidome” and defined as “endocannabinoidomics” and increasing
evidence has shown the importance of this strategy to identify new EC-like
molecules and define their biological role and metabolic pathways (a). In particular,
the application of “endocannabinoidomics” tecnique in a model of mouse traumatic
brain injury (TBI) led to the identification of a poorly investigated EC-related
molecule, the N-oleoylglycine (OlGly) in the injured brain. Therefore, to test the
anti-hyperalgesic and neuroprotective effects of OlGly mice underwent TBI using
the weight drop model and were divided into seven experimental groups: naïve,
sham, sham+OlGly 50 and 100 mg/kg, TBI, TBI+OlGly 50 and 100 mg/kg. Animals
were treated with OlGly (i.p.) for 14 days once a day, starting one day after injury
and underwent, several behavioural tests to assess pain and depression. At the
end of the treatment, the animals were decapitated and brains were dissected for
endocannabinoid analysis. Interestingly, treatment with OlGly normalized motor
impairment and reckless behavior; reduced thermal hyperalgesia and mechanical
allodynia and, moreover, normalized aggressiveness and depression induced by
TBI. On the other hand, the levels of endocannabinoids did not undergo any
change 65 days after TBI. In conclusions, the application of a very targeted
method for the discovery of new EC-like mediators is becoming necessary for the
understanding of their biological role and the development of new diagnostic
biomarkers and this study, described here, highlighted the neuroprotective role of
a new bioactive lipid in a model of brain injury.
References:
a) Piscitelli F. In: Di Marzo & Wang (Eds.), Elsevier Academic Press, London, 2015, 137-149
Oral 3
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
SYNTHESIS AND CHARACTERIZATION OF AMINOPROLINEBASED RGD SEMIPEPTIDES TARGETING V3 INTEGRINS AND
THEIR UTILITY IN MEDICINE
Burreddu P.1, Battistini L.2, Sartori A.2, Rassu G.1, Casiraghi G.2,
Zanardi F.2
1
Istituto di Chimica Biomolecolare del CNR, Traversa La Crucca 3, 07100 Li Punti,
Sassari, Italy. 2Dipartimento di Farmacia, Università degli Studi di Parma, V.le G.P.
Usberti 27A, 43124 Parma, Italy
email: [email protected]
In recent past years, integrin receptors have been the focus of intense research
directed to elucidate their structure, function and regulation.1 In the cancer-related
field, the expression of particular integrins is correlated with disease progression
and decreased patient survival in various tumor types, rendering these integrin
families appealing targets for cancer therapy. The V3 integrins, among others,
have been identified as useful biomarkers of tumor angiogenesis and tumor
progression, invasion and metastasis, being overexpressed on proliferating
endothelial cells as well as various tumor-related cells. A number of specific, highly
potent V3-targeting small molecule ligands have been developed so far, which
contain or mimic the essential RGD binding motif.2
Notable results from our laboratory in the design, synthesis and characterization of
aminoproline-based integrin binders (AmpRGD) will be presented, which displayed
nanomolar binding affinity toward the isolated V3 integrin receptor.3 The
preparation and evaluation of both covalent and nanostructured assemblies will
also be discussed, wherein appropriate cytotoxic or imaging cargos are consigned
to the AmpRGD semipeptide vectors, to be used as novel anti-angiogenic
therapeutic/diagnostic tools.
(1) Cox, D.; Brennan, M.; Moran, N. Nature Rev. Drug Discovery 2010, 9, 804-820.
(2) Auzzas, L.; Zanardi, F.; Battistini, L.; Burreddu, P.; Carta, P.; Rassu, G.; Curti, C.; Casiraghi, G.
Curr. Med.Chem. 2010 , 17, 1255-1299.
(3) (a) Battistini, L.; Burreddu, P.; Sartori, A.; Arosio, D.; Manzoni, L.;Luigi Paduano, L.; D’Errico, G.;
Sala, R.; Reia, L.; Bonomini, S.; Rassu, G.; and Zanardi, F. Mol Pharmaceutics, 2014, 11, 22802293. (b) Pilkington-Miksa, M.; Arosio, D.; Battistini, L.; Belvisi, L.; De Matteo, M.; Vasile, F.;
Burreddu, P.; Carta, P.; Rassu, G.; Perego, P.;Carenini, N.; Zunino, F.; De Cesare, M.; Castiglioni,
V.; Scanziani, E.; Scolastico, C.; Casiraghi, G.; Zanardi, F.; Manzoni, L. Bioconjugate Chem., 2012,
23, 1610-1622.
Oral 4
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
SCREENING OF N,N-BIDENTATE
PYRIDINE-BASED LIGANDS IN COPPER AND PALLADIUM
CATALYSED REACTIONS
Solinas M.
Istituto di Chimica Biomolecolare, UOS di Sassari
Traversa La Crucca, 3 – Regione Baldinca, Li Punti, 07100 Sassari
email: [email protected]
In the past few decades, a plethora of efficient chiral ligands has been developed
in several asymmetric reactions. Some of these ligands are mixed bidentate donor
ligands with P–P or P–N chelating mode, although several interesting examples of
N,N-chelating ligands such as diamines, pyridine-based ligands (2,2’-bipyridines,
1,10-phenanthrolines, pyridyl amines, pyridyl oxazolines, etc.) and oxazolinecontaining ligands have also proven their efficiency in several processes.
Moreover, chiral N,N-ligands based on imino and pyridine moieties have become a
large and important topic in asymmetric synthesis.
In our ongoing research efforts aimed at the application of chiral ligands based on
the pyridine framework to asymmetric catalysis, we present here the potential of a
variety of chiral N,N-bidentate ligands in the asymmetric Cu(I)-catalysed allylic
oxidation of cyclic alkene, in the Cu(II)-catalyzed Henry reaction and in the Pdcatalyzed allylic alkylation of 1,3-diphenylprop-2-enyl acetate with dimethyl
malonate.
Acknowlegment: Research funded by Fondazione Banco di Sardegna
References: a) Maurizio Solinas, Barbara Sechi, Salvatore Baldino, Giorgio Chelucci J. Mol. Catal.
A Chem. 378 (2013) 206–212. b) Maurizio Solinas, Barbara Sechi, Giorgio Chelucci, Salvatore
Baldino, José R. Pedro, Gonzalo Blay J. Mol. Catal. A Chem. 385 (2014) 73–77. c) Maurizio
Solinas, Barbara Sechi, Giorgio Chelucci Appl. Organometal. Chem. 2014, 28, 831–834.
Oral 5
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
PEPTAIBOLS IN MEMBRANE-LIKE ENVIRONMENTS: USE OF
NITROXYL-LABELED PEPTIDES TO GET INSIGHT INTO
MECHANISM OF ACTION
Biondi B.1, Bortolus M.2, Dalzini A.2, De Zotti M.2, Formaggio F.2,
Peggion C.2
1
Institute of Biomolecular Chemistry, Padova Unit, 35131, Padova
Department of Chemical Sciences, University of Padova, 35131, Padova
email: [email protected]
2
Peptaibols are a class of natural occurring antimicrobial peptides (AMPs),
characterized by a high content of the non-coded Cα-tetrasubstituted amino acid
Aib (α-aminoisobutyric acid), a C-terminal 1,2-amino alcohol, and acylated at the
N-terminus. Peptaibols are particularly attractive for their resistance to enzymatic
degradation and for a remarkable bioactivity, displayed even by short sequences,
due to the presence of Aib that favors very stable secondary structures. Like all
AMPs, peptaibols exert their antimicrobial action through a permeation of the
bacterial membranesa.
In this contribution, we report on the syntheses of various analogs of different
peptaibols in which the Aib residue is replaced with the achiral tetrasubstituted αamino
acid
4-amino-1-oxyl-2,2,6,6,-tetramethylpiperidine-4-carboxylic
acid
(TOAC). TOAC amino acid, which presents a stable nitroxyl radical fixed on a rigid
heterocyclic structure, can be used as paramagnetic probe to study, by electron
spin resonance (ESR), the mode in which peptides interact with a membrane b.
In the last decade, a great attention has been devoted to the use of AMPs as
anticancer agents. Despite differences in the sequences there are some common
characteristics: they are often positively charged and adopt an amphiphilic helical
structure in membrane environment. Most AMPs attack cancer cells by disrupting
the cell membrane without binding to a specific receptor, thus by-passing the
ability of cancer cells to adapt and developing treatment resistance. In this context,
we synthesized various analogs of the natural peptaibol trichogin GA IV c, some of
which bearing a nitroxyl probe at different positions, in order to evaluate the
selectivity of trichogin analogs toward tumor cells and correlate the variance in
bioactivity to different mechanism of peptide-membrane interaction.
Acknowlegment: CARIPARO Foundation (Progetto di Eccellenza 2011-12).
References:
a) Toniolo, C. and Brückner, H., Peptaibiotics, Wiley/VCH, Weinheim/ Zürich, 2009.
b) Marsh, D., Jost, M., Peggion, C., Toniolo, C. Biophys. J., 2007, 92, 473-481.
c) De Zotti, M.; Biondi, B.; Peggion, C.; Formaggio, F.; Park, Y.; Hahm, K.-S.; Toniolo C. Org. Biomol.
Chem., 2012, 10, 1285-1299.
Oral 6
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
EFFICIENT AND ENVIRONMENTALLY FRIENDLY
METHODOLOGIES FOR THE SYNTHESIS OF
MULTIFUNCTIONALIZED SCAFFOLDS
Villano R.
ICB-CNR, trav. La Crucca 3, 07100 Sassari
email: [email protected]
The vinylogous addition of Chan’s diene 11 to aldehydes is one of the most elegant
and powerful methods for the synthesis of δ-hydroxy-β-ketoesters 2 (Scheme 1)
which are useful key-intermediates in the synthesis of many natural products;2 so
the development of innovative methodologies for this reaction represents an
interesting research area.
Me3SiO
OH O
OSiMe3
OMe
+
RCHO
R
O
OMe
2
1
Scheme 1
The reaction has been intensely investigated over the last decade and particular
attention has recently focused on the use of low impact solvents and catalytic
amounts of promoters, according to the growing demand for highly sustainable
chemical procedures.
In the light of the above, an overview of several convenient and efficient
methodologies3 for the addition of Chan’s diene to aldehydes will be presented,
with particular attention to experimental conditions and catalytic systems (including
organometallic complexes and organocatalysts).
Finally, some applications of these methodologies in the total synthesis of complex
molecules will be discussed.
Acknowledgment: Research funded by Università degli Studi di Salerno
References:
1) a) Yamamoto, K.; Suzuki, S.; Tsuji, J. Chemistry Letters 1978, 649. b) Chan, T.-H; Brownbridge,
P. J. C. S. Chem. Comm. 1979, 578. c) Brownbridge, P.; Chan, T.-H; Brook, M. A.; Kang, G. J. Can.
J. Chem. 1983, 61, 688.
2) a) Evans, D.A.; Hu, H.; Burch, J.D.; Jaeschke, G. J. Am. Chem. Soc. 2002, 124, 5654. b)
Peterson, I.; Davies, R.D.M.; Marquez, R. Angew. Chem. Int. Ed. 2001, 40, 603. c) Evans, D.A.;
Ripin, D.H.B.; Halstead, D.P.; Campos, K.R. J. Am. Chem. Soc. 1999, 121, 6816. d) Evans, D.A.;
Carter, H.P.; Carreira, E.M.; Charette, A.B.; Prunet, J.A.; Lautens, M. J. Am. Chem. Soc. 1999, 121,
7540.
3) a) Soriente, A.; De Rosa, M.; Villano, R.; Scettri, A. Curr. Org. Chem. 2004, 8, 993. b) Villano, R.;
Acocella, M. R.; Massa, A.; Palombi, L.; Scettri, A. Tetrahedron Asymmetry 2006, 17, 3332. c)
Villano, R.; Acocella, M. R.; Massa, A.; Palombi, L.; Scettri, A. Tetrahedron 2009, 65, 5571. d)
Villano,R.; Acocella, M. R.; De Sio, V.; Scettri, A. Cent. Eur. J. Chem. 2010, 8, 1172. e) Villano, R.;
Acocella, M. R.; Scettri, A. Tetrahedron 2011, 67, 2768.
Oral 7
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
ASSEMBLY OF A SUCCINYL-CALIX[4]ARENE DERIVATIVE IN
MULTILAMELLAR VESICLES
Granata G.1, Consoli G. M. L.1, Geraci C.1, Lo Nigro R.2, Malandrino G.3
1
Istituto di Chimica Biomolecolare-C.N.R., Via P. Gaifami 18, 95126, Catania, Italy.
2
Istituto per la Microelettronica e Microsistemi-C.N.R., Strada Ottava 5, 95121,
Catania, Italy.
3
Dipartimento di Scienze Chimiche, Università di Catania, Viale A. Doria 6, 95125,
Catania, Italy.
email: [email protected]
Self-assembled structures built from amphiphilic molecules find application in
different fields. a
The synthetic versatility of calixarene macrocycles, whose skeleton can be
differently functionalized with a variety of hydrophilic and hydrophobic moieties,
allows the development of amphiphilic derivatives able to aggregate in different
higher order supramolecular architectures. b, c
Here we describe the capability of an amphiphilic calix[4]arene derivative, bearing
four carboxylic acid groups at the calixarene upper rim and four dodecyl chains at
the lower rim, to form discrete and stable multilamellar vesicles in aqueous
medium by using an emulsion/solvent evaporation method.
Dynamic Light Scattering (DLS) measurements, Field Emission Scanning Electron
Microscopy (FE-SEM) and Transmission Electron Microscopy (TEM) images
provided information about size, size distribution and morphology of the
nanostructured assemblies. The vesicular nature was confirmed by entrapment of
a hydrophilic dye such as rodhamine B in aqueous lumen.
A preliminary study showed that the self-assembled structures of the carboxycalix[4]arene can entrap also a hydrophobic drug such as curcumin.
The amphiphilic carboxy-calix[4]arene represents the first example of calix[4]arene
derivative assembling in very stable multilamellar vesicles in aqueous medium with
a narrow polydispersity. Suitable nanometer-size, very size narrow distribution,
well-defined spherical shape, negatively charged hydrophilic multilamellar wall,
high size stability (more than two years), and guest encapsulation ability make the
vesicles of the carboxy-calix[4]arene derivative appealing in the research of drug
delivery systems.
Acknowledgment: Research funded by PON R&C 02_00355_2964193 (MIUR, Rome).
References: a) Hill, J. P.; Shrestha, L. K.; Ishihara, S.; Ji, Q.; Ariga, K. Molecules 2014, 19, 85898609. b) Helttunen, K.; Shahgaldian, P. New J. Chem. 2010, 34, 2704–2714. c) Consoli, G. M. L.;
Granata, G.; Lo Nigro, R.; Malandrino, G.; Geraci, C. Langmuir 2008, 24, 6194–6200.
Oral 8
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
A ‘SEA’ OF BIOACTIVE NATURAL PRODUCTS
Cutignano A.
Institute of Biomolecular Chemistry – CNR, via Campi Flegrei 34 - 80078
Pozzuoli (NA)
email: [email protected]
The marine environment is distinguished by unique groups of organisms that hide
a treasure of fascinating and peculiar structures. The extraordinary chemical
diversity of marine organisms often mirrors potent biological activities that
represent the basis for the ecological function of marine natural products but also
a promise for pharmacological applications.
In our ongoing search for novel marine bioactive molecules we recently focused
our attention on species from less explored habitats and on cultivable microorganisms, either from commercial sources or isolated from the Mediterranean
Area.
Promising stories of bioactive natural or naturally inspired molecules, identified
following traditional approaches will be discussed, highlighting the results obtained
very recently through a multidisciplinary strategy that embraces expertise in the
fields of the natural product chemistry, organic synthesis and pharmacological
assays.
Furthermore, in the frame of two PON projects aiming at discover new
immunomodulant and anti-infective agents from marine sources, we validated and
applied an automated chromatographic platform to the screening of a library of raw
extracts from marine microalgae and sponges. The process is well suited for high
throughput screening program and entails dramatic time saving along with
effective desalting and reproducible fractionation of marine extracts. The potential
of the developed protocol will be illustrated through key examples of bioactive
molecules identified in few steps.
Acknowlegments: Research funded by PON Projects 01/00117 and 01/02093 and by PNRA
Project 2009/A1.06
Oral 9
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
NMR METABOLOMIC APPLICATIONS ON OLIVE OILS:
PEDOCLIMATIC AND SENSORY DIFFERENTIATION. FROM
SINGLE PULSE NMR TO ADIABATIC SEQUENCES
Culeddu N.1, Chessa M.1, Molinu M.G.2, Deiana P.3, Dettori S.3 and
Santona M.3
1) CNR – Institute of Biomolecular Biochemistry UOS Sassari, via La crucca
3, I-7100 Sassari
email: [email protected]
2) CNR – ISPA Institute Of Sciences of Food Production - UOS Sassari via La
crucca 3, I-7100 Sassari
3) University of Sassari - Dept. of Sciences for Nature and Environmental
Resources, via De Nicola 9 I-7100 Sassari.
Olive oil is a fundamental constituent of the traditional Mediterranean diet, which
is continuously attracting the interest of the scientific community for its healthprotecting activities (1). The protective role of olive oil in fighting certain diseases
has been attributed to its fatty acid composition and the presence of minor
constituents, mainly phenolic compounds and squalene. Properly extracted from
fresh ripe fruits, the oil is characteristically aromatic and slightly bitter. The
sensory characteristic of virgin olive oil has been related with the presence of
phenolic compounds derived from the hydrolysis of oleuropein, aldehydes and
terpenes, (2)
We show some models obtained by metabolomic analysis of NMR data obtained
from standard NMR sequences, multi suppression sequences and novel
selective excitation scheme to analyze the aldehydes and polyphenols in olive oil
without the need for extraction/concentration of these analytes.
These models take in account the relationship between NMR data vs
pedoclimatic description, soil composition and sensory analysis.
We recognized also the instability of oleuropein when it react with methanol,
leading to the artificial formation of isomers.
1) Frankel, E. N. J. Agric. Food Chem. 2011, 59, 785-792.
2) Boskou D., OLIVE OIL:Minor Constituents and Health CRC press 2009
Oral 10
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
ONE-POT MICROWAVE ASSISTED CATALYTIC TRANSFORMATION
OF WASTE FRYING OIL INTO GLYCEROL-FREE BIODIESEL
Drago C., Nicolosi G.
Istituto di Chimica Biomolecolare, CNR, via Paolo Gaifami 18, I-95126 Catania; Italy,
email: [email protected]
Biofuel production has increased in recent years and some commodities like corn,
sugar cane or vegetable oil can be used either as food, feed, or to make biofuels. An
alternative to overcome the "Food versus Fuel" dilemma is to use only non-food crops
as feedstock for biofuel production.a However, the massive exploitation of land for
biofuels production is often a consequence of its sterilization and desertification. So,
long-term sustainability for biofuels can be accomplished by exploiting alternative
feedstocks from non-arable land. A priceless and non-negligible feedstock would arise
from waste recyclability and their reuse. In this contest our attention is focused to the
WFOs (waste frying oils) from household and industrial sources.b Recently, We have
reported a catalytic one-pot microwave assisted transformation of vegetable oils to a
mixture of FAMEs (methyl esters of fatty acids) and glycerol ether derivatives, using
the commercially available TBME (tert-butyl metyl ether) as single source both as
transesterification and transetherification reagent.c,d The present study is an attempt to
adapt the already developed protocol to
R
the transformation of WFO, which we
O
O
O
R
R
O
received from a local restaurant, to a
O
O
final potential biofuel. It is necessary to
O
note that the high temperature of the
O
O
MW: power/time
MW: power/time
)))
)))
frying processes and the water from the
+
+
O
foods accelerate the hydrolysis of
CH OH
O
3
triglycerides, and it increases the FFAs
R
O
acid-catalyst
(Free Fatty Acids) content in the oil.
Usually, vegetable oils with high content of FFA are not preferred for biodiesel
production because they need a pretreatment to convert the FFA to the corresponding
methyl esters.e Preliminary analysis on the received WFO showed a quantity of FFAs
10 times major to the same oil before its use. When a sample of WFO and TBME
(1:10), in presence of 1% of the selected catalyst (w/w), was submitted to the
microwave radiation, in 3 hours, we observed a complete conversion into the
corresponding mixture of the corresponding FAMEs and a mixture of glycerol ethers
derivatives. Interesting to highlight a complete conversion of FFAs to FAMEs and, as
desired, a negligible traces of glycerol were detected.f There are several practical and
cost-saving advantages making this process very attractive, among which the WFO
could be directly transformed without any preliminary pretreatment, the obtained
mixture, which is without glycerol, only needs to be separated from the catalyst,
achieved by a very simple filtration, and it may be used directly in the energy chain.
3
Acknowledgements: The authors acknowledge the financial support provided by EFOR-CNR Project.
References: a) Atabani, A. E.; Silitonga, A. S.; et al.; Renew. Sust. Energ. Rev.; 2013, 18, 211-245. b)
Wermelinger, V. K.; Araujo; et al.; Bioresource Technology 2010, 101, 4415-4422. c) Drago, C.; Liotta,
L. F.; La Parola, V.; Testa, M. L.; Nicolosi, G.; PCT N° PCT/IB2014/058783. d) Drago, C.; Liotta, L. F.;
La Parola, V.; Testa, M. L.; Nicolosi, G.; Fuel, 2013, 113, 707. e) Melero, J. A.; Iglesias, J.; Morales, G.;
Green Chem. 2009, 11,1285–1308. f) Drago, C.; Nicolosi, G.; et al.; manuscript in preparation.
Oral 11
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
CARRYING ON THE ROUTE OF MARINE GUANIDINES:
ISOLATION, SYNTHESIS AND BIOACTIVITY OF A GUANIDINE
TERPENE FROM THE MOLLUSK ACTINOCYCLUS PAPILLATUS
Gavagnin M.1, Carbone M.1, Ciavatta M. L.1, Kiss R.2,
Mathieu V. 2, Mollo E.1, Guo Y.-W.3
1
Istituto di Chimica Biomolecolare, CNR, Via Campi Flegrei 34, 80078, Pozzuoli
2
Laboratoire de Cancérologie et de Toxicologie Expérimentale, Faculté de
Pharmacie, Université Libre de Bruxelles (ULB), Brussels, Belgium,
3
State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica,
CAS, Zu Chong Zhi Road 555, 201203,Shanghai
email: [email protected]
Guanidine is one of the most widely used scaffold in the design and synthesis of
new bioactive compounds. Numerous guanidine compounds from natural sources
as well as those obtained through synthetic strategies and virtual design have
gained much interest over the last few decades due to their therapeutic potential.ac
In nature the Y-shaped CN3 unit is mainly incorporated in peptide, polyketide,
and aromatic structures whereas only a limited number of guanidine compounds
possess a terpene framework. Despite their low occurrence, terpene guanidines
have shown interesting biological activities, standing out as promising lead
structures suitable for development of potential drugs. Prenylated guanidines from
higher plants, for example, display antitumor, anti-angiogenic, antimicrobial,
insulin-like and anti-lipolytic properties.c
Continuing our studies on guanidines from marine mollusks that have led in recent
years to the characterization of bioactive phidianidines,d-f we have now isolated
actinofide (1), from the Chinese nudibranch mollusk Actinocyclus papillatus.
The structure exhibiting two terpenoid acyl residues linked to a guanidine unit has
been determined by spectroscopic methods and confirmed by synthesis.
Actinofide (1) has been tested for the inhibition of cellular growth on a panel of
cancer cell lines showing activity at a concentration less than 10 M against
selected cell lines. Starting from the synthesis of 1, a small library of linear terpene
acyl guanidines (with C5, C15 and C20 fragments) have been also prepared for
the pharmacological screening.
References: (a) Rauf, M.K.; Din, I.; Badshah A. Expert Opin. Drug. Discov. 2014, 9, 39-53. (b)
Saczewski, F.; Balewski, L. Expert Opin. Ther. Patents 2013, 23, 965-995. (c) Berlinck, R.G.S.;
Trindade-Silva, A.E.; Santos M.F.C. Nat. Prod. Rep. 2012, 29, 1382-1406. (d) Carbone, M.; Li, Y.;
Irace, C.; Mollo, E.; Castelluccio, F.; Di Pascale, A.; Cimino, G.; Santamaria, R.; Guo, Y.-W.;
Gavagnin, M. Org. Letters 2011, 13, 2516–2519. (e) Manzo, E.; Pagano, D.; Ciavatta, M.L.;
Carbone, M.; Gavagnin M. 2013, PCT/IB2013/052,163; 2015, U.S. Patent 9.085.569. (f) Vitale,
R.M.; Gatti, M.; Carbone, M.; Barbieri, F.; Felicità, V.; Gavagnin, M.; Florio, T.; Amodeo, P. ACS
Chem. Biol. 2013, 8, 2762–2770.
Oral 12
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
Poster Presentations
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
INFLUENZA VIRUS PA ENDONUCLEASE: VIRTUAL SCREENING,
DOCKING AND IN-VITRO VALIDATION OF NOVEL INHIBITORS
Pala N.1, Dallocchio R.2, Dessì A.2, Cadoni R.1, Carcelli M.3, Stevaert
A.4, Sechi M.1, Naesens L.4
1
di Sassari, Via Vienna 2, 07100
Sassari, Italy
Istituto di Chimica Biomolecolare, CNR−Consiglio Nazionale delle Ricerche,
Sassari, 07100 Li Punti Italy, mail to [email protected], [email protected],
3
, Parco Area delle Scienze 17/A,
43124 Parma, Italy
4
Rega Institute for Medical Research, KU Leuven, Minderbroedersstraat 10, B3000 Leuven, Belgium
2
Vaccination and antiviral agents are the primary measures against human
influenza viruses. However, both interventions have shortcomings, and hence,
there is an urgent need to develop new and more efficacious anti-influenza virus
drugs. The influenza virus RNA-dependent RNA polymerase complex (RdRp), a
heterotrimeric protein complex that is responsible for viral RNA transcription,
represents an unexplored target for antiviral drug development. One particularly
attractive approach is interference with the endonucleolytic “cap-snatching”
reaction by the PA subunit of the RdRp, more precisely by inhibiting the metaldependent catalytic activity which resides in the N-terminal part of PA (PA-Nter).
In the last two decades, several small molecule PA inhibitors (PAIs) have been
discovered. Among them, compounds belonging to the class of substituted 2,4dioxobutanoic acids were identified as particularly potent and selective PAIs in
both enzyme and cell-based assays. A few other classes of potential PAIs have
been identified. All these diverse compounds bear distinct pharmacophoric
fragments with chelating motifs able to bind the bivalent metal ions in the catalytic
core of PA-Nter. More recently, the availability of crystallographic structures of
PA-Nter has enabled rational design of PAIs with improved binding properties.
Here, we present a new coupled pharmacophore/docking virtual screening
approach that allowed us to identify novel PAIs having interesting inhibitory activity
in a PA-Nter enzymatic assay, as well as antiviral activity in a cell-based influenza
virus yield assay.
References:
1) Stevaert, A.; Dallocchio, R.; Dessì, A.; Pala, N.; Rogolino, D.; Sechi, M.; Naesens, L.; J. Virol.
2013, 87, 10524−10538. 2) Pala, N.; Dallocchio, R.; Dessì, A.; Brancale, A.; Carta, F.; Ihm, S.;
Maresca, A.; Sechi, M.; Supuran, C. T.; Bioorg. Med. Chem. Lett. 2011, 21, 2515−2520. 3) Pala, N.;
Stevaert, A.; Dallocchio, R.; Dessì, A.; Rogolino, D.; Carcelli,M.; Sanna, V.; Sechi, M.; Naesens, L.;
ACS Med. Chem. Lett. 2015, Just Accepted Manuscript DOI: 10.1021/acsmedchemlett.5b00109.
Poster 1
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
THE HEMORPHINS: A NEW CLASS OF OPIOID PEPTIDES
DERIVED FROM HEMOGLOBIN
Fenude E.1, Vite V.2
1
ICB, Trav. La Crucca n.3, 07040, Sassari
Corso di Laurea Specialistica in Chimica e Tecnologie Farmacautiche,
Dipartimento di Chimica e Farmacia, Università di Sassari
email: [email protected]
2
Many biologically active peptides are generated by proteolytic processing of
various higher molecular weight multifunctional precursor peptides and proteins. A
series of ”nonclassical” endogenous opioid peptides (hemorphins) have been
identified in the course of the study of proteolytic fragments of bovine blood
hemoglobin. Depending on the N-terminal sequences the sub-families of LVVemorphins, VV-hemorphins, and V-hemorphins can be distinguished. It was shown
that the overall activity varied considerably among different hemorphins which
points to the importance of the N-terminal hydrophobic residues on structurefunction relationship. It is well accepted that the binding of the ligand to its receptor
is mediated by ion-ion interactions, hydrogen bonding, dipole-dipole interactions,
lipophilicity, and shape complementarity but the relative contributions of each of
these interactions is still poor understood. We perused this goal by examination of
the conformational and dynamic properties of synthetic analogues sequences
carefully selected for their complementary biological properties. In this work it has
been observed tetra- and penta-peptides fragments of N-terminal protected LVVhemorphins which are well known to be opioid receptor ligands and
immunoregulatory peptides in order to study the structural-functional peculiarity.
Structural study in CHCl3 has been directed towards peptides with the follow
aminoacidic sequence: Boc-Leu-Val-Val-OMe, Boc-Leu-Val-Val-Tyr-OMe, Boc-LeuVal-Val-Phe-OMe, Boc-Leu-Val-Val-Tyr-Pro-OMe. These products have been
synthesized, purified, and then analyzed by NMR spectroscopy, employing both
mono- and bi- dimensional homo- and hetero- nuclear correlation 1H-1H, 1H-13C
techniques through which it is possible to obtain structural and conformational
informations. The result obtained are compared and discussed.
References
Fenude E., Dettori A., Demontis M.P., Alberico E., & al., Pharm. Res., 2003, 47, 53
Fenude E., Dedola S., Fais M., VII Convegno “Complex Systems: structure,
properties, reactivity and dynamics, 2005, Alghero, 13-15 Giugno
Fenude E., Roggio A.M., XXII Congresso Nazionale della Società Chimica
Italiana, 2006,10-15 Settembre Firenze
Poster 2
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
POLYSACCHARIDE-BASED CHIRAL STATIONARY PHASES:
EXPLOITING THE SIZE AND SHAPE OF THE MOLECULES FOR
CHIRAL RECOGNITION AND ENANTIOSEPARATIONS
Peluso P.
Istituto di Chimica Biomolecolare, UOS di Sassari
Traversa La Crucca, 3 – Regione Baldinca, Li Punti, 07100 Sassari
email: [email protected]
In keeping with the importance of chirality, chemistry invests resources to access
pure enantiomers, reproducing homochirality in xenobiotic chemical contexts and
understanding the mechanisms that govern the chiral discrimination processes.
The real meaning of chirality lies in the relational and directional sense of the
word. It means that two enantiomers have exactly the same properties in isotropic
conditions. On the contrary, a chiral enantiopure molecule can recognize two
enantiomers through stereoselective and anisotropic interactions. Nowadays,
chemists largely use chromatography on chiral support for resolving racemic
mixtures and a number of reasons make HPLC the preferred technique for
enantioseparations. In the last decades, polysaccharide-based chiral stationary
phases (CSPs) have been preferred for HPLC enantioseparations and, currently,
the market makes different chiral columns available, which contain selectors based
on cellulose and amylose derivatives. In this field, understanding the chiral
recognition mechanisms is still a challenge.
The results obtained in this study a) prove that the anisotropic properties of the
molecules as well as their size and shape deeply influence the enantioseparability a
and b) explain why similar molecules produce very different enantioseparations (A)
or why very diverse molecules are able to give analogous separations (B).b-d
Acknowledgment: Research funded by Università Ca’ Foscari di Venezia (ADIR funds) and
th
Regione Autonoma della Sardegna (L.R., August 7 2007, n. 7, call 2008).
References: a) Peluso, P.; Cossu, S. Chirality 2013, 25, 709-718. b) Peluso, P.; Mamane, V.;
Aubert, E.; Cossu, S. J. Chromatogr. A 2012, 1251, 91-100. c) Peluso, P.; Mamane, V.; Aubert, E.;
Cossu, S. J. Chromatogr. A 2014, 1345, 182-192. d) Peluso, P.; Mamane, V.; Cossu, S. Chirality
2015, DOI: 10.1002/chir.22485.
Poster 3
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
NOVOZYM-435 AS EFFICIENT CATALYST FOR THE SYNTHESIS
OF BENZOIC AND (HETERO)AROMATIC CARBOXYLIC ACID
ESTERS
Sechi B., Giunta D., Marchetti M., Solinas M.
Istituto di Chimica Biomolecolare, UOS di Sassari
Traversa La Crucca, 3 – Regione Baldinca, Li Punti, 07100 Sassari
email: [email protected]
Hereby we report new reaction conditions to convert benzoic acids and more
general (hetero)aromatic carboxylic acids into the corresponding n-heptyl esters
by applying Novozym-435 as the biocatalyst in cyclohexane as the solvent. Very
good yields are obtained in the esterification of a plethora of substituted acids with
mild and straightforward reaction conditions. Direct esterification of the acid is
favoured compared to transesterification of methyl benzoate under our reaction
conditions in all cases studied. Recycling of the immobilised enzyme is feasible
although with some minor limitations.
th
Acknowlegment: Research funded by Regione Autonoma della Sardegna (L.R., August 7 2007, n.
7, CRP_25257)
References: a) Daniela Giunta, Maria Paola Masia, Mauro Marchetti, Raffaele Morrone, Maurizio
Solinas Tetrahedron Letters 54 (2013) 5122–5125 b) Daniela Giunta, Barbara Sechi, Maurizio
Solinas Tetrahedron 71 (2015) 2692-2697.
Poster 4
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
EFFICIENT BASE-FREE HYDROGENATION OF AMIDES TO
ALCOHOLS AND AMINES CATALYSED BY WELL-DEFINED
PINCER IMIDAZOLYL-RUTHENIUM COMPLEXES
Alberico E.1, Cabrero-Antonino J. R.2, Drexler H.-J.2, Baumann W.2,
Junge K.2, Junge H.2, Beller M.2
1
Istituto di Chimica Biomolecolare, Consiglio Nazionale delle Ricerche,
Tr. La Crucca 3, 07100 Sassari (Italy); email: [email protected]
2
Leibniz-Institut für Katalyse e.V. Albert Einstein Str. 29a, 18059 Rostock
(Germany).
Among the methods amenable to the synthesis of amines, catalytic hydrogenation
of amides using molecular hydrogen1 is the most promising as to atom economy
and waste prevention when compared to the use of LiAlH4 and B2H6 or metal
promoted catalytic hydrosilylation. In order to overcome the low electrophilicity of
their carbonyl groups the reduction of amides with molecular hydrogen is usually
carried out at elevated pressures and temperatures, although recent improvement
of heterogeneous catalysts has witnessed a mitigation of reaction conditions. The
main drawback associated with the use of such catalysts is that they are
incompatible with aromatic groups and double/triple CC bonds which are likewise
reduced. This problem has been recently overcome by the development of a
homogeneous
ruthenium
catalyst
modified
by
1,1,1tris(diphenylphosphinomethyl)ethane (Triphos) which operates in the
presence of an acid cocatalyst. Yet it has a rather limited scope being best suited
for 1° amides or substrates which bear a phenyl ring directly attached to the
nitrogen atom and requires temperatures above 200 °C.
Heterogeneous catalysts and the Ru/Triphos system promote the reduction of
amides according to path a of Scheme 1. Homogeneous catalysts which rely on
metal-ligand cooperation (bifunctional catalysis) instead pave the way to an
alternative reactivity (Scheme 1, path b) in which the initial reduction of the
carboxylic group is followed by collapse of the intermediate hemiaminal by
cleavage of the C-N bond and reduction of the resulting aldehyde, overall affording
the alcohol and amine.
Herein we report the synthesis of two
novel ruthenium pincer complexes bearing
an imidazolylaminophosphino pincer
ligand.2 The BH4-substituted one has
demonstrated to be an efficient catalysts
for the hydrogenation of different
substituted amides to the corresponding
alcohols and amines in excellent yields
without the use of additional base.
Scheme
Acknowlegment: Research funded by the ACS Pharmaceutical Roundtable
References: 1) Smith A. M., Whyman R., Chem. Rev. 2014, 114, 5477; 2) Cabrero-Antonino J. R.
,Alberico E., Drexler H.-J., Baumann W., Junge K., Junge H., Beller M. submitted to Chem.
Commun.
Poster 5
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
MOLECULAR DATABASE IMPLEMENTATION
Cosseddu A.
Institute of Biomolecular Chemistry (ICB), Li Punti, 07100 Sassari
email: [email protected]
Implementation of Molecular Database ICB Support Units (UOS) located in Sassari has
begun as activity in several projects to which the UOS of Sassari has participated. The
database has been then extended and integrated with the largest and most
comprehensive molecular database "StOrMoDB" generated by ICB (Amodeo P. et al).
Over a span of time from 2000 to the present, new 915 molecules synthetized from five
research groups of UOS of Sassari have been classified.
The classification of individual molecules has been made through a common pattern that
describes the main features such as structure, identification, purification, solubility,
characterization (by NMR spectra, melting points, IR spectra, etc.), any biological activity.
Each hit reports authors and scientific papers by which molecules has been described.
Structure
Identification
Key Words: alfaSynuclein, Biphenyl
compounds,
Curcumin-like
molecules, Parkinson’s
disease,
Protein aggregation
Label: VD040
Chemical Name:
(3E,3'E)-4,4'-(6,6'-dihydroxy-5,5'dimethoxy-[1,1'-biphenyl]-3,3'diyl)bis(but-3-en-2-one)
Molecular Formula: C22H22O6
Molecular Weight: 382.41
Melting Point:
242-243 °C
Source:
synthetic compound
Solubility:
soluble in most
common organic
solvents
Purification: filtration
Spectra
1H, 13C
NMR
Bioattivity/Ecotox
Effect:
-Synuclein interaction
-Synuclein aggregation
Test-System:
circular dichroism and
fluorescence spectroscopy
Antioxidant activity
Test-System: DPPH
assays
Test-System: TEAC
assays
Neuroprotective
activity
Test-System:
Cell viability
Species:
rat pheochromocytoma
(PC12) cells
Reference
Amino Acids, 2013, 45:327–338
DOI:
10.1007/s00726-013-1503-3
CNR _ICB_UOS Padova
[email protected]
CNR _ICB_UOS Sassari
[email protected]
[email protected]
[email protected]
Food Chemistry, 2014, (157),
263–274
DOI:
10.1016/j.foodchem.2014.02.036
Institute of Organic
Chemistry, Bulgarian
Academy of Sciences
[email protected]
CNR _ICB_UOS Sassari
[email protected]
[email protected]
[email protected]
In addition to entering into the most comprehensive Molecular Database StOrMoDB further
application will be the possibility to carry out studies of the structure-activity against
biological targets.
Acknowlegment: Research funded by
BioTTasa - MISE- Dipartimento per lo Sviluppo e la Coesione Economica - Bando RIDITT
Salude e Trigu – Regione Autonoma della Sardegna (L.R., August 7th 2007, n.7)
Cluster Chimica Verde – MIUR - Avviso D.D. n. 257/Ric del 30 maggio 2012
Poster 6
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
ORGANOCATALYTIC, ASYMMETRIC ELIMINATIVE [4+2]
CYCLOADDITION OF ALLYLIDENE MALONONITRILES WITH
ENALS: RAPID ENTRY TO CYCLOHEXADIENE EMBEDDING
LINEAR AND ANGULAR POLYCYCLES
Zambrano V.1, Rassu G.1, Pinna L.2, Brindani N.3, Dell’Amico L.3,4,
Curti C.3, Sartori A.3, Battistini L.3, Casiraghi G.3, Pelosi G.5, Greco D.3,
Zanardi F.3
1 Istituto di Chimica Biomolecolare, Consiglio Nazionale delle Ricerche, Li Punti
Sassari, Italy, 2 Dipartimento di Chimica e Farmacia Università degli Studi di
Sassari, Sassari, Italy, 3 Dipartimento di Farmacia,Università degli Studi di Parma,
Parma, Italy. 4 Institute of Chemical Research of Catalonia, Spain Tarragona,
5 Dipartimento di Chimica, Università degli Studi di Parma, Parma, Italy
email: [email protected]
Six-membered carbocycles, as well as five- and seven membered rings, are
common motifs in nature and they can be found in many terpenoid, polyketide,
and shikimate derived monocyclic and polycyclic molecular architectures. 1
A direct aminocatalytic synthesis has been developed for the chemo-, regio-,
diastereo-, and enantioselective construction of densely substituted polycyclic
carbaldehydes containing fused cyclohexadiene rings2. The chemistry utilizes, for
the first time, remotely enolizable -extended allylidenemalononitriles as electronrich 1,3-diene precursors in a direct eliminative [4+2] cycloaddition with both
aromatic and aliphatic -unsaturated aldehydes. The generality of the process is
demonstrated by approaching 6,6-, 5,6-, 7,6-, 6,6,6-, and 6,5,6-fused ring systems,
as well as biorelevant steroid-like 6,6,6,6,5- and 6,6,6,5,6-rings.
A stepwise reaction mechanism for the key [4+2] addition is proposed as a domino
bisvinylogous Michael/Michael/retro-Michael reaction cascade. The utility of the
malononitrile moiety as traceless activating group of the dicyano nucleophilic
substrates is demonstrated.
1) P. M. Dewick in Medicinal Natural Products. A Biosynthetic Approach, 2nd ed., Wiley, Chichester,
2002. 2) N. Brindani, G. Rassu, L. Dell’Amico, V. Zambrano, L. Pinna, C. Curti, A. Sartori, L.
Battistini, G. Casiraghi, G. Pelosi, D. Greco, F. Zanardi Angew. Chem. Int. Ed. 2015, 54, 7386 –
7390.
Financial support provided by the Università degli Studi di Parma
Poster 7
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
DETERMINATION OF FLAVOUR AND PIGMENT PROFILES IN
PRE- AND POST-STABILIZATION FRUIT-BASED FOOD
FORMULATIONS
Sperlinga E., Napoli E., Siracusa L., Ruberto G.
ICB – CNR UOS CATANIA, Via Paolo Gaifami,18, 95126 Catania
email: [email protected]
Shelf-life is defined as the maximum recommended time for which foods can be
stored before becoming unsafe for consumption.a) In order to evaluate the shelflife of a given food product there are many important parameters to consider, such
as those related to its organoleptic features. The present research activity was
carried out in collaboration with Dolfin s.p.a., a confectionery factory located in the
province of Catania which provided the fruit-based formulation object of the study.
Five aromatic commercial formulations (lemon, orange, berries, black cherry,
strawberry) and three pigment commercial formulation (grape, shade lemon
yellow, shade apple green) were used to proper set the analytical methods. The
fruit-based formulation (apple mash, strawberry juice, water, strawberry flavour,
natural red pigment) was subjected to accelerated shelf-life tests at different
storage times (T0, T15, T30, T48, T62). Stabilization tests at room temperature
were carried out after 90 and 180 days of storage. Finally, 3 different kind of
secondary packaging were tested (984, NKME, cardboard) and they were
compared to the one that is actually used. All the samples were subjected to the
classical pasteurization process. Aroma compounds were extracted using the solid
phase micro extraction (SPME) technique and the determination was carried out
through GC-MS analysis. Pigments and phenolics were extracted using a solution
of methanol/water/hydrochloric acid (80:19:1) and the extracts analyzed by HPLCDAD, focusing on anthocyanins & cinnamic acids.
Results for accelerated tests samples showed that, after 15 days of storage,
there was a significant (80%) decrement in anthocyanin content, which fell to more
than 96% after 30 days. Chlorogenic acid (the dominant phenol in the formulation)
was subjected to a slower degradation process; in fact, its content decreased of
less than 50% after 62 days. Regarding flavours (mainly methyl cinnamate, ethyl
cinnamate and -decalactone) a quite stable trend all throughout the shelf-life tests
was observed. Among the minor components, hexenyl acetate, ethyl eptanoate
and α-terpineol were the ones that decreased more, while acetophenone and butyl
isovalerate rose up. After 15 days, benzaldehyde, hexyl acetate and ethyl
octanoate were formed. There were no significant differences among the other
samples.
Acknowledgment: Research funded by “Programma Operativo Nazionale Ricerca e Competitività
(R&C)”, 2007‐2013:SHELF‐LIFE PON02_00451_336190.
References: a) Pasha, I.; Saeed, F.; Sultan, M.T.; Khan, M.R.; Rohi, M. Crit. Rev. Food Sci. Nutr.
2014, 54, 340-351.
Poster 8
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
EFFECTS OF TREHALOSE ON ALPHA-SYNUCLEIN STABILITY
REVEALED THROUGH SYNCHROTRON RADIATION CIRCULAR
DICHROISM
Ruzza P.1, Hussain R.2, Biondi B.1, Calderan A.1, Tessari I.3,
Bubacco L.3 and Siligardi G.2
1
Institute of Biomolecular Chemistry of CNR, Padua Unit, Padua 35131, Italy;
2
Diamond Light Source Ltd., Harwell Innovation Campus, Chilton, Didcot,
Oxfordshire OX11 0QX, UK;
3
Department of Biology, University of Padua, Padua 35122, Italy.
email: [email protected]
Protein misfolding and aggregation characterized many neurodegenerative
diseases, including Huntington’s, Alzheimer’s and Parkinson’s diseases. Recently,
several research groups found that trehalose and other osmolytes are able to
interfere with protein misfolding and aggregation [a-c].
In the present work, we studied, by means of synchrotron radiation circular
dichroism (SRCD) spectroscopy, the dose-effect of trehalose on α-synuclein
conformation and/or stability to probe the capability of this osmolyte to interfere
with α-synuclein’s aggregation.
Our results indicated that a low trehalose concentration stabilizes α-synuclein
folding much better than at high concentration by blocking in vitro α-synuclein’s
polymerization [d].
These results suggested that trehalose could be associated with other drugs
leading to a new approach for treating Parkinson’s and other brain-related
diseases.
Acknowlegment.
We thank Diamond Light Source for access to beamline B23 (SM8034 and SM9079) that
contributed to the results presented here. The research leading to these results has received
funding from the European Community’s Seventh Framework Programme (FP7/2007-2013) under
grant agreement nº 226716.
References.
a. Tanaka, M.; Machida, Y.; Niu, S.Y.; Ikeda, T.; Jana, N.R.; Doi, H.; Kurosawa, M.; Nekooki, M.;
Nukina, N. Nature Medicine 2004, 10, 148-154.
b. Liu, R.; Barkhordarian, H.; Emadi, S.; Park, C.B.; Sierks, M.R. Neurobiology of Disease 2005,
20, 74-81.
c. Jiang, T.; Yu, W.B.; Yao, T.; Zhi, X.L.; Pan, L.F.; Wang, J.; Zhou, P. RSC Adv. 2013, 3, 9500-9508.
d. Ruzza, P.; Hussain, R.; Biondi, B.; Calderan, A.; Tessari, I.; Bubacco, L.; Siligardi, G.
Biomolecules 2015, 5, 724-734
Poster 9
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
IN SILICO IDENTIFICATION AND EXPERIMENTAL VALIDATION OF
NOVEL MULTI-TARGETED ANTI-ALZHEIMER’S LIGANDS FROM MARINE
SOURCES
Vitale R.M.1, Desiderio D.4, Sgammato R.4, Thellung S.3, Carbone M.1,
Ciavatta M.L.1, Mollo E.1, Gavagnin M.1, Felicità V.1,2, Rispoli V.2, Florio T.3,
Masullo M.4, Amodeo P1
1 Instituto di Chimica Biomolecolare (ICB)-CNR, Via Campi Flegrei 34, 80078 Pozzuoli.
email: [email protected]
2.Dipartimento di Scienze della Salute, Università Magna Græcia of Catanzaro, Campus
“Salvatore Venuta”, Viale Europa,I-88100 Catanzaro (CZ), Italy
3 Sezione di Farmacologia, Dipartimento di Medicina Interna (DiMI), e Centro di
Eccellenza per la Ricerca Biomedica (CEBR) Università di Genova, viale Benedetto XV, 2,
16132 Genova
4 Dipartimento di Scienze Motorie e del Benessere Università degli Studi di Napoli
"Parthenope"
Alzheimer’s disease (AD), the most common cause of dementia in elderly population, is a
complex neurodegenerative disorder of the central nervous system, characterized by a
progressive and irreversible degeneration of cholinergic neurons with the concomitant
decrease of the hippocampal and cortical neurotransmitter acetylcholine (Ach) levels, and
by the presence of amyloid plaques and neurofibrillary tangles. The current
pharmacological therapy for AD is mainly based on acetylcholinesterase inhibitors
(AChEI), aimed at restoring the Ach tone in the brain. However, due to the multifactorial
aetiology of this pathology, compounds able to act simultaneously on two or more relevant
biological targets is high desirable, as combining AChE inhibition with the impairment of βamyloid peptide's aggregation and deposition by inhibition of β-secretase enzyme (BACE1) and/or by directly hampering Aβ aggregation.
The previous identification of a pseudozoanthoxanthin variant from the zoanthid crust coral
Parazoanthus axinellae, as an acetylcholinesterase inhibitora prompted us to search other
molecules sharing a similar scaffold in our collection of natural marine compounds, and to
explore the potentiality of the newly-selected molecules as multiligand agents, by including
as potential target BACE-1 and investigating the ability of this scaffold to inhibit β-amyloid
aggregation. The most similar compounds among those available in suitable quantities for
subsequent experimental validation within our collection were a pseudozoanthoxanthin
from an unidentified caribbean zoanthid, differing from the analog reported by Turk and
coll. for the number and position of methyl substituents on the azulene ring, and the
bromo-pyrrole alkaloid stevensine. Docking and molecular dynamics studies were carried
out on both AChE and BACE-1 enzymes as well as on β-amiloid fibrils. The positive results
obtained in silico were then confirmed by biochemical assays that showed an inhibition for
both enzymes in the low/sub-micromolar range as well as the ability of impairing β-amiloid
aggregation.
Acknowlegment: Research funded by POR Campania FESR 2007-2013 “FARMABIONET Rete integrata per
le
biotecnologie applicate a molecole ad attività farmacologica”
References: a) Sepcić K, Turk T, Macek P. Toxicon. 1998, 36:937-40.
Poster 10
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
CALIX[8]ARENE BASED PH-RESPONSIVE HYDROGELATORS:
USEFUL PLATFORM FOR DRUG DELIVERY
Cunsolo F., Mecca T.
CNR-Istituto di Chimica Biomolecolare, UOS of Catania,
Via P. Gaifami 18, 95126, Catania
email: [email protected]
Delivering drugs with improving safety and stability, controlled rate, slow delivery,
targeted delivery is the aim that researchers of this area pursue with very different
and attractive methods. Among the others, low molecular weight hydrogelators
with stimuli responsive functionalities are considered to be of great interest as drug
delivery systems.a Recently we have reported the first example of calix[8]arene
polyelectrolyte derivatives as pH-responsive hydrogelators able to form gel phases
in basic and acidic conditions.b In order to evaluate the utility of these hydrogels as
delivery systems, two different set of hydrogels were prepared. The first one,
obtained through basic pH-responsive calix[8]arene hydrogelators, has been
studied for the loading and release capacity towards steroid and non-steroid antiinflammatory drugs. Acidic pH-responsive calix[8]arene hydrogelators were
instead used to check the improvement of aqueous solubility and stability of
curcumin, a natural antioxidant that shows many pharmacological and protective
activities, for which many nanoformulations have been proposed in recent years. c
Acknowlegment: Research funded by PON R&C 02_00355_2964193 (MIUR, Rome).
References: a) Fleige, E.; Quadir, M. A.; Haag, R. Adv. Drug Delivery Rev. 2012, 64, 866-884.
b) Mecca, T.; Messina, G. M. L.; Marletta, G.; Cunsolo, F. Chem. Commun. 2013, 49, 2530-2532.
c) Naksuriya, O.; Okonogi, S.; Schiffelers, R. M.; Hennink, W. E. Biomaterials 2014, 35, 3365-3383.
Poster 11
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
PHOSPHOMANNOMUTASE 2: A NEW POTENTIAL TARGET FOR THE
THERAPY WITH PHARMACOLOGICAL CHAPERONES
Andreotti G.1, Cimmaruta C.1,2, Citro V.2, Cubellis M.V.2
1 Istituto di Chimica Biomolecolare – CNR, via Campi Flegrei 34, 80078 Pozzuoli-Napoli
email: [email protected]
2 Dipartimento di Biologia, Complesso Universitario di Monte S. Angelo,
Università degli Studi di Napoli Federico II,
via Cinthia, 21, 80126 NAPOLI
[email protected]
The most frequent glycosylation (CDG) disorder affecting the N-glycosylation pathway is
caused by a deficiency of Phosphomannomutase (PMM2) the enzyme that isomerize
Mannose-6-Phosphate. For this disorder, which is known as CDG-PMM2 (MIM: 212065)
[a], there is no therapy at present, but, at least in principle patients could benefit from a
therapy based on pharmacological chaperones (PC)[b]. PC are small molecules that
preferentially bind the folded state of a protein and stabilize it. Disease mutations can
lower the free energy difference between the folded and the unfolded protein shifting the
equilibrium towards the latter one. Unstable proteins, although retaining the functional
chemical groups needed for the biological activity, are sensitive to proteolysis and are
cleared by the protein quality control systems in the cell. Hence for these mutations the
reduction of the protein concentration in the cell is the primary effect and the reduction of
total activity is only a secondary effect.
Exploiting non-biased ligand migration and protein dynamics, we have produced the first
accurate model of a ligand bound complex for PMM2 which can be used for high
throughput screening to find PC for CDG-PMM2[c]. We have characterized in vitro the
mutants that are most commonly found in European population, we have demonstrated
that the genotype R141H/F119L is eligible for the therapy with PC. Moreover we have
demonstrated that a small molecule, glucose-1,6 bisphosphate acts as PC for F119L
stabilizing the mutant protein towards thermal induced denaturation and proteolysis. The
same metabolite enhances the activity and promotes the correct quaternary structure of
the mutant enzyme [d].
Acknowlegment: Research funded by Telethon (GGP12108)
References: a) J. Jaeken, Congenital disorders of glycosylation, in: M.L. O. Dulac, and H.B. Sarnat, Editors
(Ed.) Handbook of Clinical Neurology;Pediatric Neurology, vol. 113, Elsevier B.V, 2013, pp. 1737-1743. b)
H.H. Freeze, Towards a therapy for phosphomannomutase 2 deficiency, the defect in CDG-Ia patients,
Biochim Biophys Acta, 2009, 1792, 835-840. c) G. Andreotti, I. Cabeza de Vaca, A. Poziello, M.C. Monti, V.
Guallar, M.V. Cubellis, Conformational Response to Ligand Binding in Phosphomannomutase2: INSIGHTS
INTO INBORN GLYCOSYLATION DISORDER, J Biol Chem, 2014, 289, 34900-34910. d) G. Andreotti, E.
Pedone, A. Giordano, M.V. Cubellis, Biochemical phenotype of a common disease-causing mutation and a
possible therapeutic approach for the phosphomannomutase 2-associated disorder of glycosylation, Mol
Genet Genomic Med, 2013, 1, 32-44.
Poster 12
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
BIOTRANSFORMATION AND PREFERENTIAL CRYSTALLIZATION:
TWO PRACTICAL APPROACHES FOR THE RESOLUTION OF
MILNACIPRAN
Sanfilippo C. and Patti A.
1
Istituto di Chimica Biomolecolar-CNR, via P. Gaifami 18, 95126, Catania
email: [email protected]
Enzymatic catalysis is a valuable approach used in organic synthesis to achieve chemical
transformations in mild reaction conditions and with a high degree of selectivity. The use of
enzymes, especially lipase, in organic solvent is an advantageous way to obtain single
stereoisomers of a drug and biotransformations are today accepted as a powerful
methodology for the industrial preparation of chiral pharmaceuticals.
Enantiomerically pure molecules containing amino functions are important synthons for the
preparation of drugs. Primary and secondary amines, where the amino group is directly
located on a stereogenic carbon, have been obtained in good yields and optical purity by
lipases-catalyzed kinetic resolution of the racemates through a transamination reaction in
the presence of carboxylic esters as acyl donors.a However, aminomethyl compounds are
challenging substrates for their high reactivity and distance from the chiral center.
Milnacipran, Z-(±)-2-(aminomethyl)-N,N-diethyl-1-phenyilcyiclopropane, is an active
antidepressant drug belonging to the class of inhibitors of the reuptake of serotonin and
has recently attracted interest for its painkiller effects in the treatment of fibromyalgia.b
Milnacipran is currently marketed in many countries, but not yet in Italy, in racemic form,
however, recent pharmacokinetic studies on single enantiomers showed greater activity for
(1S, 2R)-levomilnacipran.
The aim of this study is the development of a simpler and more economical strategy
alternative to the reported an enantioselective synthesis. The kinetic resolution of racemic
milnacipran in the presence of lipase was then investigated and optimized by means of a
careful choice of the reaction conditions (lipase source and form, temperature, solvent and
acyl donor nature). The amide product and the unreacted substrate were obtained in
satisfactory chemical yields and enantiomeric purities.c
During this study we identified one of the amides obtained from the enzyme-catalyzed
resolution as a conglomerate, whose nature was confirmed by the physical properties
(melting points, solubility and X-ray diffraction). The properties of this specific amide of
milnacipran were then exploited for its spontaneous resolution by conglomerate
crystallization through a sequence of crystallization steps. Starting from a substrate with
quite low enantiomeric excess, crystals of both enantiomers were obtained in enantiopure
form.
References: a) Gotor-Fernández, V.; Busto, E.; Gotor, V.; Adv. Synth. Catal. 2006, 348, 797-812. b)
Ormseth, M. J.; Eyler, A. E.; Hammonds, C. L.; Boomershine, C. S.; J. Pain Res. 2010,3,15-24.c) Sanfilippo,
C.; Nicolosi, G.; Patti, A. J. Mol. Catal. B: Enzym. 2014, 104, 82-86
Poster 13
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
ANTIOXIDANT ACTIVITY MEASUREMENTS BY
ELECTRON SPIN RESONANCE SPECTROSCOPY
Sanna D.1, Fadda A.2, Serra M.1
1
2
Institute of Biomolecular Chemistry,
Institute of Sciences of Food Production, trav. La Crucca, 07040, Sassari
email: [email protected]
Antioxidant activity measurements are currently used in many fields ranging from
the cosmetic to the food industry. A variety of methods have been developed to
assess antioxidant activity (AA), but the complexity of substrates like food or
biological matrixes prevents from having a reliable method for antioxidant activity
quantification.a The preferred methods for AA determinations are those which
imply simple protocols, involve easy or none sample preparation, entail simple
instrumentation, like a spectrophotometer. These simple experimental procedures
allow for the determination of the AA of a lot of samples in a short time, but have
some disadvantages. The more easily predictable ones are interferences and
limitations of the method itself.
When considering the outcome provided, in AA measurements, by simple
spectrophotometric methods and when comparing these results with those
obtained with complementary and alternative methods we reached the conclusion
that those based on Electron Spin Resonance (ESR) spectroscopy are more
reliable.b In fact these latter involve the determination of relatively stable radical
species or the spin trapping of unstable radicals to form stable adducts. For this
reason, since only species with unpaired electrons are detected, the interferences
are scarce or missing. The comparison between the results obtained with
spectrophotometric and ESR determination of DPPH scavenging activity is a clear
example.
The absorbance of the oxidation products of the antioxidants and aggregation
phenomena which take place when using organic solvent/water mixtures can
prevent the use of the spectrophotometric method but are of no account when
using ESR spectroscopy.
Another example is the spectrophotometric determination of the superoxide radical
by using nitroblue tetrazolium. Attempts to generate this radical species, both with
enzymatic or chemical methods, and spin trapping experiments with DMPO or
DEMPO, were in most of the cases unsuccessful demonstrating that, in most of
the cases, this species is invoked but not really involved in these systems.
References:
a) Huang, D.; Ou B.; Pior, R.L. J Agric Food Chem 2005, 53,1841–1856
b) Sanna, D.; Delogu, G.; Mulas, M.; Schirra, M.; Fadda, A. Food Anal. Methods, 2012, 5, 759-766
Poster 14
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
BIOCATALYZED PROCEDURES TO RECOVER VALUABLE
PRODUCTS FROM FISH WASTE
Morrone R., Baglieri A., Gambera G., Nicolosi G., D’Antona N.
CNR – Institute of Biomolecular Chemistry UOS Catania, Via P. Gaifami 18 95126 Catania, Italy
email: [email protected]
The waste derived from the fish processing represents approximately 45-50% of
the weight of the raw material. Disposal of these wastes is expensive and
produces environmental issues, therefore their treatment as “novel” raw material
for the recovery of valuable products is certainly a valid alternative. Actually, the
main derivatives obtained from fishing wastes are fish meal and oil.
The oil from fish waste is commonly used to produce biodiesel but it represents
too an excellent source of long chain polyunsaturated fatty acids (PUFAs) and
especially of the precious omega-3 fatty acids, cis-5,8,11,14,17- eicosapentaenoic
acid (EPA) and cis-4,7,10,13,16,19-docosahexaenoic acid (DHA).
The functional and biological properties of omega-3 fatty acids include among
other: prevention of atherosclerosis, reduction in cardiovascular disease,
protection against arrhythmias, and reduction of the blood pressure a. At least two
meals of fish every week have been recommended by the American Heart
Association to reduce the effect of cardiovascular diseases b.
Since EPA and DHA are valuable products due to their beneficial properties on
human health, the demand for fish oil is constantly growing and so any new
sources are welcomed by the market.
The main problem in the fish oil extraction procedures is the lability (mainly due to
oxidative processes) of PUFAs when severe conditions, in terms of temperature
and pH, are requested. To avoid such harsh conditions different “softer” enzymatic
methods have been proposed c.
The following work is focused on the evaluation, at a laboratory scale, of different
proteases to recover oil from fish waste (head, bones, tail and gut).
The adopted mild reaction conditions allowed avoiding any degradation of PUFAs.
The fatty acid composition of the recovered oil was determined. Moreover, since
omega-3 are commonly formulated and marketed as their corresponding ethyl
esters too, an efficient irreversible biocatalyzed transesterification process in
absence of organic solvents was used.
Acknowlegment: Bio4Bio project, PON02_00451_3362376
References:
a) Kim S.K., Mendis E. Food Res. Int. 2006, 39, 383-393.
b) Kris-Etherton P.M., Harris W.S., Appel L.J. Circulation 2002, 106, 2747-2757.
c) Ramakrishnan V.V., Ghaly A.E., Brooks M.S., Budge S.M. Enz. Eng. 2013, 2, 115.
Poster 15
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
SEARCH FOR MOLECULAR MARKERS OF THERAPEUTIC
RESPONSE IN MALIGNANT MELANOMA THROUGH NGS: A
PROPOSAL OF ANTI-BRAF COMBINATION THERAPIES
Pisano M., Manca A., Palmieri G., Casula M., Cantara A., Dettori M. A.,
Fabbri D., Rozzo C.
Institute of Biomolecular Chemistry (ICB), National Research Council (CNR),
Traversa la Crucca, 3, 07100 Sassari, Italy
email: [email protected]; [email protected]
Introduction: Recent development of new targeted agents is giving viable
treatment options to melanoma therapies. Vemurafenib® specifically targets
BRAF-V600E mutation, a highly common alteration found in malignant melanomas
(MM). This targeted therapy is very effective on BRAF mutated MM patients but,
most of them quickly develops an acquired drug resistance, limiting the therapeutic
efficacy. We previously characterized the potential antitumor activity of several
compounds on MM cells using a series of cell lines derived from MM patients
showing different histopathological features (primary and/or metastatic, different
tumor stage and site of onset). Using NGS approaches, we started the molecular
characterization of Vemurafenib-resistant BRAF-mutated MM cell lines in
comparison with their drug-sensitive counterparts, in order to identify genetic
variants associated to the resistance. The evaluation of the antiproliferative activity
of some new compounds - like the curcumin-related D6 molecule - on
Vemurafenib-resistant MM cell lines is in progress.
Materials and Methods: The NGS testing was performed on cell lines using the
Ion-Torrent® technology. The validation of new markers will be carried out in vitro
(same cell lines) and in vivo (MM paraffinated tissue samples) by Sanger
sequencing. BRAF-mutated MM cell lines were treated with Vemurafenib (20 M
to 5 M) to isolate resistant clones. These were treated with a combination of
Vemurafenib and the curcumin-related D6 compound, before performing MTT
proliferation assays.
Results and discussion: A genetic heterogeneity was observed after
characterization of several primitive and metastatic MM cell lines by cancer gene
mutation profiling. Among them, four paired BRAF-mutated primitive and
metastatic MM cell lines were selected to isolate Vemurafenib-resistant clones.
These showed to be sensitive to D6 treatments, thus suggesting a combination
therapy using Vemurafenib and D6. NGS results are being evaluated in order to
identify alterations acting as markers for predicting the antiproliferative response
following this treatment. Moreover, they will allow a better understanding of the
molecular mechanisms underlying the development of resistance to Vemurafenib
in MM.
Conclusion: Our preliminary results suggest the feasibility of Vemurafenib-D6
combination treatment in drug-resistant MM. Our aim is to identify and validate
new molecular markers associated to therapeutic response in order to obtain a
better classification of the cases to be addressed to the treatment.
Poster 16
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
LIPASE CATALYSED RESOLUTION OF
PROPAGYL ACRYLATES, AS SYNTHONS IN ENYNE METATHESIS
Lambusta D.1, Gambera G.1, D’Annibale A.2, Nicolosi G.1
1
Institute CNR of Biomolecular Chemistry, via Paolo Gaifami 18, 95126 Catania
email: [email protected]
2
Chemistry Department, University “La Sapienza”, P.le Aldo Moro 5, 00185 Roma
Five and six-membered lactones are widely present as structural subunits in a
large number of natural compounds, showing important biological activities. 1
Since the ring-closing enyne metathesis (RCEYM) reaction represents a powerful
way to obtain cyclic structures, we were interested in exploring its potential in the
synthesis of naturally occurring fragrances detectable in spirits featuring a chiral
lactone ring system. Exploiting the enantiostereopreference of lipases when used
in organic solvents, we developed an enzymatic resolution of propargyl acrylates
racemic mixtures, in order to obtain enantiomerically pure esters subsequently
undergoing a RCEYM reaction, in order to prepare enantiopure chiral lactones.
In a first stage, we concentrated our attention on four propargyl alcohols (1a-d).
The enzymatically catalyzed selective acetylation of each corresponding alcohol
racemate was attempted, but unfortunately the use of lipases from Candida
antarctica, Mucor miehei , Porcine Pancreas, and Pseudomonas fluorescens, was
unsuccessful. We then moved our attention to the enzymatic alcoholysis of the
corresponding racemic acrylates (2a-d) with n-butanol using lipase from C.
Antarctica immobilized on support (Novozyme® 435) as the biocatalyst, in tert-butyl
methyl ether.
In these conditions the alcoholysis occured for all the four esters with high
enantioselectivity (E>200), and residual acrylates 2a-d were recovered with high
enantiomeric excess. Both the alkyl-substituted derivative 2b and aryl derivative
2d, showed to react faster, leading in 24h to the total conversion of the recognised
enantioform. According to our expectations, Lipase from C. antarctica brought to a
marked R stereopreference.
1
References: Kanchiswamy C.N.; Malnoy M.; Maffei M. E; Front. Plant Sci. 2015,6, 1-23
Poster 17
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
SYNTHESIS AND STUDY OF NEW HYDROXYLATED BIPHENOL
DERIVATIVES AS POTENTIAL TYROSINASE INHIBITORS
Dettori M.A.1, Fabbri D.1, Fois X.2,
Rocchitta G.3, Dallocchio R.1, Dessì A.1, Serra P.A.3,
Pantaleoni R.2,4 and Delogu G.1
1
CNR- Istituto di Chimica Biomolecolare, 2 CNR- Istituto per lo Studio degli
Ecosistemi,Traversa La Crucca 3- 07100 Sassari, Italy .3 UNISS- Dipartimento
di Medicina Clinica e Sperimentale, V.le S. Pietro 43/b- 07100 Sassari, Italy.
4
UNISS- Dipartimento di Agraria- Sezione di Entomologia,Via Enrico De Nicola
9- 07100 Sassari, Italy.
email: [email protected]
The role of melanin is to protect the skin from ultraviolet (UV) damage by
absorbing UV sunlight Melanin is one of the most widely distributed pigments
and is found in bacteria, fungi, plants and insects. Tyrosinase (polyphenol
oxidase, E.C. 1.14.18.1) and laccase (phenol oxidase, E.C. 1.10.3.2) are, the
keys enzymes in melanin biosynthesis, These inhibitors are also known to be
useful in cosmetics as whitening agents.a The involvement of laccase in cuticle
sclerotization or tanning is essential to insect survival.b
Hydroxylated biphenyls with a C2-symmetry axis have attracted considerable
interest because of the biological activity exerted by a number of natural
occurring compounds containing this moiety.d It is known that 4,4’dihydroxybiphenyl 1 has been shown to be a potent inhibitor of tyrosinase.c
Our study is aimed to prepare new biphenol derivatives as potential inhibitors of
melanin production starting from compound 1. Herein are reported some of the
new synthesized biphenyls in which, with the aim to improve water-solubility,
one hydroxyl is protected by aminoacid (2) gallic acid (3) and sugar (4 and 5)
unit, respectively. Compounds 6-8 belong to the class of C-prenylated
biphenyls.
Biological evaluation on PC12 cells, phytoiatric tests and docking studies of the
new derivatives were carried on.
Acknowledgments: This work has been supported by Sardinia Autonomous Region - L.R. 7
Augus2007, n. 7.
References: a) Chang. T. S. Int. J. Mol. Sci. 2009, 10, 2440-2475. b) Dittmer, N. T.; Kanost, M.
R. Insect Biochem. Mol. Biol. 2010, 40, 179-188. c) Kim Y. J.; No J. K.; Lee J. K. and Chung H.
Y. Bio. Pharm. Bull. 2005, 28, 323-327. d) Nicolaou, K. C.; Christopher .C. N.; Brȁse B. S.;
Nicholas N. Angew. Chem. Int. Ed., 1999, 38, 2096-2152.
Poster 18
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
PHYTOCHEMICAL STUDIES OF SOME
MARINE AND TERRESTRIAL MEDITERRANEAN PLANTS
Ciavatta M. L.1, Bitam F.2, Smadi A.3, Carbone M.1, Boumaraf M.4, Villani G.1,
Gavagnin M.1
1
Istituto di Chimica Biomolecolare-CNR, Via Campi Flegrei 34, I-80078 Pozzuoli (Naples)
2
Département de Pharmacie, Faculté de Médecine, Université de Batna (05000), Algérie
3
Laboratoire de Chimie et Chimie de l’Environnement (L.C.C.E), Département de Chimie, Faculté
des Sciences, Université de Batna (05000), Algérie
4
Department of Chemistry, Faculty of Exact Sciences, University of Mentouri-Constantine, 25000
Constantine, Algeria
email: [email protected]
Phytochemicals are biologically active, naturally occurring chemical compounds found in
plants, which are well-known to provide health benefits for humans.a In general, these
chemicals are produced to protect plant cells from environmental hazards such as
pollution, stress, drought, UV exposure and pathogenic attack. b They protect plants from
disease and damage and contribute to the plant’s colour, aroma and flavour.
Phytochemicals are usually accumulated in different parts of the plants, such as roots,
stems and leaves and belong to different structural types.c
Angiosperms are the largest group of plants on Earth (80% of the all known living plants)
and include aquatic and land species. They also are a rich source of new molecules with
interesting pharmacological properties that could be used as lead compounds for the
development of new drugs.d
In the frame of a scientific collaboration with researchers from Algerian universities we
have recently started a phytochemical screening on plants belonging to the Mediterranean
flora. Three angiosperms have been investigated: Cymodocea nodosa, a seagrass
collected in the Gulf of Naples, and Eryngium triquetrum and Pulicaria undulata, both
plants picked up from Algerian desert region.
A series of metabolites belonging to distinct structural classes have been isolated and
characterised by NMR and mass techniques. Phenolic and terpenoidic compounds have
been identified from C. nodosa, whereas polyacetylenes have been isolated for the first
time from E. triquetrum. Finally, novel terpenes belonging to the humulene class, have
been characterised from the lipophilic extract of P.undulata.
References:
a)
b)
c)
d)
Hasler C.M.; Blumberg J.B. J. Nutr.1999, 129: 756S-757S.
Piasecka A.; Jedrzejczak-Rey N.; Bednarel P. NewPhytologist2015, 206, 948-964..
Maag D.; ErbM.; Kollner T. G.; Gershenzon J. Bioessey2014, 37, 167-174.
Harvey A. L.; Edrada-Ebel R.; Quinn R. J. Nature Review Drug Discovery 2015, 14, 111-129.
Poster 19
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
DETERMINATION OF AVERMECTINES IN RAW WOOL BY LIQUID
CROMATOGRAPHY COUPLED TO ORBITRAP MASS
SPECTROMETRY
Azara E.1, Bortolu S.2, Lo Curto P.3, Duce P.2
1
Institute of Biomolecular Chemistry, National Research Council, Sassari, Italy
Institute of Biometeorology, National Research Council, Sassari, Italy
3
Clinical Epidemiology and Medical Statistics Unit, University of Sassari, Italy
email: [email protected]; [email protected]
2
The wool is a biodegradable renewable resource and, due to its complex chemical
composition and physical structure, can find various value-added applications (e.g.
thermal and acoustic insulation material, agricultural amendment, biomedical
polymers, etc.). In order to investigate if wool fibre would be a significant route of
human and environmental exposure, we developed a method to assess the
presence of drugs residues, deriving from veterinary practices.
Avermectins are macrocyclic lactones, which exhibit high anthelmintic, insecticide
and acaricide activity. This class of compounds includes ivermectine (IVE),
abamectine (ABA), emamectine (EMA), doramectine (DORA), eprinomectine
(EPRI), moxidectina (MOXI). These molecules and their transformation products
persist in the environment and can be toxic to no target organisms.
The aim of this work was to identify and quantify Avermectins in Sarda sheep wool
by High Liquid Chromatography coupled to Orbitrap mass spectrometry (LC-ESIMS/MS). Microwave assisted extraction (MAE), an environmentally friendly
method, was optimized with regard to the solvent type amount of solvent and
duration of extraction time. Since wool is a complex matrix with high protein and
fat content, which often interfere in analytical procedure, SPE Oasis hydrophilic
lipophilic balance (HLB) cartridges and QuEChERS extraction method were
compared as cleanup step. Chromatographic separation was achieved on a
Kinetex C18 column using a linear gradient of methanol-Ammonium Formiate 1
mmol containing 0.1% formic acid. The Q Exactive was equipped with heated
electrospray ionization source and operated in positive ionization mode. Orbitrap
spectrometer operated in targeted-MS/MS mode with precursor ion fragmentation
in HCD cell. The Full MS acquisition was performed with resolution power 70000
FWHM for parent ions and 17500 for the fragment ions with mass accuracy of 5
ppm. The calibration curves were linear with R2 always exceeding 0.99. The limit
of detection ranged from 0.5 to 2 ng/g.
Acknowledgment: Research funded by Med-L@ine project, granted by the Cross Border
Cooperation Programme Italy-Maritime 2013-2015
References: a) Rubies, A.; Antkowiiak, M.; Granados M.; Companyo R.; Centrich F. Food Chem
2015, 181, 57-63 b) Raich-Montiu, J.; Prat, M.D.; Granados M. Analytica Chimica Acta 2011, 697,
32-37
Poster 20
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
Presenting authors
Alberico E.
Andreotti G.
Azara E.
Biondi B.
Burreddu P.
Ciavatta M. L.
Cosseddu A.
Culeddu N.
Cutignano A.
Dallocchio R.
Dessì A.
Dettori M.A.
Drago C.
Fabbri D.
Fenude E.
Gavagnin M.
Granata G.
Lambusta D.
Mecca T.
Morrone R.
Peluso P.
Pisano M.
Piscitelli F.
Rozzo C.
Ruzza P.
Sanfilippo C.
Sanna D.
Sechi B.
Solinas M.
Spanu P.
Sperlinga E.
Villano R.
Vitale R.M.
Zambrano V.
ICB – SS
ICB – NA
ICB – SS
ICB – PD
ICB – SS
ICB – NA
ICB – SS
ICB – SS
ICB – NA
ICB – SS
ICB – SS
ICB – SS
ICB – CT
ICB – SS
ICB – SS
ICB – NA
ICB – CT
ICB – CT
ICB – CT
ICB – CT
ICB – SS
ICB – SS
ICB – NA
ICB – SS
ICB – PD
ICB – CT
ICB – SS
ICB – SS
ICB – SS
ICB – SS
ICB – CT
ICB – SS
ICB – NA
ICB – SS
Poster 5
Poster 12
Poster 20
Oral 6
Oral 4
Poster 19
Poster 6
Oral 10
Oral 9
Poster 1
Poster 1
Poster 18
Oral 11
Oral 2
Poster 2
Oral 12
Oral 8
Poster 17
Poster 11
Poster 15
Poster 3
Poster 16
Oral 3
Poster 16
Poster 9
Poster 13
Poster 14
Poster 4
Oral 5
Oral 1
Poster 8
Oral 7
Poster 10
Poster 7
2° Convegno dell'Istituto di Chimica Biomolecolare del CNR
Author Index
Alberico E. (Poster 5)
Amodeo P. (Poster 10)
Andreotti G. (Poster 12)
Azara E. (Poster 20)
Baglieri A. (Poster 15)
Battistini L. (Oral 4, Poster 7)
Baumann W. (Poster 5)
Beller M. (Poster 5)
Biondi B. (Oral 6, Poster 9)
Bisogno T. (Oral 3)
Bitam F. (Poster 19)
Bortolu S. (Poster 20)
Bortolus M. (Oral 6)
Boumaraf M. (Poster 19)
Brindani N. (Poster 7)
Bubacco L. (Poster 9)
Burreddu P. (Oral 4)
Cabrero-Antonino J. R. (Poster 5)
Cadoni R. (Poster 1)
Calderan A. (Poster 9)
Cantara A. (Poster 16)
Carbone M. (Oral 12)
Carbone M. (Poster 10, Poster 19)
Carcelli M. (Poster 1)
Casiraghi G. (Oral 4, Poster 7)
Casula M. (Poster 16)
Chandgude A. L. (Oral 1)
Chessa M. (Oral 10)
Ciavatta M.L. (Oral 12, Poster 10, Poster 19)
Cimmaruta C. (Poster 12)
Citro V. (Poster 12)
Consoli G.M.L. (Oral 8)
Cosseddu A. (Poster 6)
Cubellis M.V. (Poster 12)
Culeddu N. (Oral 10)
Cunsolo F. (Poster 11)
Curti C. (Poster 7)
Cutignano A. (Oral 9)
D’Annibale A. (Poster 17)
D’Antona N. (Poster 15)
Dallocchio R. (Poster 1, Poster 18)
Dalzini A. (Oral 6)
De Zotti M. (Oral 6)
Deiana P. (Oral 10)
Deligia F. (Oral 1)
Dell’Amico L. (Poster 7)
Delogu G. (Oral 2, Poster 18)
Desiderio D. (Poster 10)
Dessì A. (Poster 1, Poster 18)
Dettori M.A. (Oral 2, Poster 16, Poster 18)
Dettori S. (Oral 10)
Di Marzo V. (Oral 3)
Doemling A. (Oral 1)
Drago C. (Oral 11)
Drexler H.-J. (Poster 5)
Duce P. (Poster 20)
Fabbri D. (Oral 2, Poster 16, Poster 18)
Fadda A. (Poster 14)
Felicità V. (Poster 10)
Fenude E. (Poster 2)
Florio T. (Poster 10)
Fois X. (Poster 18)
Formaggio F. (Oral 6)
Fuggetta M.P. (Oral 1)
Gambera G. (Poster 15, Poster 17)
Gavagnin M. (Oral 12, Poster 10, Poster 19)
Geraci C. (Oral 8)
Giordano C. (Oral 3)
Giunta D. (Poster 4)
Granata G. (Oral 8)
Greco D. (Poster 7)
Guo Y.-W. (Oral 12)
Hussain R. (Poster 9)
Junge H. (Poster 5)
Junge K. (Poster 5)
Kiss R. (Oral 12)
Lambusta D. (Poster 17)
Lo Curto P. (Poster 20)
Lo Nigro R. (Oral 8)
Luongo L. (Oral 3)
Maione S. (Oral 3)
Malandrino G. (Oral 8)
Manca A. (Poster 16)
Marchetti M. (Poster 4)
Masullo M. (Poster 10)
Mathieu V. (Oral 12)
Mecca T.(Poster 11)
Molinu M.G. (Oral 10)
Mollo E. (Oral 12, Poster 10)
Morra A. (Oral 1)
Morrone R. (Poster 15)
Naesens L. (Poster 1)
Napoli E. (Poster 8)
Nicolosi G. (Oral 11, Poster 15, Poster 17)
Pala N. (Poster 1)
Palmieri G. (Poster 16)
Pantaleoni R. (Poster 18)
Patti A. (Poster 13)
Peggion C. (Oral 6)
Pelosi G. (Poster 7)
Peluso P. (Poster 3)
Pinna L. (Poster 7)
Pisano M. (Poster 16)
Piscitelli F. (Oral 3)
Rassu G. (Oral 4, Poster 7)
Rispoli V. (Poster 10)
Rocchitta G. (Poster 18)
Rozzo C. (Poster 16)
2°Convegno dell'Istituto di Chimica Biomolecolare del CNR
Ruberto G. (Poster 8)
Ruzza P. (Poster 9)
Sanfilippo C. (Poster 13)
Sanna D. (Poster 14)
Santona M. (Oral 10)
Sartori A. (Oral 4, Poster 7)
Sechi B. (Poster 4)
Sechi M. (Poster 1)
Serra M. (Poster 14)
Serra P.A. (Poster 18)
Sgammato R. (Poster 10)
Siligardi G. (Poster 9)
Siracusa L. (Poster 8)
Smadi A. (Poster 19)
Solinas M. (Oral 5, Poster 4)
Spanu P. (Oral 1)
Sperlinga E. (Poster 8)
Stevaert A. (Poster 1)
Tessari I. (Poster 9)
Thellung S. (Poster 10)
Ulgheri F. (Oral 1)
Villani G. (Poster 19)
Villano R. (Oral 7)
Vitale R.M. (Poster 10)
Vite V. (Poster 2)
Zambrano V. (Poster 7)
Zanardi F. (Oral 4, Poster 7)