3 CONVEGNO ANNUALE ICB - CNR - Catania via Paolo Gaifami, 9 - … · 3 CONVEGNO ANNUALE ICB - CNR -...

3 CONVEGNO ANNUALE ICB - CNR - CATANIA VIA PAOLO GAIFAMI, 9 - 95126 28-30 settembre 2016

www.ct.icb.cnr.it Book of Abstracts | 3 CONVEGNO ANNUALE ICB 2



Come gather 'round people

Wherever you roam

And admit that the waters

Around you have grown

And accept it that soon

You'll be drenched to the bone

If your time to you

Is worth savin'

Then you better start swimmin'

Or you'll sink like a stone

For the times they are a-changin'.

€



MERCOLEDÌ 28 SETTEMBRE

12:00 - 12:45 Registrazione, Messaggi di benvenuto del Direttore e del Responsabile UOS di Catania

13:00 - 14:15 Pranzo

PRIMA SESSIONE

Tematica: SOSTANZE NATURALI IN BIOMEDICINA Moderatore: Laura Siracusa

PL1 14:30 - 15:15

Antonina Saija - Dipartimento di Scienze Chimiche, Biologiche, Farmaceutiche ed Ambientali, Università di Messina. “Polifenoli di origine vegetale e malattie non trasmissibili: da antiossidanti a modulatori del network di segnale cellulare, benefici, problemi e prospettive”

OC1 15:15 - 15:35

Di Mauro M.D., Tomasello B., Morrone R., Biondi D., Renis M., D'Antona N. “Recovery of high-added-value compounds from olive mill wastewater for cosmetic and ophthalmic applications”

OC2 15:35 - 16:05

Petrosino S., Manzo E., Allarà M., Tinto F., Pagano D., Verde R., Schiano Moriello A., De Petrocellis L., Di Marzo V. “An innovative potential bioregulator developed from palmitoylethanolamide”

16:05 - 16:45 Pausa caffè

OC3 16:45 - 17:05

Iannotti F.A., Pagano E., Adinolfi S., Guardiola O., Gazzerro E., Capasso R., Minchiotti G., Di Marzo V. “Study of the expression profile and pharmacological role of the CB1 receptor in Duchenne Muscular Dystrophy (DMD) muscles: a new opportunity to reinforce muscle repair and locomotor activity”

17:05 - 18:30 Riunione plenaria - Comunicazioni del Direttore - Discussione

18:30 - 19:30 Riunione del CdI Serata libera



GIOVEDÌ 29 SETTEMBRE

SECONDA SESSIONE

Tematica: BIOPROCESSI PER LA SOSTENIBILITA’ E LA PRODUZIONE ENERGETICA

Moderatore: Giuseppina Andreotti

PL2 9:30 - 10:15

Salvatore L. Cosentino - Dipartimento di Agricoltura, Alimentazione e Ambiente - Di3A, Università di Catania. ”Il contributo delle biomasse vegetali alla produzione di energia e alla sostenibilità ambientale”

OC4 10:15 - 10:35

d’Ippolito G., Fontana A. “Bioprocessi sostenibili da microrganismi marini per la produzione di bioenergia e prodotti funzionali: dalla natura alle applicazioni industriali”

OC5 10:35 - 10:55

Morrone R., D’Antona N. “Biocatalyzed kinetic resolution of racemic hexaprofen by efficient irreversible esterification”


11:30 - 13:00 In simultanea riunioni SICI: CARFIL, PREDIR, CAPRI, RIND e 1a Sessione Poster

13:00 - 14:00 Pranzo

TERZA SESSIONE

Tematica: NUOVE FRONTIERE NELLA SINTESI ORGANICA Moderatore: Giovanna Delogu

PL3 14:15 - 15:00

Giovanni Maria Pitari - Department of Pharmacology and Experimental Therapeutics, Thomas Jefferson University, Philadelphia, Pennsylvania – Vera Salus Ricerca S.r.l., Viagrande (CT). “Dalla Teoria alla Pratica: Translational Research & Beyond”

OC6 15:00 - 15:20

Foti M.C., Slavova-Kazakova A., Rocco C., Kancheva V.D. “Kinetics of curcumin oxidation with DPPH• and lipid peroxyl radicals”

OC7 15:20 - 15:40

Patti A., Pedotti S. “Ferrocene derivatives: synthesis and applications”

OC8 15:40 - 16:00

Gambera G., Greco G., D’Antona N., Nicolosi G., Corda D. “Synthesis of analogues and derivatives of glycerophosphoinositol (GroPIns)”

16:00 - 16:30 2a Sessione Poster

19:00 - 20:30 Visita del Monastero dei Benedettini

21:00 Cena sociale



VENERDÌ 30 SETTEMBRE

QUARTA SESSIONE

Tematica NANOTECNOLOGIE

Moderatore: Andrea Calderan

PL3 9:30 - 10:15

Assunta Borzacchiello - Istituto del CNR per i Polimeri, Compositi e Biomateriali, Napoli. “Smart hydrogels and nanoparticulate systems for drug delivery and regenerative medicine applications”

OC9 10:15 - 10:35

Mecca T., Cunsolo F. “Calix[8]arene polyelectrolytes: a useful class of pH-responsive hydrogelators with new interesting features”

OC10 10:35 - 10:55

Di Bari I., Consoli G.M.L., Sortino S. “Calix[4]arene-based photoresponsive nanosystems for antibacterial applications”


11:15 - 11:35

Giuseppe Manuele - Confindustria Catania, Presidente della Sezione Chimici e Chimico Farmaceutici, e A.D. di MAPLAD S.r.l., Catania. “La ricerca applicata alla base di ogni innovazione industriale”

11:35 - 11:55

Salvo Fallica - Business Incubation Manager – Vulcanìc, Catania. “Startup, Spin-off e PMI innovative: Il ruolo degli Incubatori di Impresa a supporto del nuovo tessuto imprenditoriale europeo”

11:55 - 12:15 Comunicazione del vincitore del bando interno ICB ed esposizione del Progetto

12:15 - 15:30 Conclusioni, chiusura lavori Pranzo di commiato



PROF.SSA ANTONINA SAIJA Dipartimento di Scienze Chimiche, Biologiche, Farmaceutiche ed Ambientali, Università di

Messina.

“POLIFENOLI DI ORIGINE VEGETALE E MALATTIE NON TRASMISSIBILI: DA ANTIOSSIDANTI A MODULATORI DEL NETWORK DI SEGNALE CELLULARE, BENEFICI, PROBLEMI E PROSPETTIVE”

PROF. SALVATORE L. COSENTINO Dipartimento di Agricoltura, Alimentazione e Ambiente - Di3A, Università di Catania.

“IL CONTRIBUTO DELLE BIOMASSE VEGETALI ALLA PRODUZIONE DI

ENERGIA E ALLA SOSTENIBILITÀ AMBIENTALE”

DOTT. GIOVANNI MARIO PITARI Department of Pharmacology and Experimental Therapeutics, Thomas Jefferson University, Philadelphia, Pennsylvania – Vera Salus Ricerca S.r.l., Viagrande (CT).

“DALLA TEORIA ALLA PRATICA: TRANSLATIONAL RESEARCH &

BEYOND”



DOTT.SSA ASSUNTA BORZACCHIELLO Istituto del CNR per i Polimeri, Compositi e Biomateriali, Napoli.

“SMART HYDROGELS AND NANOPARTICULATE SYSTEMS FOR DRUG DELIVERY AND REGENERATIVE MEDICINE APPLICATIONS”

ING. GIUSEPPE MANUELE Confindustria Catania, Presidente della Sezione Chimici e Chimico Farmaceutici, e A.D. di MAPLAD S.r.l., Catania.

“LA RICERCA APPLICATA ALLA BASE DI OGNI INNOVAZIONE

INDUSTRIALE”

DOTT. SALVO FALLICA Business Incubation Manager – Vulcanìc, Catania.

“STARTUP, SPIN-OFF E PMI INNOVATIVE: IL RUOLO DEGLI

INCUBATORI DI IMPRESA A SUPPORTO DEL NUOVO TESSUTO

IMPRENDITORIALE EUROPEO”



Di Mauro M.D.a,b, Tomasello B.b, Morrone R.a, Biondi D.a, Renis M.b, D'Antona N.a

a CNR-Institute of Biomolecular Chemistry, via Gaifami 18, 95126, Catania

b Department of Drug Sciences, Biochemistry Section, University of Catania, viale Andrea Doria 6, 95125, Catania

[email protected]

The olive oil industry produces huge amounts of olive mill wastewater (OMWW) as the main byproduct of the olive oil extraction process [1]. Since a long time, OMWW have represented an economic and environmental problem due their antimicrobial, ecotoxic and phytotoxic properties, related to the high chemical content [2]. Nowadays OMWW are considered a potential resource of valuable compounds, such as sugars, minerals and polyphenols, that can find interesting applications in several industrial fields. Indeed, sugars and minerals are hygroscopic components of the natural moisturizing factor, playing a crucial role in maintaining the skin hydration, that find a great deal of attention in the cosmetic formulation strategies [3]. On the other hand, polyphenols are well known for their antioxidant and anti-inflammatory activities; in particular, hydroxytyrosol, the most abundant polyphenol in OMWW, is able to scavenge free radicals, to protect against oxidative damage, to inhibit the proinflammatory enzymes and to modulate the release of proinflammatory cytokines [4-5]. Taking into account these properties, ophthalmic formulations containing polyphenolic compounds may represent an important therapeutic solution for all the pathologies of the ocular surface with an inflammatory component. In this talk, we will report an innovative green methodology to recover chemically distinct pools of compounds from OMWW through an adsorption/desorption process, and their application to the development of cosmetic and nutraceutical ophthalmic formulations. Moreover, the strategy was scaled up and a pre-industrial pilot plant prototype was developed too. Acknowledgment: Research was funded by the Sicily Region in the framework of the European Regional Development Fund (Progetto N.1422 PO FESR Sicilia 2007/2013 Linea di intervento 4.1.1.1). References:

1. Deeb, A.A., Fayyad, M.K., Alawi, M.A. Chromatogr. Res. Int. 2012, 1-8. 2. Magdich, S., Jarboui, R., Rouina, B.B., Boukhris, M., Ammar, E. Sci. Total Environ. 2012, 430, 209-216. 3. Verdier-Sévrain, S., Bonté, F. J. Cosmet. Dermatol. 2007, 6, 75-82. 4. Hu, T., He, X., Jiang, J., Xu, X. J. Agric. Food Chem, 2014, 62, 1449-1455. 5. Grasso, S., Siracusa, L., Spatafora, C., Renis, M., Tringali, C. Bioorg. Chem. 2007, 35 (2), 137-152.

RECOVERY OF HIGH-ADDED-VALUE COMPOUNDS FROM OLIVE MILL WASTEWATER FOR COSMETIC AND OPHTHALMIC APPLICATIONS

mailto:[email protected]



Petrosino S.1,2,3, Manzo E.1, Allarà M.1,2,3, Tinto F.1, Pagano D.1, Verde R.1,2,

Schiano Moriello A.1,2,3, De Petrocellis L.1,2, Di Marzo V.1,2

[email protected]

1Institute of Biomolecular Chemistry, CNR, via Campi Flegrei 34, 80078, Pozzuoli (NA), Italy; 2Endocannabinoid Research Group; 3Epitech Group S.p.A., via Einaudi 13, 35030,

Saccolongo (PD), Italy.

Introduction: Palmitoylethanolamide (PEA) is a lipid mediator used in the clinic for its anti-inflammatory and analgesic propertiesa,b,c. In order to improve its bioavailability we designed, realized and developed an adduct between PEA (the active molecule) and a carrier, glucuronic acid. The bioregulator, Glucuropea, was characterized in vitro and its anti-inflammatory activity studied in an experimental model of inflammation in vitro and in a model of colitis in mice. Methods: Human keratinocyte (HaCaT) cells were treated with Glucuropea, and the levels of PEA and endocannabinoids (anandamide, 2-arachidonoylglycerol) measured. The effect of Glucuropea, per se or in cells concomitantly treated with 2-AG, at human transient receptor potential vanilloid type-1 (TRPV1) was also assessed by measuring intracellular Ca2+ in human embryonic kidney (HEK-293) cells over-expressing TRPV1. The anti-inflammatory activity of Glucuropea in vitro was investigated in HaCaT cells by assessing its effect on polyinosinic-polycytidylic acid-induced release of the pro-inflammatory chemokine, monocyte chemotactic protein-2 (MCP-2). The anti-inflammatory activity in vivo was evaluated in dinitrobenzene sulfonic acid (DNBS)-treated mice, which replicate some of the features of colitis. Results: Treatment with Glucuropea, as compared to the same concentration of PEA, resulted in significantly higher amounts not only of PEA but also of 2-AG in HaCaT cells. Glucuropea only slightly enhanced 2-AG activation of TRPV1, but significantly increased 2-AG-induced TRPV1 desensitization to capsaicin. Glucuropea inhibited the release of the pro-inflammatory chemokine MCP-2 from stimulated HaCaT cells. Glucuropea was as efficacious as PEA at reducing the signs of colitis in DNBS-treated mice when using the same dose (which, however, corresponds to a lower molar amount). Conclusions: Glucuropea is a novel anti-inflammatory bioregulator, able to enhance both PEA levels and actions on endocannabinoid signaling in cellular systems.

Acknowledgement: Research funded by Progetto Operativo Nazionale (PON01_02512). References:

1. Petrosino, S., Iuvone, T., Di Marzo, V. N-palmitoyl-ethanolamine: Biochemistry and new therapeutic opportunities. Biochimie. 2010, 92, 724-727.

2. Petrosino, S., Cristino, L., Karsak, M., Gaffal, E., Ueda, N., Tüting, T., Bisogno, T., De Filippis, D., D'Amico, A., Saturnino, C., Orlando, P., Zimmer, A., Iuvone, T., Di Marzo, V. Protective role of palmitoylethanolamide in contact allergic dermatitis. Allergy. 2010, 65, 698-711.

3. Petrosino, S., Schiano Moriello, A., Cerrato, S., Fusco, M., Puigdemont, A., De Petrocellis, L., Di Marzo, V. The anti-inflammatory mediator palmitoylethanolamide enhances the levels of 2-arachidonoyl-glycerol and potentiates its actions at TRPV1 cation channels. Br. J. Pharmacol. 2016, 173, 1154-1162.

AN INNOVATIVE POTENTIAL BIOREGULATOR DEVELOPED FROM PALMITOYLETHANOLAMIDE




Iannotti F.A.1, Pagano E.2, Adinolfi S.1, Guardiola O.3, Gazzerro E.4, Capasso R.2, Minchiotti G.3 and Di Marzo V.1

[email protected]

1 Endocannabinoid Research Group, Institute of Biomolecular Chemistry (ICB), National Council of Research (CNR) , Pozzuoli (NA) Italy; 2 Department of Pharmacy, University of Naples Federico II; 3Gaslini Children's Hospital, L.go Gaslini 5 16147 Genova; 4Institute of Genetics and Biophysics 'A. Buzzati-Traverso', CNR, Naples, Italy. Duchenne’s muscle dystrophy (DMD) represents the most frequent form of hereditary myopathy. It is caused by mutations in the X-linked gene (locus Xp21.2) encoding for the structural protein dystrophin. In healthy skeletal and cardiac muscles, dystrophin plays a critical structural role by being part of the dystrophin-glycoprotein complex (DGC) that physically connects the cytoskeleton to the surrounding extracellular matrix through the cell membrane. Therefore, the loss of dystrophin function caused by large intragenic deletions (65% of the cases), or intragenic duplications (6–10% of the cases) or point mutations associated to other sequence variations (30–35% of the cases), leads to progressive and irreversible muscle wasting and weakness. Since the dystrophin gene is located on the X chromosome, the disease mostly affects young boys with a rate of approximately 1:3500 (1). Recently, we have shown that the muscle levels of 2-AG are decreased both during myotube formation in vitro from murine C2C12 myoblasts, and during mouse muscle development. Moreover, we reported that in both murine and primary human myoblasts, the antagonism of CB1 receptor by rimonabant (SR141716), rather than its agonism by ACEA, promotes myoblast differentiation into myotubes (2).

Therefore, the purpose of this study was to study the role of CB1 receptors in skeletal muscles affected by Duchenne’s muscular dystrophy (DMD).

We found that, in muscle samples from dystrophic mice (DMDmdx) and DMD humans, CB1 transcripts show the highest degree of expression near the onset of disease, and then decline overtime. Similar changes were observed for Pax7, a key transcriptional factor regulating muscle regeneration through satellite cell activation. Bioinformatic analysis revealed the presence of putative Pax7 responsive sequences in murine and human CB1 genes. Chromatin immunoprecipitation and luciferase assays showed that PAX7 binds to and upregulates the CB1 gene, with higher efficacy in DMDmdx than healthy mouse muscles. Antagonism of CB1 with rimonabant promoted human healthy satellite cell and DMD myoblast differentiation in vitro, and increased the number of regenerated myofibers and reduced the levels of fibrosis and inflammation markers in dystrophic mice. In conclusion, our study uncovers a potential Pax7-CB1-engendered vicious cycle exacerbating defective satellite cell differentiation, and points to pharmacological antagonism of CB1 receptors as co-adjuvant therapy for muscular dystrophies. References:

1) Goyenvalle A, et al. (2011) Therapeutic approaches to muscular dystrophy Hum Mol Genet. 20(R1):R69-78

2) Iannotti et al. (2014) The endocannabinoid 2-AG controls skeletal muscle cell differentiation via CB1 receptor-dependent inhibition of Kv7 channels. Proc Natl Acad Sci U S A. 2014 Jun 17;111(24):E2472-81.

STUDY OF THE EXPRESSION PROFILE AND PHARMACOLOGICAL ROLE OF THE CB1 RECEPTOR IN DUCHENNE MUSCULAR DYSTROPHY

(DMD) MUSCLES: A NEW OPPORTUNITY TO REINFORCE MUSCLE REPAIR AND LOCOMOTOR ACTIVITY




Giuliana d’Ippolito, Angelo Fontana

Istituto di Chimica Biomolecolare ICB-CNR, Via Campi Flegrei 34, 80078 Pozzuoli (Napoli), Italy

[email protected]

La produzione di biomasse da microalghe marine, così come la produzione di idrogeno da parte di batteri termofili, sono singolarmente processi molto produttivi e a basso impatto ambientale per la produzione di vettori energetici rinnovabili. Il pieno utilizzo di queste tecnologie a livello industriale è però ancora frenato dagli alti costi di produzione. Al fine di ottenere un abbattimento dei costi ed un aumento della redditività è importante lavorare sull’ottimizzazione dei singoli processi e su un’effettiva ed efficiente integrazione di diversi sistemi biologici per permettere il loro pieno sfruttamento in maniera economicamente sostenibile.

Attraverso casi specifici affrontati nei nostri laboratori, si ripercorreranno le varie fasi che vanno dall’isolamento, la selezione e la coltivazione del microrganismo di interesse, fino allo sviluppo di un bio-processo potenzialmente appetibile a livello industriale. Inoltre verrà evidenziato come lo studio su basi chimiche del metabolismo microbico risulti cruciale per migliorare le rese dei processi.

References: d'Ippolito, G.; Dipasquale, L.; Vella, F.M.; Romano, I.; Gambacorta, A.; Cutignano, A.; Fontana A. Int J of Hydrogen Energy, 2010, 35, 2290. d'Ippolito, G.; Dipasquale, L.; Fontana, A. ChemSusChem. 2014 7(9), 2678. Dipasquale, L.; d'Ippolito, G.; Fontana, A. Int J of Hydrogen Energy, 2014 39 (10), 4857. Dipasquale, L.; d’Ippolito, G.; Gallo, C.; Vella, F.M.; Gambacorta, A.; Picariello, G.; Fontana, A. Int J of Hydrogen Energy 2012, 37, 12250. Dipasquale, L.; Adessi, A.; d'Ippolito, G.; Rossi, F.; Fontana, A.; De Philippis, R.; Appl Microbiol Biotechnol. 2015, 99(2), 1001. d'Ippolito, G.; Sardo, A.; Paris, D.; Vella, F.M.; Adelfi, M.G.; Botte, P.; Gallo, C.; Fontana, A.; Biotechnol Biofuels. 2015, 22,8,28.

BIOPROCESSI SOSTENIBILI DA MICRORGANISMI MARINI PER LA PRODUZIONE DI BIOENERGIA E PRODOTTI FUNZIONALI: DALLA

NATURA ALLE APPLICAZIONI INDUSTRIALI




Morrone R., D’Antona N.

CNR – Institute of Biomolecular Chemistry, Via Paolo Gaifami, 18 - 95126 Catania, Italy

[email protected]

The most widely used class of NSAIDs in the current therapy of inflammatory conditions is that of ‘profens’, 2-arylpropionic acids derivatives, non-selective inhibitors of cyclooxygenases.1 The profens are chiral drugs and the anti-inflammatory activity is mainly due to S enantiomer. The use of active form is essential to minimize the dose and to reduce toxicity and variability in the response. Furthermore taking into account the recommendations of the Federal and Drug Administration and of the European Medicines Agency that consider the R-, S-enantiomers and racemate as three different drugs, the access to both the pure enantioforms of a chiral molecule is mandatory for the evaluation of the specific properties. Among the non-marketed profens, hexaprofen [2-(4-cyclohexylphenyl) propionic acid] is a very attracting compound also for its antitumor activity.2

Lipases, under natural aqueous conditions, catalyze the hydrolysis of ester bonds. When used in organic solvent their behaviour is reversed, and the esterification reaction between an acid and a suitable nucleophile is catalyzed. This features combined with the broad substrate recognition, robustness, high efficiency and selectivity have made lipases as the class of biocatalysts most used in organic chemistry. Their ability to recognize chiral elements has been exploited in many processes of resolution of racemic mixtures including the profens.4

In our laboratory, we have developed strategies that allow efficient resolution of chiral acids by biocatalyzed irreversible esterification.5 Applying this methodology, we report here the resolution of hexaprofen by irreversible esterification in organic solvent catalyzed by lipase and the determination of absolute configuration. References:

1. Agranat, I.; Caner, H.; Caldwell, J. Nat. Rev. Drug Discov. 2002, 1, 753–68. 2. Mamytbekova, A.; Čepelak, V.; Grimova, J.; Řežabek, K.; Kuchař, M. Neoplasma 1989, 36, 327-332. 3. Sikora, A.; Siódmiak, T.; Marszałł, M.P. Chirality 2014, 26, 663-669 and references cited therein. 4. Morrone, R.; D'Antona, N.; Lambusta, D.; Nicolosi, G. J. Mol. Cat. B: Enzym. 2010, 65, 49–51.

BIOCATALYZED KINETIC RESOLUTION OF RACEMIC HEXAPROFEN BY

EFFICIENT IRREVERSIBLE ESTERIFICATION




Foti M.C.,a, Slavova-Kazakova A.,b Rocco C.a and Kancheva V. D.b

aIstituto di Chimica Biomolecolare del CNR, Via P. Gaifami, 18 – 95126 Catania (Italy);

bInstitute of Organic Chemistry with Centre of Phytochemistry,

Bulgarian Academy of Sciences, “Acad. G. Bonchev” Str., Bl.9 - 1113 Sofia (Bulgaria)

[email protected]

The observed kinetics of the reactions between dpph• and curcumin (1) or curcumin derivatives (2 – 5) in absolute ethanol at 25 °C are complex. The decay of dpph• at 517 nm in the presence of curcumin (1), 4-methylcurcumin (3), 4,4-dimethylcurcumin (4) and curcumin 4’-methyl ether (5) follows bi-exponential kinetics with a fast initial process followed by a slower process. These unusual kinetics are compatible with a two-step process in which an intermediate accumulates in a reversible step followed by an irreversible process. As in other similar cases,1 we have

hypothesised that the intermediate is a -stacked complex where a fast electron transfer process occurs. By comparing the kinetics of curcumin 4’,4”-dimethyl ether (2) (no phenolic OH), (5) (one phenolic OH) and (1) (two phenolic OHs), we have also deduced that the electron transfer is simultaneous to a proton transfer process from the phenolic OHs to the solvent. Curcumin is also able to retard the spontaneous autoxidation of triacylglycerols of sunflower oil

at 80 °C with an efficacy that, at a concentration of 1 mM, is comparable to that of -tocopherol.

References:

1. Foti et al. Org. Lett. 2011, 13, 4826-4829.

KINETICS OF CURCUMIN OXIDATION WITH DPPH• AND LIPID PEROXYL RADICALS




Patti A., Pedotti S.

Istituto di Chimica Biomolecolare, Via Paolo Gaifami, 18 – 95126 Catania

[email protected]

Ferrocene derivatives have attracted growing interest for their applications in the design of new materials with peculiar optical and redox properties,1 as ligands for asymmetric catalysis2 and promising candidates in drug development.3 In the course of our research we have explored the chemistry of ferrocenes and developed different routes to chiral ferrocenes. Some of them have been investigated for their catalytic activity and, recently, we successfully designed the first ferrocene-proline conjugate for its application in asymmetric organocatalytic aldol reactions in water medium.4

In agreement with the emerging bio-organometallic approach, we have also synthesized the ferrocenyl analogues of some bioactive compounds and a positive effect related with the presence of the ferrocene moiety was evidenced. In this context, our recent work involved the synthesis of two ferrocenyl analogues of clotrimazole, in which the metallocene fragment replaces one of the phenyl ring in the triarylmethane scaffold of the drug. The synthesis of target organometallic compounds exploited the direct reaction with imidazole on a suitable intermediate in the key step, providing an

additional example in the short list of nucleophilic substitutions on fully -substituted ferrocenes. The obtained ferrocene derivatives were then evaluated for their cell growth inhibitory

activity on two different human cancer cell lines and, in comparison with the parent drug, about two-fold increase of cytotoxicity on HT29 colorectal cancer cells was displayed, whereas comparable activity was measured against MCF-7 breast cancer cells.5

References:

1. Xiang, J.; Sato, K.; Tokue,H.; Oyaizu, K.; Ho, C.-L.; Nishide, H.; Wong, W.-Y.; Wei M. Eur. J. Inorg. Chem. 2016, 1030-1035.

2. Arrayás, R. G.; Adrio, J.; Carretero, J. C. Angew. Chem. Int. Engl. Ed. 2006, 45, 7674-7715. 3. Braga, S. S.; Silva, A. M. S. Organometallics, 2013, 32, 5626-5639. 4. Patti, A.; Pedotti, S. Eur. J. Org. Chem. 2014, 624-630. 5. Pedotti, S.; Ussia, M.; Patti, A.; Musso, N.; Barresi, V.; Condorelli, D. F. submitted

FERROCENE DERIVATIVES: SYNTHESIS AND APPLICATIONS




Gambera G.a, Greco G. a, D’Antona N. a, Nicolosi G. a, Corda D.b

aCNR – Institute of Biomolecular Chemistry UOS Catania, Via P. Gaifami, 18 - 95126 Catania

b CNR - Institute of Protein Biochemistry, Via P. Castellino 111 – 80131 Napoli

[email protected]

The hydrolytic activity of phospholipases A (PLA2, LPLA1) on membrane phosphoinositides generates the formation of glycerophosphoinositols: glycerophosphoinositol (GroPIns), glycerophosphoinositol 4-phosphate (GroPins4P) and glycerophosphoinositol 4,5-biphosphate (GroPins4,5P2).

1

Glycerophosphoinositols are small hydrophilic molecules, hydrolyzed by the action of glycerophosphodiesterases (GDEs) into their inositol and glycerol components;2 they are involved in several regulatory pathways related to cell proliferation, motility and invasion.3

It has been demonstrated that GroPIns is able to stimulate osteoblasts and thyroid cell growth, and to inhibit the migration of human melanoma and human mammary carcinoma cells through the extracellular matrix (ECM):3 these findings suggest a role of GroPIns as a mediator for cells proliferation and a possible use in cancer treatment.

Unfortunately, to date, the molecular targets of these molecules are still uncertain.3

This work have been focused on the synthesis of molecular tools to investigate the mechanism of action of glycerophosphoinositols: analogues of GroPIns have been synthesized pursuing the stabilization towards the action of GDEs and to study the influence of those modification on its biological activity; moreover, with the aim of following its cellular distribution, fluorescent GroPins derivatives have been synthesized.4

Acknowledgment: Research funded by PON 01_00862 “Una Piattaforma Tecnologica Integrata per lo Sviluppo di Nuovi Farmaci per Malattie Rare” (Ministry of University and Research, Italy). References:

1. a) Corda, D., Iurisci, C. and Berrie, C.P. Biochim. Biophys. Acta 2002, 1582, 52–69; b) Mariggio`, S., Sebastia`, J., Filippi, B.M., Iurisci, C., Volonte´, C., Amadio, S., De Falco, V., Santoro, M. and Corda, D. FASEB J. 2006,. 20, 2567–2569.

2. Zheng, B., Chen, D., Farquhar, M.G.. Proc. Natl. Acad. Sci. U.S.A. 2000, 97, 3999–4004; b) Corda, D., Kudo, T., Zizza, P., Iurisci, C., Kawai, E., Kato, N., Yanaka, N., Mariggio` , S. J. Biol. Chem. 2009, 284, 24848–24856.

3. Corda, D., Zizza, P., Varone, A, Karol S. Bruzik, K-S, Mariggio`, S. Biochem. Soc. Trans. 2012, 40, 101–107 and references cited therein.

4. Greco, G., D’Antona, N., Gambera, G., Nicolosi, G. Synlett. 2014, 25, 2111-2114.

SYNTHESIS OF ANALOGUES AND DERIVATIVES OF GLYCEROPHOSPHOINOSITOL (GROPINS).




Mecca T., Cunsolo F. CNR-Istituto di Chimica Biomolecolare, UOS of Catania,

Via P. Gaifami, 18 - 95126, Catania [email protected]

Nowadays, supramolecular hydrogelators are considered of great interest in biomolecular field due to their various applications in cell culture and modulation, tissue engineering, imaging, and drug delivery.1 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.2 The supramolecular hydrogel structure is mainly mediated by electrostatic interactions between charged and uncharged polar groups on the calix[8]arene scaffolds. A correct balance between hydrophilicity and hydrophobicity of the molecules is also necessary for the gelification. In order to evaluate the utility of these hydrogels, basic pH-responsive calix[8]arene hydrogelators, has been studied for the loading and release capacity towards steroid and non-steroid anti-inflammatory drugs, while 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.3 Finally, a fully aromatic functionalization of the calix[8]arene scaffold afforded a compound whose pH of gelification can be tuned in a wide range by the simple addition of variable amount of a polyamine. This feature is, at the best of our knowledge, not yet reported in literature, and applications are still to be discovered and explored.

pH variations

Acknowlegment: Research funded by PON R&C 02_00355_2964193 (MIUR, Rome).

References:

1. Du, X.; Zhou, J.; Shi, J.; Xu, B. Chem. Rev. 2015,115,13165-13307. 2. Mecca, T.; Messina, G. M. L.; Marletta, G.; Cunsolo, F. Chem. Commun. 2013, 49, 2530-2532. 3. Naksuriya, O.; Okonogi, S.; Schiffelers, R. M.; Hennink, W. E. Biomaterials 2014, 35, 3365-3383.

CALIX[8]ARENE POLYELECTROLYTES: A USEFUL CLASS OF PH-RESPONSIVE HYDROGELATORS WITH NEW INTERESTING FEATURES.




Di Bari I.,a Consoli G.M.L.,b Sortino S.a

a Department of Drug Science, University of Catania, V.le A. Doria 6, 95125, Catania, Italy; b CNR-Institute of Biomolecular Chemistry, Via P. Gaifami, 18 – 95126 Catania, Italy.

[email protected]

In the search of innovative and more effective antimicrobial strategies to fight infections affected by antibiotic resistance phenomena, the use of photoresponsive agents able to trigger cytotoxic species, such as nitric oxide (NO) and singlet oxygen (1O2), eliciting oxidative damage, cell death, and biofilm dissolution is particularly attractive. Light-controlled generation, diffusion in the cellular environment over short distances (<200 μm), short half-life in blood, and multitarget action are features that confine the region of action of 1O2 and NO reducing undesired side effects and minimize drug resistance problems. Nevertheless, many photoresponsive agents have a hydrophobic nature, which favors their aggregation in aqueous medium and strongly precludes the photochemical behavior.

Nanotechnology is a valid support to overcome these drawbacks. The entrapment in nanostructured systems can enhance water solubility, protection from degradation, bioavailability, local concentration and site-specific delivery of photoresponsive agents.1 Calix[n]arene are interesting scaffolds in supramolecular chemistry, synthetic versatility, good biocompatibility, and capability to self-assembly in well-defined nanostructures have opened perspectives for applications in biomedical field.2

Here we report the first example of a self-assemblying amphiphilic calixarene decorated with homing choline groups, able to act as nanocontainer, nanoreactor, and potentially nanocarrier for NO donor and/or photosensitizers.

Figure 1. Schematic representation of the calixarene-based photoresponsive nanosystem.

The nanostructured systems generated NO and/or 1O2 upon to irradiation with visible light

and the light-stimulated bactericidal effect was successfully proved on Gram positive and Gram negative bacteria. References:

1. A. Fraix, N. Marino, S. Sortino, Topics Curr. Chem., 2016, 370, 225. 2. S. B. Nimse, T. Kim, Chem. Soc. Rev., 2013, 42, 366.

CALIX[4]ARENE-BASED PHOTORESPONSIVE NANOSYSTEMS FOR ANTIBACTERIAL APPLICATIONS




Andreotti G.a,Cimmaruta C.a,b,Monticelli M.a,b,Citro V.b, Cubellis M.V.b

aInstitute of Biomolecular Chemistry-CNR, via Campi Flegrei 34,

80078 Pozzuoli-Naples, Italy, bDepartment of Biology, Università degli Studi di Napoli Federico II,

via Cinthia, 21, 80126, Naples, Italy [email protected],

Fabry Disease (FD) is a rare pathology, caused by mutations in the gene GLA (located on the X chromosome) which encodes lysosomal alpha-galactosidase (AGAL) (1). Enzyme replacement (ERT) is the only approved specific therapy for FD. The large phenotypic spectrum of the disease highlights several problems and the cost-effectiveness of ERT is still debated especially for patients who have residual enzyme activity (2). ERT, which requires intravenous administration of the recombinant AGAL, is safe but it is expensive. For a correct analysis of its cost-effectiveness we should wait until a correct stratification of patients is possible.

Due to diffusion of screening projects among newborns and the general population, the number of patients harboring mild FD mutations will grow (3). For these patients it could be sufficient to enhance the residual activity of their mutant AGAL using low molecular weight chemicals which can act directly, binding and stabilizing the target mutant or indirectly, enhancing the synthesis of the enzyme, inhibiting its degradation, promoting protein folding etc. Pharmacological chaperones (PC) are those molecules that act directly. In FD, about 40% of all missense mutations are associated with a biochemically mildly damaged enzyme. A therapy with sub-inhibitory concentrations of 1-deoxygalactonojirimycin (DGJ), has been tested and very recently (April 2016) the EMA has recommended granting a marketing authorization in the EU for Galafold (migalastat) for the treatment of FD. Such therapy requires a fine-tuning of the dosage regimen because DGJ act as an inhibitor of AGAL, and it is suitable only for some patients.

The approach with small molecules for FD is promising, but DGJ is not yet the ideal drug, it should be considered as a first generation drug. We are looking for a drug with a more favorable balance between pharmacological chaperoning and enzymatic inhibition that would assure a larger therapeutic window and an improved therapeutic index. We can find either molecules that substitute DGJ or molecules that act in synergy with it, allowing a lower dosage. Acknowledgment: Research funded by Telethon-2012 (GGP12108) References:

4. Germain DP. 2010. Fabry disease. Orphanet J Rare Dis 5:30 5. Rombach SM et al. 2013. Cost-effectiveness of enzyme replacement therapy for Fabry disease. Orphanet

J Rare Dis 8:29. 6. Smid BE et al. 2015. Diagnostic dilemmas in Fabry disease: a case series study on GLA mutations of

unknown clinical significance. Clin Genet 88:161-166.

PHARMACOLOGICAL CHAPERONES TO CURE FABRY DISEASE



Cutignano A., Luongo E., Nuzzo G., Pagano D., Manzo E., Sardo A., Botte P., d’Ippolito G., Fontana A.

Institute of Biomolecular Chemistry, CNR, via Campi Flegrei 34,

80078 Pozzuoli (NA), Italy

[email protected]

Microalgal lipids are of great interest for their potential as food and feed ingredients, nutraceutical and cosmetic components, and as source of energy. Glycerol-based lipids constitute up to 50% or more of the biomass of microalgae. How synthesis of these products takes place is still poorly explored, although remarkable differences with plants and among microalgal species have already emerged. These differences attain to both lipid class and fatty acyl composition, which could be modulated by specific growth conditions. Here we present a new method based on UHPLC coupled to high resolution tandem mass spectrometry for targeted analysis of the main lipid classes, i.e. glycolipids, phospholipids and triacylglycerols integrated with a recently proposed MTBE-based protocol for lipid extraction and validated on five marine microalgal extracts exhibiting a great variety in lipid composition. The entire analytical workflow has been applied to investigate metabolic quali/quantitative changes of lipid pools in the diatom Cyclotella cryptica grown under different nutrient regimes.

A NOVEL UHPLC/Q-EXACTIVE MS METHOD FOR LIPIDOMIC ANALYSIS

OF MARINE MICROALGAE



Dessì A.1, Dallocchio R.1, Pani G. 2, Azara E.1, Scherm B.2, Migheli Q.2, Delogu G.1

1CNR- Istituto di Chimica Biomolecolare, Trav. La Crucca 3- 07100 Sassari, Italy 2UNISS-Dipartimento di Agraria, V.le Italia, 39- 07100 Sassari, Italy

[email protected]

Trichothecene mycotoxins are produced by filamentous fungi belonging to the Fusarium genus and are among the major causes of crop loss in cereals, furthermore, they may cause severe toxicosis in mammals when contaminated grain or their derivatives are ingested1. In our previous article2 a collection of natural and natural-like phenols and hydroxylated biphenyls was tested in vitro and significant trichothecene inhibitory activity and fungitoxic effect were detected on F. culmorum over a range of concentrations comprised between 0.25-1.5 mM.

Aiming to provide further insight into the understanding of structure-activity relationship of trichothecene inhibitors and TRI5, we have adopted an approach based on modelling techniques. A model of the trichodiene synthase (TRI5) of the wheat fungal pathogen and type-B trichothecene producer F. culmorum was developed based on homology modelling with the crystallized protein of F. sporotrichioides. Eight phenolic molecules were selected for their ability to inhibit trichothecene production and/or fungal vegetative growth in F. culmorum.

Experimental biological activity, molecular descriptors and interacting-structures obtained from computational analysis were compared. Besides the catalytic domain, three privileged sites (sites 1, 2 and 4) of the five identified on the protein surface, were evidenced. The TRI5-ligand interactions highlighted in this study represent a powerful tool to the identification of new Fusarium-targeted molecules with potential as trichothecene inhibitors.

References:

1. Bhat R, Rai RV, Karim AA. Compr. Rev. Food Sci. Food Saf. 2010, 9, 57-81. 2. Pani G., Scherm B., Azara E., Balmas V., Jahanshiri Z., Carta P., Fabbri D., Dettori M.A., Fadda A., Dessì

A., Dallocchio R., Migheli Q., Delogu G. J. Agric. Food Chem. 2014, 62, 4969-4978. 3. Pani, G., Dessì A., Dallocchio R., Scherm B., Azara E., Delogu G., Migheli Q. PLOSone 2016,

DOI:10.1371/journal.pone.0157316.

NATURAL PHENOLIC INHIBITORS OF TRICHOTHECENE BIOSYNTHESIS BY THE WHEAT FUNGAL PATHOGEN FUSARIUM CULMORUM: A

COMPUTATIONAL INSIGHT INTO THE STRUCTURE-ACTIVITY RELATIONSHIP



Dettori M.A. a, Fabbri D.a, Pani G.b, Balmas V.b, Migheli Q.b, Monti P.a,c, Rocchitta G.c, Serra P.A.c, and Delogu G.a

aCNR- Istituto di Chimica Biomolecolare, Trav. La Crucca 3- 07100 Sassari.

bUNISS-Dip. di Agraria, V.le Italia, 39- 07100 Sassari cUNISS- Dip. di Med. Clinica e Sperimentale, V.le S. Pietro, 43- 07100 Sassari.

[email protected]

We prepared a small collection of phenols and its C2-symmetry hydroxylated biphenyls to be applied for a novel nanotechnology-based approach in biosensor building and seed dressing. Biosensor technology provides easy-to-use and effective devices for medical, environmental and agro-food analyses. Amperometric biosensors are often coated with a polymeric permselective film, usually poly-Phenylendiamines (PD), to avoid electroactive interference by reducing agents, present in the target medium. We evaluated the permselectivity, stability and lifetime of polymers electrosynthesized from naturally occurring phenylpropanoids in monomeric and dimeric forms1. Among tested compounds, poly-magnolol and poly-isoeugenol can be regarded as effective and a healthier alternatives to poly-PD film in biosensor applications2.

The seed dressing (or coating) with conventional fungicides is a pest control technique against a broad range of species harmful to crops, and it allows fungi control with a reduced amount of fungicides during plant growth. Recently, we have selected and prepared a small collection of phenols and hydroxylated biphenyls effective against Fusarium culmorum in wheat crop3. In the present work we have investigate the effect of these phenols in the seed dressing by the only use of beta-cyclodextrin as biomatrix avoiding toxic compounds.

Our results indicate that a natural biphenol has the potential to protect wheat seeds from Fusarium damage in a manner that is both effective and environmentally acceptable. Application of sustainable seed dressing technology in organic agriculture may have significant commercial relevance.

References:

1. Calia, G.; Monti, P.; Marceddu, S.; Dettori, M.A.; Fabbri; D.; Jaoua, S.; O'Neill, R.D.; Serra, P.A.; Delogu, G. and Migheli Q. Analyst, 2015,140, 3607-3615.

2. Monti; P.; Calia, G.; Marceddu, S.; Dettori, M.A.; Fabbri, D.; Jaoua, S.; O'Neill, R.D.; Migheli, Q.; Delogu, G. and Serra P.A. Analyst, submitted.

3. Pani, G. Scherm, B.; Azara, E.; Balmas, V.; Jahanshiri, Z.; Carta, P.; Fabbri, D.; Dettori, M.A.; Fadda, A.; Dessì, A.; Dallocchio, R.; Migheli, Q. and Delogu, G. JAFC, 2014, 62, 4969.

PHENOLS AND HYDROXYLATED BIPHENYLS AS SUSTAINABLE COMPOUNDS FOR NANOTECHNOLOGIES


http://pubs.rsc.org/en/results?searchtext=Author%3AGiammario%20Calia

http://pubs.rsc.org/en/results?searchtext=Author%3APatrizia%20Monti

http://pubs.rsc.org/en/results?searchtext=Author%3ASalvatore%20Marceddu

http://pubs.rsc.org/en/results?searchtext=Author%3AMaria%20A.%20Dettori

http://pubs.rsc.org/en/results?searchtext=Author%3ADavide%20Fabbri

http://pubs.rsc.org/en/results?searchtext=Author%3ASamir%20Jaoua

http://pubs.rsc.org/en/results?searchtext=Author%3ARobert%20D.%20O'Neill

http://pubs.rsc.org/en/results?searchtext=Author%3APier%20A.%20Serra

http://pubs.rsc.org/en/results?searchtext=Author%3AGiovanna%20Delogu

http://pubs.rsc.org/en/results?searchtext=Author%3AQuirico%20Migheli

http://pubs.rsc.org/en/results?searchtext=Author%3APatrizia%20Monti

http://pubs.rsc.org/en/results?searchtext=Author%3AGiammario%20Calia

http://pubs.rsc.org/en/results?searchtext=Author%3ASalvatore%20Marceddu

http://pubs.rsc.org/en/results?searchtext=Author%3AMaria%20A.%20Dettori

http://pubs.rsc.org/en/results?searchtext=Author%3ADavide%20Fabbri

http://pubs.rsc.org/en/results?searchtext=Author%3ASamir%20Jaoua

http://pubs.rsc.org/en/results?searchtext=Author%3ARobert%20D.%20O'Neill

http://pubs.rsc.org/en/results?searchtext=Author%3AQuirico%20Migheli

http://pubs.rsc.org/en/results?searchtext=Author%3AGiovanna%20Delogu

http://pubs.rsc.org/en/results?searchtext=Author%3APier%20A.%20Serra



Di Mauro M.D.a,b, Tomasello B.b, Giardina R.C.b, Morrone R.a, Andronico, M.c, Renis M.b, D'Antona N.a

a CNR-Institute of Biomolecular Chemistry, via Gaifami 18, 95126, Catania

b Department of Drug Sciences, Biochemistry Section, University of Catania, viale Andrea Doria 6, 95125, Catania

c EtnaCosmesi, via 16a Traversa 60, 95032, Belpasso e-mail: [email protected]

The olive mill wastewater (OMWW), the main byproduct resulting from the olive oil extraction process, are annually produced in huge amount in olive-growing Mediterranean areas [1]. Although OMWW represent a serious environmental issue for their ecotoxicy and phytotoxicy, due to the high chemical and microbial content [2-3], they are still rich in different valuable compounds, such as sugars, mineral compounds and polyphenols. During last years several valorization strategies have been developed in order to recover these compounds [4]. In particular, sugars and minerals are some components of the natural moisturizing factor and act as humectants, so they are of great interest for the cosmetic industry [5,6].

Our research group recovered an extract containing high quantity of sugars and minerals from OMWW through a green adsorption/desorption process. We evaluated, in vitro, the impact of different concentrations of this extract for 6h and 24h on cell viability and genotoxicity of fibroblasts, respectively by MTT (3-[4,5-dimethyl(thiazol-2-yl)-3,5-diphenyltetrazolium bromide) assay and Comet assay. Our results highlight the non-toxic effect of the extract and allow us to suggest the appropriate concentration to be used for cosmetic formulation. References:

1. Deeb, A.A., Fayyad, M.K., Alawi, M.A. Chromatogr. Res. Int. 2012, 1-8. 2. Magdich, S., Jarboui, R., Rouina, B.B., Boukhris, M., Ammar, E. Sci. Total Environ. 2012, 430, 209-216 3. Justino, C.I., Pereira, R., Freitas, A.C., Rocha-Santos, T.A.P., Panteleitchouk, T.S.L., Duarte, A.C.

Ecotoxicology. 2012, 21(2), 615-629. 4. Dermeche, S., Nadour, M., Larroche, C., Moulti-Mati, F., Michaud, P. Process. Biochem. 2013, 48, 1532-

1552. 5. Verdier-Sévrain, S., Bonté, F. J. Cosmet. Dermatol. 2007, 6, 75-82. 6. Rodrigues, F., Pimentel, F.B., Oliveira, M.B.P.P. Ind. Crops Prod. 2015, 70, 116-124.

SUGARS AND MINERALS ENRICHED EXTRACT RECOVERED FROM OLIVE MILL WASTEWATER FOR COSMETIC APPLICATIONS

http://www.ncbi.nlm.nih.gov/pubmed/?term=Justino%20CI%5BAuthor%5D&cauthor=true&cauthor_uid=22042608

http://www.ncbi.nlm.nih.gov/pubmed/?term=Pereira%20R%5BAuthor%5D&cauthor=true&cauthor_uid=22042608

http://www.ncbi.nlm.nih.gov/pubmed/?term=Freitas%20AC%5BAuthor%5D&cauthor=true&cauthor_uid=22042608

http://www.ncbi.nlm.nih.gov/pubmed/?term=Rocha-Santos%20TA%5BAuthor%5D&cauthor=true&cauthor_uid=22042608

http://www.ncbi.nlm.nih.gov/pubmed/?term=Panteleitchouk%20TS%5BAuthor%5D&cauthor=true&cauthor_uid=22042608

http://www.ncbi.nlm.nih.gov/pubmed/?term=Duarte%20AC%5BAuthor%5D&cauthor=true&cauthor_uid=22042608

http://www.ncbi.nlm.nih.gov/pubmed/22042608



Fabbri D., Dettori M.A., Carta P. and Delogu G.

C.N.R. Institute of Biomolecular Chemistry - Traversa La Crucca 3-07100 Sassari.

[email protected]

1,3-Diaryl-2-propen-1-ones, commonly known as chalcones, have displayed remarkable variety of biological activities, among which anti-malarial, anti-inflammatory, immunomodulatory and antitumoral1. This class of anticancer agents have shown promising therapeutic efficacy for the management of human cancers. Chalcones, considered as precursors of flavonoids and isoflavonoids, are abundant in edible plants. In our laboratory we are actively engaged in the synthesis and biological evaluation (e.g. anti-malignant melanoma cancer) of hydroxylated natural-like biphenyls2. Hydroxylated biphenyl unit is embedded in many structures of bioactive natural products and some of them are present in compounds of high biological relevance. Compared to 2-methoxy phenols, 3,3’-dimethoxy-2,2’-dihydroxy-biphenyls generally manifest higher activity and less toxicity than the corresponding monomers. It is also known that the presence of a prenylated or unsaturated alkyl chain on flavonoids and chalcones, can lead to a remarkable increase in bioactivity3.

We herein report the synthesis, by Claisen–Schmidt condensation reaction base catalysed, of novel C2-symmetry hydroxylated (Type A and B) and prenylated chalcones based on a biphenyl scaffold and their corresponding monomers.

References:

1. Sharma, A. et al. Biorg. & Med. Chem. 2010, 18, 4711-4720. 2. Pisano, M. et al. BMC Cancer, 2016, 16(1):317 3. Tatsuzaki, J. et al. J Nat. Prod. 2006, 69, 1445–1449.

NEW SYNTHETIC HYDROXYLATED BIPHENYL CHALCONES, A POTENTIAL BIOACTIVE AGENTS FROM NATURAL SOURCES



Geraci C., Granata G., Consoli G.M.L.

Institute of Biomolecular Chemistry, National Research Council (CNR), Via Paolo Gaifami 18, 95126 Catania, Italy,

[email protected]

Due to increase of public interest in healthier and functional foods, the use of nanotechnological approaches to safeguard active ingredient is a challenge that has attracted the attention of researchers and food companies.1

In particular, the polymeric nanocapsules are an efficacy strategy to increase solubility and bioavailability of functional biomolecules in food, to protect from degradation and to mask unpleasant tastes.2

In this context, we report the synthesis of biocompatible and biodegradable polymeric nanocapsules (Figure 1) prepared by interfacial deposition of performed polymer method. The nanocapsules are able to load in lipid-core lipophilic molecules having pharmacological activities.3 Among the bioactive ingredients, we have focused our interest in hydroxycinnamic acids, presenting in the diet, which possess demonstrated chemoprevention properties.4,5 Light scattering techniques have been used to characterize and screen the prepared colloidal suspensions. Experimental data to describe the encapsulation process, as drug loading, encapsulation efficiency and time stability are given here. Preliminary tests performed to simulate gastro-intestinal digestion are also reported.

References:

1. Handford, C. E.; Dean, M.; Henchion, M.; Spence, M.; Elliot, C. T.; Campbell, K. Trends in Food Science &Technology 2014, 22, 226-241.

2. Ezhilarasi, P. N.; Karthik, P.; Chhanwal, N.; Anandharamkrishanan, C.; Food Bioprocess Technol. 2013,6, 628-647.

3. Venturini, C. G.; Jäger, E.; Oliveira, C. P.; Bernardi, A.; Battastini, A. M. O.; Guterres, S. S.; Pohlmann, A. R. Colloids Surf. A 2011, 375, 200-208.

4. Rocha, L. D.; Monteiro, M. C.; Teodoro, A. J. Cancer Clin. Oncol. 2012, 1, 109-121. 5. Rosa, L. S.; Silva, N. J. A.; Soares, N. C. P.; Monteiro, M. C.; Teodoro, A. J. J. Nutr. Food Sci. 2016, 6, 2-7.

POLYMERIC NANOCAPSULES LOADED WITH HYDROXYCINNAMIC ACIDS



Rainaldi G.1, Paris D.2, Melck D.2, Battaglia A.3, Fattorossi A.3, Ascierto P.A.4, Sini M.C.5, Palomba G.5, Rozzo C.5, Pisano M.5, Motta A.2, Palmieri G.5, Manca A.5

1Dipartimento di Ematologia, Oncologia e Medicina Molecolare,

Istituto Superiore di Sanità, Roma; 2Consiglio Nazionale Ricerche, Istituto di Chimica Biomolecolare, Pozzuoli, Napoli;

3Laboratorio di Immunologia, Polo scienze della salute della donna e del bambino, Fondazione Policlinico Universitario Agostino Gemelli, Roma;

4Istituto Nazionale Tumori Fondazione “Pascale”, Napoli; 5Consiglio Nazionale Ricerche, Istituto di Chimica Biomolecolare, Sassari

[email protected]

Background. Melanoma is an aggressive form of cancer difficult to treat because of its inherent resistance to standard treatments and its poor immunogenicity. Specific gene mutations, such as those in BRAF, NRAS and p16CDKN2A, could play a key role in radioresistance and immunogenicity of melanomas and be associated with specific metabolomic signatures whose identification could help define predictive markers. For this purpose three-dimensional multicellular spheroids, which mimic many aspects of solid tumors in vivo and are therefore a good model to study the characteristics of melanomas and its interactions with the immune system, might be useful. Methods. About 30 melanoma cell lines, derived from tumor biopsies excised from melanoma patients at different stages of the disease are available for the study. They have been characterized for pathogenetic mutations in major genes involved in melanomagenesis: BRAF, NRAS, CDKN2A, KIT, PTEN. Three-dimensional melanoma spheroids have been grown from three of the above cell lines: GR-Mel (BRAFwt/NRASwt), CN-Mel (BRAFwt/NRASmut), and WM-266 (BRAFmut/NRASwt). These cell lines have been selected for such studies considering the BRAF/NRAS status as the main molecular characteristic of melanoma classification. The 1H-MNR measurements will carried out on melanoma spheroids. The mutation patterns of these melanoma spheroids have been assessed by NGS-based screening. Preliminary Results. Three-dimensional melanoma spheroids have very diverse growth features compared to the same cell line grown as bidimensional monolayer, underlining the importance to use spheroid model in evaluating the response to anticancer treatments in a more realistic manner. Melanoma spheroids with different mutational status have peculiar cell growth dynamics, suggesting that specific mutational status may confer particular pathogenic features to melanoma cells. Evaluation of the metabolomic features is ongoing. Conclusions. Our studies will contribute in defining the irradiation conditions suitable to induce immunogenic cell death in radioresistant melanomas and characterizing the mechanisms involved, toward the development of new biomarkers predicting the response to the irradiation treatment.

GENETIC AND METABOLOMIC ANALYSES OF MELANOMA THREE-DIMENSIONAL MULTICELLULAR SPHEROIDS TOWARD THE

ASSESSMENT OF SPECIFIC SIGNATURES ASSOCIATED WITH TUMOR RADIOSENSITIVITY AND IMMUNOGENECITY




Napoli E., Ruberto G.

Institute of Biomolecular Chemistry – Italian National Research Council, Via P. Gaifami 18, 95126 Catania

[email protected] The genus Origanum (Mentheae tribe, Labiatae family) comprises about 70 species, subspecies, varieties and hybrids, characterized by considerable morphological and chemical diversity1. Within this genus, O. vulgare L. is the most widespread and known species. Oregano plant parts and biochemical extracts (herb, leaf, essential oil, etc.) are commonly used in the food industry as a spice2, in the production of alcoholic beverages and for their pharmacological properties, including antimicrobial3, antioxidant4, antithrombin and antihyperglycaemic activities.

In Sicily, oregano is the most common spontaneous aromatic plant and is a marked feature of the interior and hilly coastal landscape. Recently oregano, together with other aromatic plants, are becoming an interesting economic resource mainly for marginal areas. Due their main use as spice, the quality of the productions are related to the quantity and quality of their essential oils. This study was carried out with the scope of evaluate the annual differences on quantity and quality of essential oils during a three years period of three farms located in the vocational area of Aragona (Sicily). In the same time, differences on yields and quality during each season were evaluated sampling oregano 6-8 times every week starting from the first flowering period. Analyses were performed with a combination of GC-FID and GC-MS.

An evaluation of essential oils profiles during the three-years of production is reported correlating the analysis also to environmental conditions registered in the same period. A settlement of the qualitative and quantitative parameters of the essential oils collected in the aforesaid three-years period was observed. References:

1. Kintzios, S.E.. Profile of the multifaceted prince of the herbs. In: Kintzios, S.E. (Ed.), Oregano – The Genera Origanum and Lippia. Taylor & Francis,2002.

2. Mäkinen, S.M., Pääkkönen, K.K. Processing, effects and use of oregano and marjoram in foodstuffs and in food preparations. In: Kintzios, S.E. (Ed.), Oregano – The Genera Origanum and Lippia. Taylor & Francis, London, 2002.

3. Paparella, A., Taccogna, L., Aguzzi, I., Chaves-Lo´ pez, C., Serio, A., Marsilio, F., Suzzi, G. Flow cytometric assessment of the antimicrobial activity of essential oils against Listeria monocytogenes. Food Contr, 2008, 19, 1174–1182.

4. Fasseas, M.K., Mountzouris, K.C., Tarantilis, P.A., Polissiou, M., Zervas, G. Antioxidant activity in meat treated with oregano and sage essential oils. Food Chem., 2007, 106, 1188–1194.

POLIANNUAL AND SEASONAL EVALUATION OF SICILIAN ORIGANUM VULGARE ESSENTIAL OILS



Pisano M.1*, Mauri P.2*, Rossi R.2, Colombino M.1, Casula M., Rozzo C.1, Serra M.1, Manca A.5, Palmieri G.1

1Consiglio Nazionale delle Ricerche, Istituto di Chimica Biomolecolare, Sassari 2Consiglio Nazionale delle Ricerche, Istituto di Tecnologie Biomediche, Segrate, Milano *These authors contribute equally

[email protected] In the context of personalized medicine, an important role will be played by enabling "omics" technologies for molecular profiling, such as genomic profiling (using approaches of next-generation sequencing, NGS), transcriptomic (using microarrays or quantitative PCR), proteomics (using mass spectrometry methods) and metabolomic (by NMR assays). The combination of these methodologies on both in vitro tumor models and, particularly, in vivo tissue samples will allow to identify biomarkers for diagnosis (associated with various clinical and pathological aspects of the disease), prediction of response and/or resistance to therapies, and prognosis.

On this regard, our group has set itself to achieve the characterization of genomic (genetic mutations and polymorphisms) and proteomic profiles in different melanoma cell lines from excised lesions in patients at different stages of the disease (primary tumor; skin, lymph node, or visceral metastases). The main goal is the identification of differentially expressed proteins derived from the comparison with different genetic patterns in melanoma cells in order to identify the biochemical and cellular processes of initiation and progression of the disease and elucidate potential mechanisms involved in the development of the different melanoma genotypes.

For the mutation analysis, genomic DNA was isolated from melanoma cell lines, using standard methods. Specimens were analyzed for mutations in most common genes involved in melanoma pathogenesis with a specific Panel on the Ion Torrent platform (PGM

sequencer). All variants detected by NGS were confirmed through PCR-based Sanger sequencing

For the proteomic analysis, the melanoma cell lines have been analyzed by MudPIT approach. Data obtained from the MudPIT analysis in the different melanoma cell lines at various stages of the disease have been investigated by using specific identification software (SEQUEST Cluster and Bioworks). All samples were analyzed in duplicate, therefore, having available a substantial number of listings protein, it was possible to compare most of them. The comparison was performed using the MAProMA software which allows the preliminary assessment of differentially expressed proteins through specific algorithms (DAVE - Differential Average- and -Differential Confidence Index-DCI). The Dave parameter is indicative of the difference in relative amounts of the same proteins found in the two samples being analyzed; the DCI parameter provides the absolute value of the differential expression confidence. These two parameters are thus used to identify proteins with a different level of abundance, which is present in the two samples in a manner significantly different.

From a preliminary analysis of the data - we have just generated, a significant number of differentially expressed proteins was found to play an essential role in cancer tumorigenesis, including those involved in proliferation control, cellular motility, and resistance to apoptosis of melanoma cells. More accurate correlations and association analyses between genetic and proteomic data are ongoing.

PROTEOMIC ANALYSIS IN MELANOMA CELL LINES WITH DIFFERENT GENOTYPES



Poli A., Di Donato P., Tommonaro G., Nicolaus B.

Istituto di Chimica Biomolecolare, C.N.R., Via Campi Flegrei 34, 80078 Pozzuoli (Napoli)

[email protected]

Vegetable biomass is one of the most promising renewable source of energy and chemicals. Indeed, its exploitation for biofuel and commodity chemicals has been the focus of research over the last years and the gradual shift toward the so-called bio-based economy (an economy system based on exploitation of renewable resources) has determined a global increasing demand for goods mainly derived from vegetable biomass. In order to ensure a more sustainable productivity and the global food security, wastes coming from cultivation, harvesting and processing of vegetables have been recently considered as an alternative source for biomolecules and bioenergy that can find application in several industrial areas. According to this global scenario, the possible exploitation of the residual biomass coming from the industrial processing of lemon (Citrus limon), tomato (Lycopersicon esculentum var. ‘‘Hybrid Rome’’), fennel (Foeniculum vulgare var. dulce) and carrot (Daucus carota), were studied. This work highlighted the biorefinering approach for the re-use of such bio-agro-wastes that have been used as sources of two kinds of value added-products i.e polysaccharides and polyphenols, whose biotechnological properties and biological activities were investigated. Acknowledgments: This work was partially supported by the Project BioPoliS PON03PE_00107_1. References:

1. Poli A.; Finore I.; Tramice A.; Di Donato P.; Nicolaus B.; Lama L. 2016. Technical Developments for Vegetable Waste Biomass Degradation by Thermophiles, in Biotechnology of Extremophiles: Advances and Challenges, Editor Pabulo H. Rampelotto, DOI 10.1007/978-3-319-13521-2

2. Finore I.; Poli A.; Di Donato P.; Lama L.; Trincone A.; Fagnano M.; Mori M.; Nicolaus B.; Tramice A. Hemicellulose extract from Cynara cardunculus: a source of value-added biomolecules produced by xylanolytic thermozymes. Green Chemistry, 2016 doi: 10.1039/C5GC02774H

3. Di Donato P.; Taurisano V.; Tommonaro G.; Pasquale V.; Silvan J.M.; de Pascual S.T., Nicolaus B.; Poli A. 2016. Biological properties of polyphenols fractions recovered from fruits and vegetables’ industrial wastes. Submitted to Journal of Agricultural and Food Chemistry.

POLYPHENOLS AND POLYSACCHARIDES FROM VEGETABLE WASTES: BIOLOGICAL ACTIVITIES AND BIOTECHNOLOGICAL APPLICATIONS



Pozzebon A.a, Hussain R.b, Calderan A.a, Siligardi G.b and Ruzza P.a

aInstitute of Biomolecular Chemistry of CNR, Padua Unit, Padua, Italy.

bDiamond Light Source Ltd, Didcot, Oxfordshire, OX11 0DE, United [email protected]

Protein misfolding are associated with a growing number of neurodegenerative and peripheral disorders, characterized by the aggregation of different peptides and proteins into amyloid fibrils. Fibrils assembly is a complicated processes that involve formation of dimers and small oligomers, followed by growth into protofibrils and mature fibrils via a complex multi-step polymerization. Significant evidence indicates that a critical factor is represented by the destabilization of native conformation of proteins, which leads to protein aggregation; therefore, preventing this process may provide an effective therapeutic approach for treatment of amyloid-related diseases. Recently, we have reported on stabilization of native folded state of amylodogenic GFAP protein [1] as well as on inhibition of fibrillation of α-synuclein [2], using ceftriaxone (Cef), a safe and multi-potent β-lactam antibiotic.

Human lysozyme has been widely used over the past years as a model system to study many aspects of protein structure and function, including the mechanism of protein folding and the identification of protein stability determinants. In addition, five natural mutations in the human lysozyme gene have been reported, all located in the same region of the native structure of lysozyme, namely β-domain. All the mutations was found to be associated with systemic non-neuropathic amyloidosis, a disease in which the variant proteins misfold and form amyloid fibrils in the extracellular space of multiple organs and tissues.

In the present work, with the aim to elucidate the capability of ceftriaxone to interact with w.t and mutated human lysozyme, we synthesized the peptide corresponding to the sequence 69-82 of lysozyme, which has been demonstrated to have high propensity of fibril formation [3], and its I74T variant. SRCD and fluorescence spectroscopies have been used to obtain structural information about the peptide-Cef interactions and the effect of Cef on the peptide stability. Acknowledgment: We thank Diamond Light Source for access to beamline B23 (SM12591) 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:

1. Ruzza, P.; Vitale, R. M.; Hussain, R.; Biondi, B.; Amodeo, P.; Sechi, G. P.; Siligardi, G. Biochim. Biophys. Acta-Gen. Subj. 2016.

2. Ruzza, P.; Siligardi, G.;Hussain, R.; Marchiani, A.; Islami, M.; Bubacco, L.; Delogu, G.; Fabbri, D.; Dettori, A.M.; Sechi, M.; Nicolino Pala, N.; Spissu, Y.; Migheli, R.; Serra, P.A.; Sechi, G.P. ACS Chem. Neurosci. 2014, 5, 30-38.

3. Tokunaga, Y.; Matsumoto, M.; Sugimoto, Y. Int. J. Biol. Sci. 2015, 80, 208-216.

ANALYSIS OF THE INTERACTION WITH CEFTRIAXONE OF A PEPTIDE FROM THE CORE REGION OF HUMAN LYSOZYME



Villano R. a, Acocella M.R.b, Scettri A.b

aCNR-Istituto di Chimica Biomolecolare, Trav. La Crucca 3-07100 Sassari

bUniversità di Salerno-Dipartimento di Chimica e Biologia-Via Giovanni Paolo II, 132 - 84084 - Fisciano (SA)

[email protected]

In recent years, notable results have been achieved in the asymmetric direct aldol addition of ketones to aldehydes by employing secondary amines as organocatalysts.1 Although several functionalized ketones have been used as nucleophiles, the influence of a chiral group on the aromatic aldehydes on the reactivity is still poorly explored. The aldehyde 1, easily accessible in racemic and enantiomerically enriched form,2 seemed to be the ideal substrate to explore this aspect in the L-proline catalyzed aldol addition of acetone.

Interesting aspects, such as 1,4-asymmetric induction by the remote sulfinyl group of the aldehyde but also the stereochemical control and kinetic resolution of the racemic substrate by the chiral organocatalyst were examined. Finally, an unexpected stereoselective retro-aldol reaction catalyzed by L-proline was pointed out.3

Acknowledgment: Research funded by MIUR and University of Salerno References:

1. a) Modern Aldol Reactions (Ed.: R. Mahrwald), Wiley-VCH, Weinheim, 2004; b) Sustainable Catalysis: Without Metals or Other Endangered Elements (Ed.: M. North), Royal Society of Chemistry, 2016.

2. Massa, A.; Acocella, M. R.; De Sio, V.; Villano, R.; Scettri, A. Tetrahedron: Asymmetry 2009, 20, 202-204. 3. Villano, R.; Acocella, M. R.; Scettri, A. Tetrahedron 2016, in press, accepted manuscript.

INFLUENCE OF A REMOTE SULFINYL GROUP ON L-PROLINE-CATALYZED DIRECT ASYMMETRIC ALDOL ADDITION OF ACETONE



Vitale R.M. a, Piscitelli F.a, DI Marzo V.a, Amodeo P. a

a Instituto di Chimica Biomolecolare, Via Campi Flegrei 34,

80078, Pozzuoli (NA) [email protected]

The orphan GPR18 was considered the third cannabinoid receptor since the discovery of N-arachidonoyl glycine (NAGLY) as its putative endogenous agonist. However, discordant findings have been reported so far about the ability of GPR18 to respond to NAGLY as well as synthetic cannabinoids1. Recent reports1,2 suggest that the orphan receptor GPR18 is endowed with a high constitutive activity and, being overexpressed in melanoma metastases and inhibiting apoptosis, it could play an important role in tumor cell survival. Qin et al.2 ascribed GPR18 constitutive activity to the presence of an alanine residue at position 3.35 in place of the asparagine residue highly conserved among the class A of GPCR. This is in agreement with the constitutive activity of mutated forms of other receptors, such as CXCR4 and CXCR3, all bearing a N3.35A mutation. To demonstrate this hypothesis they obtained a GPR18 A3.35N variant exhibiting attenuated GPR18 signalling-mediated proliferation in histidine auxotrophic yeast. The absence of any structural data on this receptor prompted us to build a homology model of GPR18 to determine at molecular level the role of this non-conserved residue.

Sequence search against PDB using PSI-BLAST identified opioid receptors as the best hits. Multiple alignments were carried out and δ-opioid was finally chosen as structural template. The resulting model was then subjected to 200 ns molecular dynamics in a mixed lipid membrane/water environment containing a 30% molar concentration of cholesterol in a POPC bilayer. The choice of a cholesterol-containing membrane derived from the main localization of this receptor, like many other GPCRs, in lipid rafts. However, when GPCRs highly conserved residues in each TM helix were used to align the sequences, quite unexpectedly the position 3.35 corresponding to asparagine in the other GPCRs was indeed occupied by Y104 instead of the A108 residue proposed by Qin et al. In fact, A108 now occurs at position 3.39, taking as reference R3.50 in the DRY motif. The structural implications of the occurrence of a tyrosine at position 3.35 and a comparison between GPR18 A108N variant and wt are presented here. Acknowledgment: Research funded by GW Research Ltd

References:

1. Finlay DB, Joseph WR, Grimsey NL, Glass M. PeerJ. 2016, Mar 21;4:e1835. doi: 10.7717/peerj.1835.

eCollection 2016 2. Qin Y, Verdegaal EM, Siderius M, Bebelman JP, Smit MJ, Leurs R, Willemze R, Tensen CP, Osanto S.

Pigment Cell Melanoma Res. 2011 Feb;24(1):207-18

STRUCTURAL INSIGHTS INTO GPR18 FROM HOMOLOGY MODELING AND MOLECULAR DYNAMICS SIMULATIONS

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