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Nuovi pirrolochinolinoni angolari quali potenziali agenti fotochemioterapici: studio del meccanismo d’azione.
Introduzione:  Furocumarine 8-metossipsoralene (8-MOP) Angelicina Pirido[3,4-c]psoralene Benzo[4’,5’-d]psoralene Pirrolo[3,2,1-ij]chinolin-4-one 4,7-dimetilfuro[2,3-g]chinolin-2-one
Pirrolo[2,3- h ]chinolin-2-oni B-86 B-87 B-88 B-89 B-94 B-95 B-96 B-120 (lead compound)
Scopo della tesi ,[object Object],[object Object],[object Object],[object Object]
Caratteristiche spettroscopiche
Solubilità e coefficienti di ripartizione
Rese quantiche di fluorescenza B-86
Determinazione dell’attività antiproliferativa (HL-60)
Determinazione dell’attività antiproliferativa (HT-1080)
Determinazione dell’attività antiproliferativa (LoVo)
Effetto di  scavengers  sull’attività antiproliferativa   (B-86, B-88 e B-96) ,[object Object],[object Object]
Fotodegradazione dei composti (photobleaching)
Fotodegradazione (continua) ,[object Object],[object Object],[object Object]
Caratterizzazione dei fotoprodotti:  LC/TOF-MS (B-86)
LC-TOF/MS (B-86)
Spettri di assorbimento dei fotoprodotti
Conclusioni sulla fotodegradazione ,[object Object],[object Object],[object Object]
Produzione di specie reattive dell’ossigeno Produzione di  1 O 2   mediante RNO bleaching. Produzione di   O 2 -. attraverso il saggio dell’NBT.
Microscopia ad epifluorescenza *   Per capire dove i pirrolochinolinoni agissero si è effettuata una localizzazione intracallulare tramite microsc. ad epifl. B-86 B-95 B-96 U.V. Campo chiaro
Localizzazione intramitocondriale  tramite TMRM (B-88) B-88 TMRM  B-88 +  TMRM
Localizzazione intralisosomiale  tramite acridina orange (B-88) B-88 AO B-88 + AO
Interazione con il DNA ,[object Object]
Dicroismo lineare in flusso (continua) ,[object Object],[object Object]
Elettroforesi orizz. su gel di agarosio  (plasmide pBR322) Controllo B-86 B-88
Interazione con le proteine ,[object Object],Albumina
Elettroforesi verticale su gel di acrilammide  (fotodanno alle proteine di membrana) In condizioni aerobiche avviene la polimerizzazione proteica In condizioni anaerobiche non si assiste alla formazione di legami crociati
Quantificazione dei gel di acrilammide. ,[object Object]
Dicroismo circolare 0 20 Albumina RNAasi
Dicroismo circolare (continua) B-86 B-86 ,[object Object],[object Object],[object Object]
Interazione con i fosfolipidi ,[object Object]
Conclusioni ,[object Object],[object Object],[object Object]
[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Fotodegradazione B-96
*Caratterizzazione dei fotoprodotti1(MS)
*Caratterizzazione dei fotoprodotti2 Cinetiche di fotodegradazione di alcuni pirrolochinolinoni studiati.
Localizzazione cellulare dopo irradiazione U.V. B.F.
Fotoreazione sulla ribonucleasi A ,[object Object],[object Object],[object Object]

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