Pancreatic ductal adenocarcinoma is the fourth leading cause of oncological death in Italy and, due to its level of aggression, is set to become the second leading cause by 2030. A distinctive feature of this cancer is its abundant stroma formed by an extracellular matrix, stellate pancreatic cells and fibroblasts; which is capable of hindering access of common chemotherapeutic drugs to the cancer cells, therefore, there is an extreme need for selective and effective pharmacological approaches to counteract this disease.. The present work documents the synthesis of analogues of the known mitochondriotropic inhibitor of the potassium channel mitoKv1.3, PAPTP, as an innovative approach to fight the neoplasia and overcome the therapeutic limitations encountered by current chemotherapy and radiotherapy treatments. My thesis work was carried out in the laboratory and under the supervision of Prof. Andrea Mattarei (Department of Pharmaceutical Sciences, University of Padua). Specifically, four new compounds were synthesized by me through multi-step syntheses. The first two molecules are potential molecular probes to be applied as chemical tools in imaging experiments in cultured cells. The main purpose for which they were synthesized, is to confirm the mitochondrial localization of the PAPTP molecule, and possibly its interaction with the mitoKv1.3 channel in cultured cells, by means of fluorescence microscopy and transmission electron microscopy as imaging techniques.. The other two synthesized molecules are small organic compounds conjugated with the gastrin peptide. This peptide has proved to be a promising tool for selective drug delivery in the treatment of this cancer, being able to selectively bind to the extracellular receptors CCK1R and CCK2R, both overexpressed in the pancreatic ductal adenocarcinoma. The first of these conjugates will be used as a fluorescent probe for fluorescence microscopy imaging to assess the effective internalization of the peptide-drug chimera in cells overexpressing the gastrin receptor. The second conjugate bioreversibly links the gastrin peptide with the PAP-TP drug for the selective release of the drug in the pancreatic ductal adenocarcinoma in in vivo murine models of the pathology
L’adenocarcinoma duttale pancreatico rappresenta la quarta causa di morte oncologica in Italia e, in ragione del livello di aggressività, è destinato a diventare la seconda causa entro il 2030. Una caratteristica peculiare di questa neoplasia è il suo abbondante stroma, formato da una matrice extracellulare, cellule pancreatiche stellate e fibroblasti, che è in grado di ostacolare l’accesso ai comuni farmaci chemioterapici nel sito d’azione, per questo c’è un’estrema necessità di approcci farmacologici selettivi ed efficaci per contrastare questa patologia. Il presente lavoro documenta il processo sintetico per la preparazione di molecole analoghe del noto inibitore mitocondriotropico del canale del potassio mitoKv1.3, PAPTP, quale approccio innovativo per combattere la neoplasia e superare i limiti terapeutici che incontrano gli attuali trattamenti chemioterapici e radioterapici. Il mio lavoro di tesi è stato svolto presso il laboratorio e sotto la supervisione del Prof. Andrea Mattarei (Dipartimento di Scienze del Farmaco, Università degli studi di Padova). Nello specifico sono stati da me sintetizzati quattro nuovi composti attraverso sintesi a più passaggi. Le prime due molecole sono delle potenziali sonde molecolari da impiegare come strumenti chimici in esperimenti di imaging in colture cellulari. Lo scopo principale per il quale sono state sintetizzate è confermare la localizzazione a livello mitocondriale della molecola PAPTP e possibilmente la sua interazione con il canale mitoKv1.3 in cellule in coltura, usando come tecniche di imaging la microscopia di fluorescenza e la microscopia a trasmissione elettronica. Le altre due molecole sintetizzate sono delle piccole molecole organiche coniugate con il peptide della gastrina. Questo peptide si è rivelato essere un promettente strumento per il delivery selettivo di farmaci nel trattamento della neoplasia, essendo in grado di legarsi selettivamente ai recettori extracellulari CCK1R e CCK2R, entrambi sovraespressi nell’adenocarcinoma pancreatico duttale. Il primo di questi coniugati sarà utilizzato come sonda fluorescente per l’imaging attraverso la microscopia fluorescenza, allo scopo di valutare l’effettiva internalizzazione della chimera farmaco-peptide in cellule sovraesprimenti il recettore della gastrina. Il secondo coniugato lega il peptide della gastrina in maniera bioreversibile con il farmaco PAP-TP, per il rilascio selettivo del farmaco all’interno dell’adenocarcinoma duttale pancreatico in un modello murino della patologia .
Sintesi e caratterizzazione di strumenti chimici per l'imaging in sistemi biologici e di coniugati farmaco-peptide per la terapia mirata dell'adenocarcinoma duttale pancreatico
FOCHESATO, RICCARDO
2022/2023
Abstract
Pancreatic ductal adenocarcinoma is the fourth leading cause of oncological death in Italy and, due to its level of aggression, is set to become the second leading cause by 2030. A distinctive feature of this cancer is its abundant stroma formed by an extracellular matrix, stellate pancreatic cells and fibroblasts; which is capable of hindering access of common chemotherapeutic drugs to the cancer cells, therefore, there is an extreme need for selective and effective pharmacological approaches to counteract this disease.. The present work documents the synthesis of analogues of the known mitochondriotropic inhibitor of the potassium channel mitoKv1.3, PAPTP, as an innovative approach to fight the neoplasia and overcome the therapeutic limitations encountered by current chemotherapy and radiotherapy treatments. My thesis work was carried out in the laboratory and under the supervision of Prof. Andrea Mattarei (Department of Pharmaceutical Sciences, University of Padua). Specifically, four new compounds were synthesized by me through multi-step syntheses. The first two molecules are potential molecular probes to be applied as chemical tools in imaging experiments in cultured cells. The main purpose for which they were synthesized, is to confirm the mitochondrial localization of the PAPTP molecule, and possibly its interaction with the mitoKv1.3 channel in cultured cells, by means of fluorescence microscopy and transmission electron microscopy as imaging techniques.. The other two synthesized molecules are small organic compounds conjugated with the gastrin peptide. This peptide has proved to be a promising tool for selective drug delivery in the treatment of this cancer, being able to selectively bind to the extracellular receptors CCK1R and CCK2R, both overexpressed in the pancreatic ductal adenocarcinoma. The first of these conjugates will be used as a fluorescent probe for fluorescence microscopy imaging to assess the effective internalization of the peptide-drug chimera in cells overexpressing the gastrin receptor. The second conjugate bioreversibly links the gastrin peptide with the PAP-TP drug for the selective release of the drug in the pancreatic ductal adenocarcinoma in in vivo murine models of the pathologyFile | Dimensione | Formato | |
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https://hdl.handle.net/20.500.12608/61290