title: Re-programming pullulan for targeting and controlled release of doxorubicin to the hepatocellular carcinoma cells.
abstract: A novel bioconjugate for hepatocellular carcinoma (HCC) targeting was obtained by pullulan re-programming, which involves the backbone oxidation and conjugation of targeting peptide and doxorubicin (Doxo) through a releasable linker. Preliminary in vivo studies showed that the oxidation of 40 glucopyranose units (GPU) out of 100 remarkably reduced the pullulan unspecific liver tropism. This oxidized polymer was functionalized with PreS1 to selectively target the HCC and with rhodamine (Rhod) as label to perform in vitro cell up-take investigations. PreS1 and Rhod were conjugated to the aldehydes present along the oxidized pullulan backbone through a 3.4 and 2kDa PEG spacer, respectively, and by reductive amination. The resulting PreS1-Pull-Rhod contained a mean of 8 PreS1 per oxidized pullulan chain. Cell culture studies were performed by using HepG2/SERPINB3 cells that overexpress the serpine B3 receptor and control HepG2/EMPTY cells that do not overexpress the receptor. A comparative study by cytofluorimetry and confocal microscopy performed using PreS1-Pull-Rhod and Pull-Rhod (control polymer) showed that PreS1 conveys to the conjugate high cell selectivity. Afterwards, the oxidized pullulan was exploited to generate a targeted drug delivery system by conjugation of Doxo to the polymer backbone through a hydrazone pH-sensitive bond and NH2-PEG3.4 kDa-PreS1. The PreS1-Pull-Doxo conjugate showed a two-fold increase of anticancer activity with respect to the control Pull-Doxo towards HepG2/SERPINB3 cells.
authors.affiliation: Department of Pharmaceutical & Pharmacological Sciences, University of Padova, Via F. Marzolo 5, 35131 Padova, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Balasso
authors.affiliation: Department of Pharmaceutical & Pharmacological Sciences, University of Padova, Via F. Marzolo 5, 35131 Padova, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Salmaso
authors.affiliation: Department of Medicine, University of Padova, via Giustiniani, 2, 35128 Padova, Italy.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Pontisso
authors.affiliation: Department of Surgery Oncology and Gastroenterology, University of Padova, via Giustiniani 2, 35124 Padova, Italy; Veneto Institute of Oncology IOV-IRCCS, via Gattamelata 64, 35128 Padova, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Rosato
authors.affiliation: Department of Medicine, University of Padova, via Giustiniani, 2, 35128 Padova, Italy.
authors.forename: Santina
authors.initials: S
authors.lastname: Quarta
authors.affiliation: Department of Pharmaceutical & Pharmacological Sciences, University of Padova, Via F. Marzolo 5, 35131 Padova, Italy.
authors.forename: Alessio
authors.initials: A
authors.lastname: Malfanti
authors.affiliation: Department of Pharmaceutical & Pharmacological Sciences, University of Padova, Via F. Marzolo 5, 35131 Padova, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Mastrotto
authors.affiliation: Department of Pharmaceutical & Pharmacological Sciences, University of Padova, Via F. Marzolo 5, 35131 Padova, Italy. Electronic address: paolo.caliceti@unipd.it.
authors.forename: Paolo
authors.initials: P
authors.lastname: Caliceti
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000493:pharmacokinetics / Q000494:pharmacology; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy*; D002470:Cell Survival; D003692:Delayed-Action Preparations; D004317:Doxorubicin / Q000737:chemistry* / Q000493:pharmacokinetics / Q000494:pharmacology; D005260:Female; D005936:Glucans / Q000737:chemistry*; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000188:drug therapy*; D008807:Mice, Inbred BALB C; D010084:Oxidation-Reduction; D011134:Polysaccharides / Q000737:chemistry / Q000494:pharmacology; D014018:Tissue Distribution
chemical_list: D000970:Antineoplastic Agents; D003692:Delayed-Action Preparations; D005936:Glucans; D011134:Polysaccharides; D004317:Doxorubicin; C009109:pullulan
keywords: Anticancer drug delivery; Hepatocellular carcinoma; Polymeric conjugates; Polysaccharides; Pullulan