title: Cellular uptake of glucoheptoamidated poly(amidoamine) PAMAM G3 dendrimer with amide-conjugated biotin, a potential carrier of anticancer drugs.
abstract: In search for soluble derivatives of PAMAM dendrimers as potential carriers for hydrophobic drugs, the conjugates of PAMAM G3 with biotin, further converted into glycodendrimer with d-glucoheptono-1,4-lactone, were prepared. Polyamidoamine dendrimer (PAMAM) of third generation, G3 was functionalized with four biotin equivalents covalently attached to terminal amine nitrogens via amide bond G34B. The remaining 28 amine groups were blocked by glucoheptoamide substituents (gh) to give G34B28ghor with one fluorescein equivalent (attached by reaction of G34Bwith fluorescein isothiocyanate, FITC) via thiourea bond as FITC followed by exhaustive glucoheptoamidation to get G34B27gh1F. As a control the G3 substituted totally with 32 glucoheptoamide residues, G3ghand its fluorescein labeled analogue G331gh1Fwere synthesized. The glucoheptoamidation of PAMAM G0 dendrimer with glucoheptono-1,4-lactone was performed in order to fully characterize the1H NMR spectra of glucoheptoamidated PAMAM dendrimers and to control the derivatization of G3 with glucoheptono-1,4-lactone. Another two derivatives of G3, namely G34B28gh1F'and G332ghF', with ester bonded fluorescein were also obtained. Biological properties of obtained dendrimer conjugates were estimated in vitro with human cell lines: normal fibroblast (BJ) and two cancer glioblastoma (U-118 MG) and squamous carcinoma (SCC-15), including cytotoxicity by reduction of XTT and neutral red (NR) assays. Cellular uptake of dendrimer conjugates was evaluated with confocal microscopy. Obtained results confirmed, that biotinylated bioconjugates have always lower cytotoxicity and 3-4 times higher cellular uptake than non-biotinylated dendrimer conjugates in all cell lines. Comparison of various cell lines revealed different dose-dependent cell responses and the lower cytotoxicity of examined dendrimer conjugates for normal fibroblasts and squamous carcinoma, as compared with much higher cytotoxic effects seen in glioblastoma cell line. Synthetized multi-functional conjugate (G34B27gh1F) is a promising candidate as biocompatible vehicle for hydrophobic molecules used in anticancer therapy.
authors.affiliation: Faculty of Chemistry, Rzeszów University of Technology, 6 Powstańców Warszawy Ave, 35-959 Rzeszów, Poland. Electronic address: luram@prz.edu.pl.
authors.forename: Łukasz
authors.initials: Ł
authors.lastname: Uram
authors.affiliation: Faculty of Chemistry, Rzeszów University of Technology, 6 Powstańców Warszawy Ave, 35-959 Rzeszów, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Szuster
authors.affiliation: Faculty of Medical Sciences, Rzeszów University of Information Technology and Management, 2 Sucharskiego Str, 35-225 Rzeszów, Poland.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Filipowicz
authors.affiliation: Faculty of Chemistry, Rzeszów University of Technology, 6 Powstańców Warszawy Ave, 35-959 Rzeszów, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Zaręba
authors.affiliation: Department of Drug Technology and Biotechnology, Faculty of Chemistry, Warsaw University of Technology, 75 Koszykowa Str, 00-664 Warsaw, Poland.
authors.forename: Elżbieta
authors.initials: E
authors.lastname: Wałajtys-Rode
authors.affiliation: Centre for Innovative Research in Medical and Natural Sciences, Faculty of Medicine, University of Rzeszów, Warzywna 1a, 35-310 Rzeszów, Poland.
authors.forename: Stanisław
authors.initials: S
authors.lastname: Wołowiec
mesh_terms: D000577:Amides / Q000737:chemistry / Q000493:pharmacokinetics*; D000970:Antineoplastic Agents / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry / Q000493:pharmacokinetics*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D050091:Dendrimers / Q000737:chemistry / Q000493:pharmacokinetics*; D004305:Dose-Response Relationship, Drug; D004337:Drug Carriers / Q000737:chemistry / Q000493:pharmacokinetics; D006801:Humans; D015394:Molecular Structure; D013329:Structure-Activity Relationship
chemical_list: D000577:Amides; D000970:Antineoplastic Agents; D050091:Dendrimers; D004337:Drug Carriers; C104700:PAMAM Starburst; D001710:Biotin
keywords: Anticancer vehicle; Biotin bioconjugate; Cellular uptake; Confocal microscopy; In vitro; Normal and cancer cells; PAMAM G3 dendrimer; Synthesis; Toxicity; d-Glucoheptono-1,4-lactone