Dendrimers are multivalent, well-defined materials that constitute a new class of polymer macromolecules. They have been extensively studied over the past several decades mainly due to their exceptional structure properties. The size of molecule, number of terminal groups, molecular weight and several other properties of dendrimers could be precisely controlled during synthesis. The main goal of this thesis is to investigate the optical properties and light induced photochemical reaction dynamics within the PPI and PAMAM dendrimers functionalized by CAzPA and ESA type terminal groups by means of several spectroscopic techniques. Because photochromism of CAzPA and ESA type terminal groups is associated with light induced isomerization and tautomerization photochemical reactions, the main attention of present thesis was devoted to investigate dynamical properties of functionalized dendrimers as an influence of dendrimer type and its generation. Investigations of dynamic properties of PPI and PAMAM dendrimers functionalized with the CAzPA terminal groups have shown that isomerization rate is insensitive to dendrimer type, solvent and excitation wavelength. The isomerization rate of the dendrimer films take place in about 15 ps and it is only seven times slower than in dendrimer solvent. Both experimental and theoretical optical studies performed on PPI dendrimers functionalized with the ESA type terminal groups reveal four stable tautomeric forms with different energy in the... [to full text] / Dendrimerai tai naujai dendritinių polimerų klasei priskiriamos makromolekulės. Jų dydis, funkcinių grupių skaičius yra tiksliai apibrėžti ir gali būti kontroliuojami sintezės metu. Dėl išskirtinių struktūrinių savybių, dendrimerai jau keletą dešimtmečių yra intensyviai tiriamos medžiagos. Pagrindinis disertacijos tikslas yra ištirti PPI ir PAMAM dendrimerų, funkcionalizuotų CAzPA ir ESA fotochrominiais junginiais optines savybes bei šviesa inicijuotų fotocheminių reakcijų dinamiką panaudojus kelias tyrimų metodikas. Kadangi CAzPA ir ESA junginių fotochromizmas yra susijęs su šviesa indukuota izomerizacijos bei tautomerizacijos fotocheminėmis reakcijomis, todėl darbe buvo siekiama ištirti šiais junginiais funkcionalizuotų dendrimerų dinaminių savybių priklausomybę nuo dendrimero tipo bei jo generacijos. Ištyrus PPI ir PAMAM dendrimerų funkcionalizuotų CAzPA junginiais plėvelių ir tirpalų dinamines savybes, buvo nustatyta, kad izomerizacijos sparta nepriklauso nuo dendrimero tipo, tirpiklio ir žadinančios spinduliuotės energijos. Dendrimerų plėvelių sužadintos būsenos relaksacijos trukmė yra apie 15 ps, o tai apie 7 kartus lėčiau nei tirpaluose. Eksperimentiškai ir teoriškai ištyrus skirtingų generacijų PPI dendrimerų funkcionalizuotų ESA fotochrominiais junginiais optines savybes, buvo nustatytos keturios stabilios ESA funkcinių grupių tautomerinės formos, kurių pagrindinės būsenos energijos yra skirtingos. Skirtingų dendrimero generacijų sužadintos būsenos dinamikos... [toliau žr. visą tekstą]

Delivery of siRNAs for the treatment of tumors critically depends on the development of efficient nucleic acid carrier systems. The complexation of dendritic polymers (dendrimers) results in nanoparticles, called dendriplexes, that protect siRNA from degradation and mediate non-specific cellular uptake of siRNA. However, large siRNA doses are required for in vivo use due to accumulation of the nanoparticles in sinks such as the lung, liver, and spleen. This suggests the exploration of targeted nanoparticles for enhancing tumor cell specificity and achieving higher siRNA levels in tumors. In this work, we report on the targeted delivery of a therapeutic siRNA specific for BIRC5/Survivin in vitro and in vivo to tumor cells expressing the surface marker prostate stem cell antigen (PSCA). For this, polyplexes consisting of single-chain antibody fragments specific for PSCA conjugated to siRNA/maltose-modified poly(propylene imine) dendriplexes were used. These polyplexes were endocytosed by PSCA-positive 293TPSCA/ffLuc and PC3PSCA cells and caused knockdown of reporter gene firefly luciferase and Survivin expression, respectively. In a therapeutic study in PC3PSCA xenograft-bearing mice, significant anti-tumor effects were observed upon systemic administration of the targeted polyplexes. This indicates superior anti-tumor efficacy when employing targeted delivery of Survivin-specific siRNA, based on the additive effects of siRNA-mediated Survivin knockdown in combination with scFv-mediated PSCA inhibition. Among non-viral vectors, cationic polymers, such as poly(propylene imine) (PPI), play also a prominent role in plasmid DNA delivery. However, limitations of polycationic polymer-based DNA delivery systems are (i) insufficient target specificity, (ii) unsatisfactory transgene expression, and (iii) undesired transfer of therapeutic DNA into non-target cells. We developed single-chain antibody fragment (scFv)-directed hybrid polyplexes for targeted gene therapy of prostate stem cell antigen (PSCA)-positive tumors. Besides mono-biotinylated PSCA-specific single-chain antibodies (scFv(AM1-P-BAP)) conjugated to neutravidin, the hybrid polyplexes comprise β cyclodextrin-modified PPI as well as biotin/maltose-modified PPI as carriers for minicircle DNAs encoding for Sleeping Beauty transposase and a transposon encoding the gene of interest. The PSCA-specific hybrid polyplexes efficiently delivered a GFP gene in PSCA-positive tumor cells, whereas control hybrid polyplexes showed low gene transfer efficiency. In an experimental gene therapy approach, targeted transposition of a codon-optimized p53 into p53 deficient HCT116p53-/-/PSCA cells demonstrated decreased clonogenic survival when compared to mock controls. Noteworthily, p53 transposition in PTEN-deficient H4PSCA glioma cells caused nearly complete loss of clonogenic survival. These results demonstrate the feasibility of combining tumor-targeting hybrid polyplexes and Sleeping Beauty gene transposition, which, due to the modular design, can be extended to other target genes and tumor entities.