The mode of action of phloretin and its analogs on the permeability of natural membranes for neutral and charged molecules, such as urea, glucose and chloride has been characterized 25 years ago. In contrast to signal molecules with primary effects on transport systems of natural membranes, phloretin also affects model membranes, i.e., artificial membranes, which do not contain proteins. Since the dipole potential reducing effect of phloretin on mono- and bilayers has been found, it became clear that its primary effect must be a biophysical one: phloretin adsorbs to lipid layers and changes biophysical parameters of these layers. The aim of this work was the characterization of the interaction between the surface-active molecule phloretin and artificial lipid layers. We were able to describe structural and functional parameters of the model systems mono- and bilayer as functions of one or few variables. One of these parameters, the dipole potential, measured as a function of the aqueous phloretin concentration, allowed a critical examination of the Langmuir adsorption model that has been postulated for the interaction between phloretin and lipid layers. Surface pressure versus area per lipid molecule isotherms and surface (dipole) potential change versus area per lipid molecule isotherms, measured at lipid monolayers, allowed a structural description of the phloretin-lipid interaction: phloretin integrates into monolayers dependent on the surface pressure and the phase state of the lipid. Calorimetric measurements confirmed the integration of phloretin into membranes because of the strong decrease of the phase transition temperature, but they also showed that the cooperativity of phase transition is hardly affected, even at very high amounts of phloretin in the membrane. Obviously the interaction between phloretin and lipids is restricted to the head groups, an integration into the hydrocarbon layer is unlikely. 2H NMR measurements with spherical unilamellar vesicles of headgroup-deuterated lipid showed changed quadrupolar splittings indicating the interaction between phloretin and headgroups of the lipids. / Die Wirkung von Phloretin und seinen Analogen auf die Permeabilität von natürlichen Membranen für bestimmte ungeladene und geladene Moleküle wie z.B. Harnstoff, Glukose und Chlorid ist bereits vor 25 Jahren beschrieben worden. Im Gegensatz zu Signalmolekülen mit Primärwirkungen auf Transportsysteme natürlicher Membranen wirkt Phloretin auch auf Modellmembranen, d.h., künstliche, reine Lipidmembranen, die keine Proteine enthalten. Nach der Entdeckung des dipolpotential-reduzierenden Effekts von Phloretin auf Monolayer und Bilayer war klar, daß dessen primäre Wirkung biophysikalischer Natur sein mußte: Phloretin adsorbiert an Lipidschichten und verändert biophysikalische Parameter dieser Schichten. Ziel dieser Arbeit war es, die Wechselwirkungen des oberflächenaktiven Moleküls Phloretin mit künstlichen Lipidschichten näher zu charakterisieren. Strukturelle und funktionelle Parameter der Modellsysteme Mono- und Bilayer konnten in Abhängigkeit einer oder weniger Variablen verfolgt und beschrieben werden. Einer dieser Parameter, das Dipolpotential, gemessen als Funktion der Phloretinkonzentration in der wässrigen Phase, erlaubte eine kritische Betrachtung des in der Literatur postulierten Langmuirschen Adsorptionsverhaltens von Phloretin. Oberflächendruck-molekulare Fläche Isothermen sowie Oberflächenpotential (Dipolpotential)-molekulare Fläche Isothermen, ermittelt an Lipidmonoschichten, erlaubten eine strukturelle Beschreibung der Phloretin-Lipid Wechselwirkung: Phloretin integriert in Monoschichten, wobei dieser Effekt stark abhängig ist vom Filmdruck und vom Phasenzustand des Lipids. Kalorimetrische Messungen bestätigten die Integration von Phloretin in Membranen durch eine starke Abnahme der Phasenübergangstemperatur, sie zeigten aber auch, daß die Kooperativität der Lipidmoleküle nur wenig beeinträchtigt wird, selbst bei sehr großen Mengen von Phloretin in der Membran. Die Wechselwirkung von Phloretin mit Lipiden ist offensichtlich beschränkt auf die Kopfgruppen, eine Integration in die hydrophobe Kohlenwasserstoffphase findet wahrscheinlich nicht statt. 2H NMR Messungen an sphärischen, unilamellaren Vesikeln aus kopfgruppen-deuteriertem Lipid zeigten unter dem Einfluß von Phloretin eine veränderte Quadrupol-Aufspaltung, was die Wechselwirkung von Phloretin mit den Kopfgruppen der Lipide belegt.

Made available in DSpace on 2014-06-11T19:25:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-04-29Bitstream added on 2014-06-13T20:32:59Z : No. of bitstreams: 1 arima_aa_me_botib.pdf: 1075360 bytes, checksum: 9d02ce6ddf64780e885cc7e1816fa812 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A floretina (3-(4-hidroxifenil)-1-(2,4,6-trihidroxifenil)-1-propanona) é uma dihidrocalcona presente principalmente em maçãs que ultimamente vem sendo empregada para fins terapêuticos e cosméticos. Essa molécula tem demonstrado interagir com membranas modelo neutras e carregadas, alterando a hidratação, a qual é importante tanto para a estabilidade de bicamadas, como para suas permeabilidades seletivas. Neste trabalho, o efeito da floretina foi estudado em lipossomos (vesículas) negativamente carregados de DMPG (dimiristoilfosfatidilglicerol) a baixa força iônica (10 mM Hepes pH 7.4 + 2 mM NaCI). Os resultados foram comparados com os obtidos para a barbaloína [10-glicopiranosil-1,8-dihidroxi-3-(hidroximetil)-9(10H)-antracenona], - uma antraquinona glicosilada extraída de folhas de diferentes plantas do gênero Aloe -, porque essa biomolécula demonstrou ser capaz de interagir fortemente com vesículas de DMPG a baixa força iônica. As alterações dinâmicas e estruturais foram monitoradas pela técnica espectroscópica de ressonância paramagnética eletrônica (RPE) através da incorporação de marcadores de spin nas membranas lipídicas. Os dados foram analisados através de parâmetros medidos diretamente sobre os espectros e por meio de parâmetros simulados (soluções da equação estocástica de Liouville para um conjunto de spins magnéticos). Os resultados de RPE no intervalo entre 5 ºC e 10 ºC mostraram que a floretina aumenta sensivelmente a anisotropia tanto na superfície quanto no centro das bicamadas; enquanto que entre 15 ºC a 19 ºC, tanto a floretina como a barbaloína aumentam a mobilidade e a fluidez no centro da bicamada. Para altos valores de temperaturas (35 ºC a 45 ºC) a adição da floretina aparentemente não alterou a hidratação, mas mudou significantemente o empacotamento e a difusão molecular na região das cabeças polares; enquanto... / Phloretin (3-(4-Hydroxyphenyl)-1-(2,4,6-trihydroxyphenyl)-1-propanone) is a dihydrochalcone present mainly in apples, which lately has been used for medical and cosmetic purposes. It has been shown that phloretin interacts with neutral and charged model membranes, changing the hydration, which is important as for bilayer stability as for its permeability properties. In this work, the effect of phloretin was studied in negatively charged DMPG (dimyristoylphosphatidylglycerol) liposomes (vesicles) at low ionic strength (10mM Hepes pH 7.4 + 2 mM NaCI). The results were compared with the biomolecule barbaloin (10-glucopyranosyl-1,8-dihydroxy-3-(hydroxymethyl)-9(10H)-anthracenone), - a known active ingredient extracted from leaves of different Aloe plants -, because this glycosilated anthraquinone has showed to interact strongly with DMPG vesicles at low ionic strength. The dynamics and structural changes were monitored by ESR (electron spin resonance), using spin labels incorporated in the membranes. The data were analyzed by using parameters measured directly on the spectra and by simulated parameters (solutions of the stochastic Liouville equation for a set of magnetic spins). The ESR results for the interval between 5 ºC and 10 ºC showed that phloretin increases strongly the anisotropy on the surface as well as at the bilayer core, while between 15 ºC to 19 ºC, phloretin and barbaloin increase the mobility and the fluidity at the bilayer core. For high temperature (35 ºC to 45 ºC) the presence of phloretin apparently did not alter the hydration, but changed significantly the packing and the motional diffusion of the lipid polar heads; while barbaloin increased the hydration on the surface as well as the bilayer core, but produced great structural changes (“folds”) mainly at the end of the hydrocarbon... (Complete abstract click electronic access below)

Phloridzin, an antidiabetic and antineoplastic agent usually found in fruit trees, is a dihydrochalcone constituent that has a clinical/pharmaceutical significance as a sodiumglucose linked transport 2 (SGLT2) inhibitor. Phloridzin never experienced widespread clinical usage in the pharmaceutical market due to its side effects and poor bioavailability when compared to other antidiabetic therapeutics. The poor bioavailability is primarily attributed to the degradation of the glycosidic bond of the phloridzin, resulting in the formation of phloretin, the aglycone of phloridzin and glucose. While phloretin displays a reduced capacity of SGLT2 inhibition, this nutraceutical shows enhanced antineoplastic activity in comparison to phloridzin. Gold nanoparticles (AuNPs) have been explored in improving the bioavailability of many drugs and therefore we opt for gold nanoparticle mediated delivery of phloridzin and phloretin and exploration of their anticancer mechanism. In this study, we have synthesized phloridzin and phloretin conjugated gold nanoparticles (Phl-AuNP and Pht-AuNP) in a single-step, rapid, biofriendly processes. The synthesized AuNPs morphology and elemental composition was characterized via transmission electron microscopy, UV-Vis spectroscopy, scanning electron microscopyenergy dispersive x-ray spectroscopy, and thermogravimetric analysis. Assessment of the antineoplastic potency of the dihydrochalcone-conjugated AuNPs against cancerous cell lines was accomplished through monitoring via flow cytometry. We posit that the functionalization of these chalcones onto the gold nanoparticles’ surface has improved the pharmacokinetic profile of phloridzin and phloretin.

Resumo: A floretina (3-(4-hidroxifenil)-1-(2,4,6-trihidroxifenil)-1-propanona) é uma dihidrocalcona presente principalmente em maçãs que ultimamente vem sendo empregada para fins terapêuticos e cosméticos. Essa molécula tem demonstrado interagir com membranas modelo neutras e carregadas, alterando a hidratação, a qual é importante tanto para a estabilidade de bicamadas, como para suas permeabilidades seletivas. Neste trabalho, o efeito da floretina foi estudado em lipossomos (vesículas) negativamente carregados de DMPG (dimiristoilfosfatidilglicerol) a baixa força iônica (10 mM Hepes pH 7.4 + 2 mM NaCI). Os resultados foram comparados com os obtidos para a barbaloína [10-glicopiranosil-1,8-dihidroxi-3-(hidroximetil)-9(10H)-antracenona], - uma antraquinona glicosilada extraída de folhas de diferentes plantas do gênero Aloe -, porque essa biomolécula demonstrou ser capaz de interagir fortemente com vesículas de DMPG a baixa força iônica. As alterações dinâmicas e estruturais foram monitoradas pela técnica espectroscópica de ressonância paramagnética eletrônica (RPE) através da incorporação de marcadores de spin nas membranas lipídicas. Os dados foram analisados através de parâmetros medidos diretamente sobre os espectros e por meio de parâmetros simulados (soluções da equação estocástica de Liouville para um conjunto de spins magnéticos). Os resultados de RPE no intervalo entre 5 ºC e 10 ºC mostraram que a floretina aumenta sensivelmente a anisotropia tanto na superfície quanto no centro das bicamadas; enquanto que entre 15 ºC a 19 ºC, tanto a floretina como a barbaloína aumentam a mobilidade e a fluidez no centro da bicamada. Para altos valores de temperaturas (35 ºC a 45 ºC) a adição da floretina aparentemente não alterou a hidratação, mas mudou significantemente o empacotamento e a difusão molecular na região das cabeças polares; enquanto... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Phloretin (3-(4-Hydroxyphenyl)-1-(2,4,6-trihydroxyphenyl)-1-propanone) is a dihydrochalcone present mainly in apples, which lately has been used for medical and cosmetic purposes. It has been shown that phloretin interacts with neutral and charged model membranes, changing the hydration, which is important as for bilayer stability as for its permeability properties. In this work, the effect of phloretin was studied in negatively charged DMPG (dimyristoylphosphatidylglycerol) liposomes (vesicles) at low ionic strength (10mM Hepes pH 7.4 + 2 mM NaCI). The results were compared with the biomolecule barbaloin (10-glucopyranosyl-1,8-dihydroxy-3-(hydroxymethyl)-9(10H)-anthracenone), - a known active ingredient extracted from leaves of different Aloe plants -, because this glycosilated anthraquinone has showed to interact strongly with DMPG vesicles at low ionic strength. The dynamics and structural changes were monitored by ESR (electron spin resonance), using spin labels incorporated in the membranes. The data were analyzed by using parameters measured directly on the spectra and by simulated parameters (solutions of the stochastic Liouville equation for a set of magnetic spins). The ESR results for the interval between 5 ºC and 10 ºC showed that phloretin increases strongly the anisotropy on the surface as well as at the bilayer core, while between 15 ºC to 19 ºC, phloretin and barbaloin increase the mobility and the fluidity at the bilayer core. For high temperature (35 ºC to 45 ºC) the presence of phloretin apparently did not alter the hydration, but changed significantly the packing and the motional diffusion of the lipid polar heads; while barbaloin increased the hydration on the surface as well as the bilayer core, but produced great structural changes ("folds") mainly at the end of the hydrocarbon... (Complete abstract click electronic access below) / Orientador: André Sampaio Pupo / Coorientador: Roberto Morato Fernandez / Banca: Carlos Frederico de Oliveira Graeff / Banca: Joel Meza Hormaza / Mestre