Design, synthesis, and characterization of new fluorescent probes for in vivo redox visualization
We have initiated a research program intended to prepare lipid soluble free radical sensitive fluorescent probes which can readily report the antioxidant activity within the lipid bilayers. This work provides a substantial amount of information useful in the rational design of pyrromethane dye-based...
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McGill University
2008
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Chemistry - Biochemistry |
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Chemistry - Biochemistry Oleynik, Paul Design, synthesis, and characterization of new fluorescent probes for in vivo redox visualization |
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We have initiated a research program intended to prepare lipid soluble free radical sensitive fluorescent probes which can readily report the antioxidant activity within the lipid bilayers. This work provides a substantial amount of information useful in the rational design of pyrromethane dye-based off-on fluorescent probes/switches for redox visualization. In this thesis we report the photophysical properties of a newly prepared Trolox®-PM605 dye (B-TOH) which exhibits a marked off-on fluorescence behaviour in the presence of radicals in homogeneous and microheterogeneous solutions. The absorption and fluorescence spectra, fluorescence quantum yields, and fluorescence lifetimes for this probe when dissolved in organic solvents, as well as in dimirystoyl phosphatidylcholine (DMPC) lipid vesicle suspensions in water are reported herein. The analogous photophysical properties were measured for PM605, the precursor fluorophore used in B-TOH synthesis, its deacylated derivative PM-OH, and a control probe 3,5-Di-tert-butyl-4-hydroxybenzoic acid-PM605 (PM-BHB, which is expected to have a lower reactivity towards radicals) adduct whose synthesis is further described herein. We also report the emission time profile of B-TOH, PM-BHB, and PM-OH following their prolonged exposure to peroxyl radicals in organic solvents and liposome water suspensions. Via careful electrochemical analysis and HPLC oxidation product studies, the observed emission growth of B-TOH is shown to occur via the deactivation of a Photoinduced Electron Transfer (PET) process, operative only in reduced B-TOH. Following exposure of B-TOH to free radicals and subsequent oxidation, PET is surpressed. In summary, we report a novel hydrophobic fluorescent antioxidant indicator with optimum off/on ratio properties for homogeneous solution studies. Antioxidant depletion and the onset of radical mediated oxidation can be directly monitored following emission enhancement over time, thus dramatically facilitati === Nous avons initié un programme de recherche qui a pour but de développer des sondes fluorescentes, sensibles aux radicaux libres, qui soient solubles dans les solutions lipidiques. Ces sondes devront pouvoir rapporter les activités anti-oxidantes des bi-couches lipidiques. Ce travail fournit des informations importantes pour la conception rationnelle de colorants dérivés du pyrrométhane qui pourront servir d'indicateurs fluorescents "on-off" de la visualisation des réactions redox. Dans cette thèse, nous rapportons les propriétés du nouvel indicateur Trolox-PM605 (B-TOH) qui montre un comportement fluorescent "on-off" marqué en présence de radicaux en solutions homogènes et micro-homogènes. Les spectres d'absorption et de fluorescence, les taux de fluorescence quantique, et la durée de vie de la fluorescence pour cette sonde quand dissoute dans des solvants organiques ainsi que dans des vésicules lipidiques de dimirystoyl de phoshatidylcholine (DMPC) en suspensions dans l'eau sont également rapportées. Les propriétés analogues photophysiques du PM605 (le précurseur fluorophore utilisé pour la synthèse du B-TOH) et du PM-OH (son dérivé acétyle) ainsi que celles d'une sonde contrôle, le 3,5-Di-tert-butyl-4-hydroxybenzoic acid-PM605 (PM-BHB, dont il est attendu d'observer une plus faible réactivité envers les radicaux libres) ont ainsi été mesurées. Nous rapportons également le profil temporel d'émission du B-TOH, du PM-BHB ainsi que du PM-OH, à la suite de leur exposition prolongée aux radicaux peroxydes en solution dans des solvants organiques ou des suspensions aqueuses de liposomes. De précises analyses électrochimiques et des études des produits d'oxidation HPLC ont permis l'observation de la croissance de l'émission du B-TOH, qui se déroule via la désactivation d'un processus de Transfert d'Electron Photo-induit (TEP), ce processus ne s'opérant que dans le contexte du B-TOH réduit. A la suite de l'exposition du B-T |
author2 |
Gonzalo Cosa (Internal/Supervisor) |
author_facet |
Gonzalo Cosa (Internal/Supervisor) Oleynik, Paul |
author |
Oleynik, Paul |
author_sort |
Oleynik, Paul |
title |
Design, synthesis, and characterization of new fluorescent probes for in vivo redox visualization |
title_short |
Design, synthesis, and characterization of new fluorescent probes for in vivo redox visualization |
title_full |
Design, synthesis, and characterization of new fluorescent probes for in vivo redox visualization |
title_fullStr |
Design, synthesis, and characterization of new fluorescent probes for in vivo redox visualization |
title_full_unstemmed |
Design, synthesis, and characterization of new fluorescent probes for in vivo redox visualization |
title_sort |
design, synthesis, and characterization of new fluorescent probes for in vivo redox visualization |
publisher |
McGill University |
publishDate |
2008 |
url |
http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=18789 |
work_keys_str_mv |
AT oleynikpaul designsynthesisandcharacterizationofnewfluorescentprobesforinvivoredoxvisualization |
_version_ |
1716637429320908800 |
spelling |
ndltd-LACETR-oai-collectionscanada.gc.ca-QMM.187892014-02-13T03:42:04ZDesign, synthesis, and characterization of new fluorescent probes for in vivo redox visualizationOleynik, PaulChemistry - BiochemistryWe have initiated a research program intended to prepare lipid soluble free radical sensitive fluorescent probes which can readily report the antioxidant activity within the lipid bilayers. This work provides a substantial amount of information useful in the rational design of pyrromethane dye-based off-on fluorescent probes/switches for redox visualization. In this thesis we report the photophysical properties of a newly prepared Trolox®-PM605 dye (B-TOH) which exhibits a marked off-on fluorescence behaviour in the presence of radicals in homogeneous and microheterogeneous solutions. The absorption and fluorescence spectra, fluorescence quantum yields, and fluorescence lifetimes for this probe when dissolved in organic solvents, as well as in dimirystoyl phosphatidylcholine (DMPC) lipid vesicle suspensions in water are reported herein. The analogous photophysical properties were measured for PM605, the precursor fluorophore used in B-TOH synthesis, its deacylated derivative PM-OH, and a control probe 3,5-Di-tert-butyl-4-hydroxybenzoic acid-PM605 (PM-BHB, which is expected to have a lower reactivity towards radicals) adduct whose synthesis is further described herein. We also report the emission time profile of B-TOH, PM-BHB, and PM-OH following their prolonged exposure to peroxyl radicals in organic solvents and liposome water suspensions. Via careful electrochemical analysis and HPLC oxidation product studies, the observed emission growth of B-TOH is shown to occur via the deactivation of a Photoinduced Electron Transfer (PET) process, operative only in reduced B-TOH. Following exposure of B-TOH to free radicals and subsequent oxidation, PET is surpressed. In summary, we report a novel hydrophobic fluorescent antioxidant indicator with optimum off/on ratio properties for homogeneous solution studies. Antioxidant depletion and the onset of radical mediated oxidation can be directly monitored following emission enhancement over time, thus dramatically facilitatiNous avons initié un programme de recherche qui a pour but de développer des sondes fluorescentes, sensibles aux radicaux libres, qui soient solubles dans les solutions lipidiques. Ces sondes devront pouvoir rapporter les activités anti-oxidantes des bi-couches lipidiques. Ce travail fournit des informations importantes pour la conception rationnelle de colorants dérivés du pyrrométhane qui pourront servir d'indicateurs fluorescents "on-off" de la visualisation des réactions redox. Dans cette thèse, nous rapportons les propriétés du nouvel indicateur Trolox-PM605 (B-TOH) qui montre un comportement fluorescent "on-off" marqué en présence de radicaux en solutions homogènes et micro-homogènes. Les spectres d'absorption et de fluorescence, les taux de fluorescence quantique, et la durée de vie de la fluorescence pour cette sonde quand dissoute dans des solvants organiques ainsi que dans des vésicules lipidiques de dimirystoyl de phoshatidylcholine (DMPC) en suspensions dans l'eau sont également rapportées. Les propriétés analogues photophysiques du PM605 (le précurseur fluorophore utilisé pour la synthèse du B-TOH) et du PM-OH (son dérivé acétyle) ainsi que celles d'une sonde contrôle, le 3,5-Di-tert-butyl-4-hydroxybenzoic acid-PM605 (PM-BHB, dont il est attendu d'observer une plus faible réactivité envers les radicaux libres) ont ainsi été mesurées. Nous rapportons également le profil temporel d'émission du B-TOH, du PM-BHB ainsi que du PM-OH, à la suite de leur exposition prolongée aux radicaux peroxydes en solution dans des solvants organiques ou des suspensions aqueuses de liposomes. De précises analyses électrochimiques et des études des produits d'oxidation HPLC ont permis l'observation de la croissance de l'émission du B-TOH, qui se déroule via la désactivation d'un processus de Transfert d'Electron Photo-induit (TEP), ce processus ne s'opérant que dans le contexte du B-TOH réduit. A la suite de l'exposition du B-TMcGill UniversityGonzalo Cosa (Internal/Supervisor)2008Electronic Thesis or Dissertationapplication/pdfenElectronically-submitted theses.All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated.Master of Science (Department of Chemistry) http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=18789 |