RESUMO
The spectroscopy and photophysics of the photosensitizer hypericin when in homogeneous solutions and when bound to liposomes were studied. Hypericin was found to partition efficiently into DMPC liposomes, with a binding constant of 58 (mg lipid/mL)-1. In these liposomes the singlet oxygen production quantum yield was 0.43 +/- 0.09. To determine the deactivation constant of singlet oxygen in lipid bilayers for the first time, we calculated extrapolated values from its quenching by DMPC and lecithin in homogeneous solutions and obtained decay times of 36.4 and 12.2 microseconds, respectively. We also measured the quenching of singlet oxygen, sensitized by hypericin in DMPC liposomes, by NaN3, diphenyl isobenzofuran and H2O:D2O mixtures and explained the results on the basis of singlet oxygen diffusing rapidly out of the lipid bilayer into the aqueous medium. The observed temperature effect on the lifetime of singlet oxygen of about 50% over a 15 degrees C range in liposome suspension contrasts with a 3% change in a homogeneous solution in 1-nonanol and is explained by the temperature effect on the diffusion out of the liposome. A strong pH effect was observed, indicating that the deprotonated species formed above about pH 10 is a much weaker photosensitizer of singlet oxygen than the native, protonated species.
Assuntos
Oxigênio/química , Perileno/análogos & derivados , Fármacos Fotossensibilizantes/química , Antracenos , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Cinética , Lipossomos , Oxigênio/efeitos da radiação , Perileno/química , Fotoquímica , Oxigênio Singlete , Soluções , EspectrofotometriaRESUMO
Tris (2,2'-bipyridyl)ruthenium(II)chloride hexahydrate (Ru[bpy]3(2+)) free in solution and adsorbed onto antimony-doped SnO2 colloidal particles was used as a photosensitizer for a comparison of the O2(1 delta g) and electron-transfer-mediated photooxidation of tryptophan (TRP), respectively. Quenching of excited Ru(bpy)3(2+) by O2(3 sigma g-) in an aerated aqueous solution leads only to the formation of O2(1 delta g) (phi delta = 0.18) and this compound was used as a type II photosensitizer. Excitation of Ru(bpy)3(2+) adsorbed onto Sb/SnO2 results in a fast injection of an electron into the conduction band of the semiconductor and accordingly to the formation of Ru(bpy)3(2+) and was used for the sensitization of the electron-transfer-mediated photooxidation. The Ru(bpy)3(3+) is reduced by TRP with a bimolecular rate constant kQ = 5.9 x 10(8) M-1 s-1, while O2(1 delta g) is quenched by TRP with kt = 7.1 x 10(7) M-1 s-1 (chemical + physical quenching). Relative rate constants for the photooxidation of TRP (kc) via both pathways were determined using fluorescence emission spectroscopy. With Np, the rate of photons absorbed, being constant for both pathways we obtained kc = (372/Np) M-1 s-1 for the O2(1 delta g) pathway and kc > or = (25,013/Np) M-1 s-1 for the electron-transfer pathway, respectively. Thus the photooxidation of Trp is more than two orders of magnitude more efficient when it is initiated by electron transfer than when initiated by O2(1 delta g).
Assuntos
Triptofano/química , 2,2'-Dipiridil/análogos & derivados , Transporte de Elétrons , Corantes Fluorescentes , Cinética , Cinurenina/análogos & derivados , Compostos Organometálicos , Fotoquímica , FotóliseRESUMO
Troglitazone (CS-045) is a new oral antidiabetic drug reported to be effective in insulin-resistant diabetes and to show antihypertensive effects. Photooxidation of troglitazone gave the quinone and quinone epoxide as the major final stable products. An intermediate observed by NMR spectroscopy was shown to be the hydroperoxydienone, which is moderately stable at room temperature. The rate constant of singlet oxygen quenching by troglitazone is 2.14 x 10(8) M(-1) s(-1) and the reaction rate constant in acetone-d6 is 8.64 X 10(6) M(-1) s(-1). Only the chroman ring of troglitazone reacts with and quenches singlet oxygen significantly, and its reactivity and products are analogous to those of alpha-tocopherol. The reactivity of CS-45 toward singlet oxygen is much larger than that of the related compounds lacking the chroman ring.
Assuntos
Cromanos/efeitos da radiação , Hipoglicemiantes/efeitos da radiação , Tiazóis/efeitos da radiação , Tiazolidinedionas , Cromanos/química , Estabilidade de Medicamentos , Hipoglicemiantes/química , Cinética , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Oxirredução , Oxigênio , Fotoquímica , Quinonas , Oxigênio Singlete , Relação Estrutura-Atividade , Tiazóis/química , TroglitazonaRESUMO
Epidemiologic studies strongly associate eosinophilia-myalgia syndrome (EMS) with ingestion of tryptophan containing a contaminant ("peak E"). Prior reports have suggested that peak E is the di-tryptophan N alpha-animal of acetaldehyde. Spectral and chemical studies now demonstrate that peak E is 1,1'-ethylidenebis[tryptophan]. This novel amino acid may be the etiological agent responsible for EMS, or it may be a marker of a still unidentified causal agent.
Assuntos
Eosinofilia/induzido quimicamente , Músculos , Dor/induzido quimicamente , Triptofano/análogos & derivados , Triptofano/toxicidade , Cromatografia Líquida de Alta Pressão , Humanos , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Espectrometria de Massas de Bombardeamento Rápido de Átomos , Síndrome , Triptofano/isolamento & purificaçãoRESUMO
The photooxidation of the antipsoriatic drug anthralin (1,8-dihydroxy-9-anthrone) has been studied by several kinetic techniques, including direct observation of 1O2 (1 delta g) luminescence at 1.27 microns. The rate of deactivation of 1O2 increases at higher pH, demonstrating that the trihydroxyanthracene anion is the reactive species. Direct determination of the rate constant of 1O2 deactivation (kR + kQ) in deuterated buffer systems by luminescence quenching gave a value of 3.0 x 10(8) M-1 s-1 for the anion; the neutral anthrone is unreactive. The rate constant for the neutral anthrone in benzene-d6 is 2.8 x 10(4) M-1 s-1. Competition experiments with tetramethylethylene in acetonitrile gave a rate constant for reaction alone (kR) of 2.1 x 10(8) M-1 s-1 for the anion.
Assuntos
Antralina/química , Cinética , Luminescência , Oxirredução , FotoquímicaRESUMO
Recent studies demonstrated that the cationic, mitochondrial-specific dye Rhodamine-123 (Rh-123), is an efficient tumor photosensitizer for Argon laser treatment of human cancer cells both in vitro and in tumors grown as xenografts in athymic mice. To demonstrate the photodynamic mechanism of action of this reaction, the intracellular role of oxygen and temperature changes in treated cells have to be defined. In the current study, a large panel of human tumor cell lines of diverse histologic origin were tested for in vitro sensitivity to Rh-123 and the Argon laser (514.5 nm) in oxygen, deuterium oxide (D2O), and nitrogen (N2) environment. Tumor cells in suspension were first sensitized to Rh-123 (1 or 20 micrograms/ml for 1 hour), cooled on ice to 4 degrees C, and then exposed to the Argon laser (delta T = 14 +/- 1 degree C). Cell proliferation measured by [3H]-thymidine uptake 24 hours after sensitization with Rh-123 and laser treatment was significantly decreased in tumor cells kept in oxygen and D2O atmospheres. No decrease in DNA synthesis was seen in Rh-123 and laser treated cells kept in an N2 environment. Control tumor cells treated with Rh-123 or the Argon laser separately did not show any decreased [3H]-thymidine uptake in oxygen, D2O or N2 environment. These results provide evidence of a photodynamic process since Rh-123 sensitization and Argon laser activation occur at nonthermal levels of energy and are oxygen dependent. The high effectiveness of this technique of photodynamic therapy with the Argon laser, and low toxicity of Rh-123 could make its clinical use very attractive for the treatment of superficial malignancies.
Assuntos
Corantes Fluorescentes/uso terapêutico , Terapia a Laser , Fotoquimioterapia , Rodaminas/uso terapêutico , Xantenos/uso terapêutico , Deutério , Óxido de Deutério , Neoplasias de Cabeça e Pescoço/tratamento farmacológico , Humanos , Técnicas In Vitro , Nitrogênio , Oxigênio/fisiologia , Rodamina 123 , Células Tumorais Cultivadas , ÁguaRESUMO
Human eosinophils preferentially utilize bromide to generate a brominating agent, even at physiological halide concentrations, where chloride (140 mM) is over 1000-fold greater than bromide (20-100 microM). Under the same conditions, neutrophils use chloride to generate a chlorinating agent. The total amount of active halogen trapped by 1,3,5-trimethoxybenzene from eosinophils increases by over 2-fold as the added bromide concentration increases from 0 to 100 microM, with approximately 40 nmol of halogen trapped per million cells at the highest bromide level. At least 25-35% of the oxygen consumed by stimulated eosinophils is directed toward the generation of halogenating species. Since the relative halogenating behavior of eosinophil peroxidase and neutrophil myeloperoxidase in this bromide range is essentially identical to that of the cells, the specificity of eosinophils toward bromide is intrinsic to eosinophil peroxidase and not to any special cellular properties. These results suggest that human eosinophils use bromide in vivo and that a deficiency of bromide may influence their ability to produce halogenating agents.
Assuntos
Brometos/sangue , Cloretos/sangue , Eosinófilos/metabolismo , Neutrófilos/metabolismo , Soluções Tampão , Humanos , Técnicas In Vitro , Cinética , Consumo de Oxigênio , Peroxidase/sangue , Peroxidases/sangue , Floroglucinol/análogos & derivadosAssuntos
Catecóis , Oxigênio , Modelos Moleculares , Conformação Molecular , Oxirredução , Fotoquímica , Oxigênio SingleteRESUMO
The reactions of singlet oxygen (1O2) with cis and trans butenes-1,1,1-d3, at -80 degrees C in Freon-11, show a product isotope effect (kH/kD) of 1.38 and 1.25 respectively. Isomerization of the starting materials or formation of dioxetanes were not observed during the course of the photooxygenation. Together with the isotope effects on the reactions of tetramethylethylene-d6 isomers with singlet oxygen, these results require the reversible formation of a perepoxide or charge transfer intermediate.
Assuntos
Alcenos , Oxigênio , Deutério , Modelos Químicos , Oxirredução , EstereoisomerismoAssuntos
Fotoquímica , Fotoquimioterapia , Deutério , Cinética , Luz , Medições Luminescentes , Oxirredução , OxigênioRESUMO
On phagocytosing a microorganism, the neutrophil (polymorphonuclear leukocyte, PMN) consumes oxygen at a sharply elevated rate1. The oxygen is used to kill the microorganism, presumably being used to produce a potent oxidizing agent or agents. Candidates for these bactericidal agents are singlet oxygen, hydroxyl radical and chlorinating agents (that is, species containing 'active' Cl in a formal +1 oxidation state: HOCl, Cl2, N-chloroamides, and so on)1-5. We now report a semiquantitative assay for PMN-generated active chlorine based on its trapping with 1,3,5-trimethoxybenzene (TMB). Using this assay, we have found that at least 28% of the oxygen consumed by stimulated normal human PMNs is converted to active chlorinating agents.