RESUMEN
Resveratrol (3,5,4'-trihydroxystilbene, RSV) is a natural stilbene synthetized as trans-isomer in plants exposed to oxidative stress. In order to understand the mechanism involved during photosensitized degradation of trans-resveratrol, steady-state and time-resolved experiments were performed and compared with quantum-chemical calculations using density functional theory (DFT). Pterin (Ptr), a well-known photosensitizer, under UV-A radiation induces the oxidation of several biomolecules mainly through electron-transfer mechanisms. On the one hand, it was observed that trans-RSV participates in an energy-transfer pathway with Ptr triplet excited state (3Ptr*) forming 3trans-RSV*, which dissipates the energy by isomerization to cis-RSV. On the other hand, RSV neutral radical (trans-RSV(-H)â¢) was detected in laser flash photolysis experiments, evidencing an electron-transfer mechanism. The electron-transfer from 3Ptr* to trans-RSV is a barely feasible reaction, however, more favorable is the formation of trans-RSV(-H)⢠in a reaction between trans-RSV and Ptr radical cation (Ptrâ¢+), which is produced during irradiation. The combination of experimental and theoretical approaches evidences the capability of trans-RSV to undergo energy-transfer (feasible by DFT calculations) and/or one-electron transfer pathways with 3Ptr*. These findings reveal the mechanisms involved in the interaction of trans-RSV and pterin excited states and provide information on the antioxidant action of resveratrol during photosensitized oxidation of biomolecules.
Asunto(s)
Antioxidantes , Electrones , Resveratrol , Isomerismo , Antioxidantes/química , Pterinas/farmacologíaRESUMEN
Ricin is a ribosome-inactivating protein (RIP type 2) consisting of two subunits, ricin toxin A (RTA) and ricin toxin B (RTB). Because of its cytotoxicity, ricin has worried world authorities for its potential use as a chemical weapon; therefore, its inhibition is of great biotechnological interest. RTA is the target for inhibitor synthesis, and pterin derivatives are promising candidates to inhibit it. In this study, we used a combination of the molecular docking approach and fast steered molecular dynamics (SMD) to assess the correlation between nonequilibrium work, ⟨ W⟩, and the IC50 for six RTA inhibitors. The results showed that molecular docking is a powerful tool to predict good bioactive poses of RTA inhibitors, and ⟨ W⟩ presented a strong correlation with IC50 ( R2 = 0.961). Such a profile ranked the RTA inhibitors better than the molecular docking approach. Therefore, the combination of docking and fast SMD simulation was shown to be a promising tool to distinguish RTA-active inhibitors from inactive ones and could be used as postdocking filtering approach.
Asunto(s)
Antitoxinas/química , Antitoxinas/farmacología , Pterinas/química , Pterinas/farmacología , Ricina/antagonistas & inhibidores , Ricina/metabolismo , Sustancias para la Guerra Química/química , Sustancias para la Guerra Química/metabolismo , Humanos , Ligandos , Simulación del Acoplamiento Molecular , Ricina/química , Ricinus/químicaRESUMEN
A new series of decyl chain [-(CH2)9CH3] pterin conjugates have been investigated by photochemical and photophysical methods, and with theoretical solubility calculations. To synthesize the pterins, a nucleophilic substitution (SN2) reaction was used for the regioselective coupling of the alkyl chain to the O site over the N3 site. However, the O-alkylated pterin converts to N3-alkylated pterin under basic conditions, pointing to a kinetic product in the former and a thermodynamic product in the latter. Two additional adducts were also obtained from an N-amine condensation of DMF solvent molecule as byproducts. In comparison to the natural product pterin, the alkyl chain pterins possess reduced fluorescence quantum yields (ΦF) and increased singlet oxygen quantum yields (ΦΔ). It is shown that the DMF-condensed pterins were more photostable compared to the N3- and O-alkylated pterins bearing a free amine group. The alkyl chain pterins efficiently intercalate in large unilamellar vesicles, which is a good indicator of their potential use as photosensitizers in biomembranes. Our study serves as a starting point where the synthesis can be expanded to produce a wider series of lipophilic, photooxidatively active pterins.
Asunto(s)
Fármacos Fotosensibilizantes/farmacología , Pterinas/farmacología , Oxígeno Singlete/química , Alquilación , Fluorescencia , Concentración de Iones de Hidrógeno , Interacciones Hidrofóbicas e Hidrofílicas , Cinética , Fosfolípidos/química , Fármacos Fotosensibilizantes/química , Pterinas/química , Solubilidad , Solventes/químicaRESUMEN
UV-A radiation (320-400nm), recognized as a class I carcinogen, induces damage to the DNA molecule and its components through different mechanisms. Pterin derivatives are involved in various biological functions, including enzymatic processes, and it has been demonstrated that oxidized pterins may act as photosensitizers. In particular, they accumulate in the skin of patients suffering from vitiligo, a chronic depigmentation disorder. We have investigated the ability of pterin (Ptr), the parent compound of oxidized pterins, to photosensitize the degradation of the pyrimidine nucleotide thymidine 5'-monophosphate (dTMP) in aqueous solutions under UV-A irradiation. Although thymine is less reactive than purine nucleobases, our results showed that Ptr is able to photoinduce the degradation of dTMP and that the process is initiated by an electron transfer from the nucleotide to the triplet excited state of Ptr. In the presence of molecular oxygen, the photochemical process leads to the oxidation of dTMP, whereas Ptr is not consumed. In the absence of oxygen, both compounds are consumed to yield a product in which the pterin moiety is covalently linked to the thymine. This compound retains some of the spectroscopic properties of Ptr, such as absorbance in the UV-A region and fluorescence properties.
Asunto(s)
Oxidación-Reducción/efectos de los fármacos , Fármacos Fotosensibilizantes/farmacología , Pterinas/farmacología , Timidina Monofosfato/química , Transporte de Electrón/efectos de los fármacos , Humanos , Oxígeno/química , Nucleótidos de Purina/química , Timidina Monofosfato/efectos de la radiación , Rayos UltravioletaRESUMEN
Microbial related contamination is of major concern and can cause substantial economic losses. Photodynamic inactivation (PDI) has emerged as a suitable approach to inhibit microorganism proliferation. In this work, PDI induced by 6-carboxypterin (Cap), a biocompatible photosensitizer (PS), was analyzed. The growth inhibition of Staphylococcus aureus exposed to artificial UV-A radiation and sunlight in the presence of Cap was investigated. After UV-A irradiation, 50 µM Cap was able to decrease by three orders (with respect to the initial value) the number of S. aureus cells in early biofilms. However, this concentration was 500 times higher than that needed for eradicating planktonic cells. Importantly, under solar exposure, 100 µM Cap was able to suppress sessile bacterial growth. Thus, this strategy is able to exert a bactericidal effect on sessile bacteria and to eradicate planktonic cells by exposing the Cap-containing sample to sunlight.
Asunto(s)
Biopelículas/efectos de los fármacos , Fotoquimioterapia/métodos , Fármacos Fotosensibilizantes/farmacología , Plancton/efectos de los fármacos , Pterinas/farmacología , Staphylococcus aureus/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Staphylococcus aureus/fisiologíaRESUMEN
Pterins are normal components of cells and they have been previously identified as good photosensitizers under UV-A irradiation, inducing DNA damage and oxidation of nucleotides. In this work, we have investigated the ability of pterin (Ptr), the parent compound of oxidized pterins, to photosensitize the oxidation of another class of biomolecules, amino acids, using tryptophan (Trp) as a model compound. Irradiation of Ptr in the UV-A spectral range (350 nm) in aerated aqueous solutions containing Trp led to the consumption of the latter, whereas the Ptr concentration remained unchanged. Concomitantly, hydrogen peroxide (H2O2) was produced. Although Ptr is a singlet oxygen ((1)O2) sensitizer, the degradation of Trp was inhibited in O2-saturated solutions, indicating that a (1)O2-mediated process (type II oxidation) was not an important pathway leading to Trp oxidation. By combining different analytical techniques, we could establish that a type I photooxidation was the prevailing mechanism, initiated by an electron transfer from the Trp molecule to the Ptr triplet excited state, yielding the corresponding radical ions (Trp(·+)/Trp(-H)· and Ptr(·-)). The Trp reaction products that could be identified by UPLC-mass spectrometry are in agreement with this conclusion.
Asunto(s)
Oxidación-Reducción , Fármacos Fotosensibilizantes/metabolismo , Pterinas/metabolismo , Triptófano/metabolismo , Espectroscopía de Resonancia por Spin del Electrón , Peróxido de Hidrógeno/metabolismo , Peróxido de Hidrógeno/efectos de la radiación , Nucleótidos/metabolismo , Fármacos Fotosensibilizantes/farmacología , Pterinas/farmacología , Oxígeno Singlete/metabolismo , Triptófano/efectos de los fármacos , Triptófano/efectos de la radiación , Rayos UltravioletaRESUMEN
Pterins, heterocyclic compounds widespread in biological systems, are able to photoinduce oxidation of DNA and its components. In the present study, we have investigated the photosensitizing properties of pterin (Ptr), the parent compound of oxidized pterins, using bovine serum albumin (BSA) as target. Aqueous solutions of BSA were exposed to UV-A irradiation (350nm) in the presence of Ptr, under various experimental conditions. The photosensitized processes were followed by UV/vis spectrophotometry, an enzymatic method for H2O2 determination and electrophoresis (SDS-PAGE). We present data that demonstrate unequivocally that BSA is damaged by Ptr. Although association between Ptr and the protein was evidenced by steady-state and time-resolved fluorescence measurements, the photosensitized damage takes place via a purely dynamic mechanism, which involves an electron transfer from BSA to the triplet excited state of Ptr, formed after UV-A excitation.
Asunto(s)
Fármacos Fotosensibilizantes/farmacología , Pterinas/farmacología , Albúmina Sérica Bovina/metabolismo , Animales , Bovinos , Fármacos Fotosensibilizantes/metabolismo , Pterinas/metabolismoRESUMEN
Tyrosinase catalyzes in mammals the first and rate-limiting step in the biosynthesis of the melanin, the main pigment of the skin. Pterins, heterocyclic compounds able to photoinduce oxidation of DNA and its components, accumulate in the skin of patients suffering from vitiligo, a chronic depigmentation disorder in which the protection against UV radiation fails due to the lack of melanin. Aqueous solutions of tyrosinase were exposed to UV-A irradiation (350 nm) in the presence of pterin, the parent compound of oxidized pterins, under different experimental conditions. The enzyme activity in the irradiated solutions was determined by spectrophotometry and HPLC. In this work, we present data that demonstrate unequivocally that the enzyme is photoinactivated by pterin. The mechanism of the photosensitized process involves an electron transfer from tyrosinase to the triplet excited state of pterin, formed after UV-A excitation of pterin. The biological implications of the results are discussed.
Asunto(s)
Monofenol Monooxigenasa/antagonistas & inhibidores , Monofenol Monooxigenasa/efectos de la radiación , Pterinas/farmacología , Fármacos Sensibilizantes a Radiaciones/farmacología , Agaricales/enzimología , Dihidroxifenilalanina/química , Monofenol Monooxigenasa/química , Oxidación-Reducción , Procesos Fotoquímicos , Rayos UltravioletaRESUMEN
Pterins, heterocyclic compounds widespread in biological systems, participate in relevant biological processes and are able to act as photosensitizers. In the present study, we ascertained that 2-aminopteridin-4(3H)-one, abbreviated as Ptr, is readily incorporated into and/or onto cervical cancer cells (HeLa) and that these cells die upon UV-A irradiation of Ptr. Cell death was assessed using two tests: (1) the Rhodamine 123 fluorescence assay for mitochondrial viability and (2) the Trypan Blue assay for membrane integrity. The data suggest that, for Ptr-dependent photoinitiated cell death, events related to mitochondrial failure precede those associated with the failure of the cell membrane.