Water-soluble Porphyrin Research Articles

Partial Unfolding of Tubulin Heterodimers Induced by Two-Photon Excitation of Bound meso-Tetrakis(sulfonatophenyl)porphyrin.

The water-soluble porphyrin meso-tetrakis(p-sulfonatophenyl)porphyrin (TSPP) can be noncovalently bound to tubulin and used as a photosensitizer, which upon irradiation triggers photochemical reactions that lead to conformational changes of the protein. These conformational changes in turn inhibit tubulin's primary function of polymerizing into microtubules. We explored the possibility of using two-photon excitation of the bound porphyrin to induce photosensitized protein unfolding. Although TSPP has a relatively low cross section (∼30 GM) our results did find that two-photon excitation of the ligand causes partial unfolding of the tubulin host and the inhibition of the in vitro formation of microtubules. Conversely, irradiating tubulin alone caused no such effects despite the large irradiance per pulse (97-190 GW/cm(2)). The conformational changes were characterized using spectroscopic studies and provide a promising protocol for the future application of non-native photosensitization of proteins.

Potential-Dependent Adsorption and Orientation of meso-Substituted Porphyrins at Liquid|Liquid Interfaces Studied by Polarization-Modulation Total Internal Reflection Fluorescence Spectroscopy

Potential-dependent adsorption behavior of meso-substituted water-soluble porphyrins at the polarized water|1,2-dichloroethane (DCE) interface was studied by polarization-modulation total internal reflection fluorescence (PM-TIRF) spectroscopy. In the PM-TIRF experiments, the fluorescence signal from the interfacial region was analyzed as a function of the periodic modulation of linear-polarizations (p and s) of the incident excitation beam. The potential-dependence of PM-TIRF responses for meso-substituted porphyrins, 5,10,15,20-tetrakis(N-methylpyridyl)porphyrin (H2TMPyP4+) and 5,10,15,20-tetrakis(4-sulfonatophenyl)porphyrin (H2TPPS4–), indicated that both free base porphyrins were adsorbed with relatively lying orientations at the polarized water|DCE interface. The average orientation angles (θ) were estimated as θ = 61 ± 1° for H2TMPyP4+ and θ = 65 ± 1° for H2TPPS4– with respect to the interface normal. The wavelength-dependence of polarization-modulated fluorescence signals (PM-TIRF spectrum), which ...

Multimerization of Solution-State Proteins by Water Soluble Porphyrins

Abstract: Interactions between charged porphyrins and complimentary or similarly charged proteins provide important models systems for studies of electron transfer processes, artificial photosynthesis, and control of protein-protein interactions. Typically, the experimental results are analyzed and discussed assuming that the proteins exist in a monodisperse state. Here, we explored interaction of four solution-state proteins (horse heart cytochrome c, hen egg-white lysozyme, 3-heme c-type cytochrome PpcA from Geobacter sulfurreducens, 2-heme cyt c4 from Pseudomonas stutzeri) with several cationic and anionic water-soluble derivatives of tetraphenylporphyrin. Combined small- and wide-angle X-ray scattering experiments revealed quick formation of mutlimers with a wide range of complex sizes. Thermodynamic interaction parameters and complex binding stoichiometries were established with isothermal calorimetry. Locations of porphyrin binding sites were determined with heteronuclear NMR and chemical modification shielding experiments. The obtained results demonstrate that multimerization of solution-state proteins by large water-soluble ligands appears to be a wide-spread phenomenon controlled by a delicate interplay of electrostatic and hydrophobic forces. Molecular level mapping of the binding sites allows us to build a theory explaining the size of the formed complexes and provides opportunities for targeted design and assembly of multi-subunit protein complexes.

Photo-Induced Charge and Energy in Complexes of C-Type Cytochromes with Water-Soluble Porphyrins

Photo-Induced Charge and Energy in Complexes of C-Type Cytochromes with Water-Soluble Porphyrins

Photophysical comparisons of PEGylated porphyrins, chlorins and bacteriochlorins in water

Synthesis of a bioconjugatable water-soluble (PEGylated) trans-AB-porphyrin enables photophysical comparisons (τS, kf, kic, kisc, Φf, Φic, Φisc) with analogous chlorins and bacteriochlorins in DMF and water.

A Novel Copper(II) Sensor Based on a Unique Water-soluble Porphyrin.

A unique water-soluble porphyrin [5,10,15,20-tetra(3-ethoxy-4-hydroxy-5-sulfonate)phenyl porphyrin, H2TEHPPS] was designed and synthesized, which could be used as a potential sensor for the determination of Cu(2+). Studies were performed in the solution. The concentration of H2TEHPPS was 5 μmol L(-1). The optical properties of H2TEHPPS were investigated based on the absorption spectra. The results show that the absorbance of H2TEHPPS at 417 nm is directly proportional to the concentration of Cu(2+) in the range of 0 - 2.5 μmol L(-1). H2TEHPPS can thus be used as a novel class of Cu(2+) sensors.

Electron transfer mediated decomposition of folic acid by photoexcited dimethoxophosphorus(V)porphyrin

Abstract A water soluble porphyrin, dimethoxophosphorus(V)tetraphenylporphyrin (MP(V)TPP) photosensitized folic acid decomposition in aqueous solution, resulting in the formation of a strongly fluorescent pteridine compound. The quantum yield of folic acid decomposition by photoexcited MP(V)TPP could be determined by a flurometry. A possible mechanism of the initial process of this photodecomposition is direct oxidation through an electron transfer from folic acid to the excited singlet state of MP(V)TPP. It is considered that this electron transfer proceeds as a diffusion-controlled reaction. The quantum yields of the electron transfer and the following process of folic acid decomposition could be determined. In deuterium oxide, folic acid decomposition through the photosensitized singlet oxygen generation by MP(V)TPP was also observed. A fluorometry based on the formation of a strong fluorescent compound through the oxidized decomposition of folic acid may be applied to evaluate the biomolecule damaging-activity of photosensitizers. In addition, a protocol of this assay is proposed. This simple photosensitized reaction in aqueous solution may be used for a first screening of a photosensitizer for phototherapy.

Resonance Raman and vibrational mode analysis used to predict ligand geometry for docking simulations of a water soluble porphyrin and tubulin

The ability to modify the conformation of a protein by controlled partial unfolding may have practical applications such as inhibiting its function or providing non-native photosensitive properties. A water-soluble porphyrin, meso-tetrakis (p-sulfonatophenyl) porphyrin (TSPP), non-covalently bound to tubulin can be used as a photosensitizer, which upon irradiation can lead to conformational changes of the protein. To fully understand the mechanism responsible for this partial unfolding and determine the amino acid residues and atoms involved, it is essential to find the most likely binding location and the configuration of the ligand and protein. Techniques typically used to analyze atomic position details, such as nuclear magnetic resonance and X-ray crystallography, require large concentrations, which are incompatible with the dilute conditions required in experiments for photoinduced mechanisms. Instead, we develop an atomistic description of the TSPP–tubulin complex using vibrational mode analysis from density functional theory calculations correlated to resonance Raman spectra of the porphyrin paired with docking simulations. Changes in the Raman peaks of the porphyrin molecule correlate with changes in its structural vibrational modes when bound to tubulin. The data allow us to construct the relative geometry of the porphyrin when bound to protein, which are then used with docking simulations to find the most likely configuration of the TSPP–tubulin complex.

Development of PDT/PET Theranostics: Synthesis and Biological Evaluation of an (18)F-Radiolabeled Water-Soluble Porphyrin.

Synthesis of the first water-soluble porphyrin radiolabeled with fluorine-18 is described: a new molecular theranostic agent which integrates the therapeutic selectivity of photodynamic therapy (PDT) with the imaging efficacy of positron emission tomography (PET). Generation of the theranostic was carried out through the conjugation of a cationic water-soluble porphyrin bearing an azide functionality to a fluorine-18 radiolabeled prosthetic bearing an alkyne functionality through click conjugation, with excellent yields obtained in both cold and hot synthesis. Biological evaluation of the synthesized structures shows the first example of an (18)F-radiolabeled porphyrin retaining photocytotoxicity following radiolabeling and demonstrable conjugate uptake and potential application as a radiotracer in vivo. The promising results gained from biological evaluation demonstrate the potential of this structure as a clinically relevant theranostic agent, offering exciting possibilities for the simultaneous imaging and photodynamic treatment of tumors.

A porphyrin-PEG polymer with rapid renal clearance

Tetracarboxylic porphyrins and polyethylene glycol (PEG) diamines were crosslinked in conditions that gave rise to a water-soluble porphyrin polyamide. Using PEG linkers 2 kDa or larger prevented fluorescence self-quenching. This networked porphyrin mesh was retained during dialysis with membranes with a 100 kDa pore size, yet passed through the membrane when centrifugal filtration was applied. Following intravenous administration, the porphyrin mesh, but not the free porphyrin, was rapidly cleared via renal excretion. The process could be monitored by fluorescence analysis of collected urine, with minimal background due to the large Stokes shift of the porphyrin (230 nm separating excitation and emission peaks). In a rhabdomyolysis mouse model of renal failure, porphyrin mesh urinary clearance was significantly impaired. This led to slower accumulation in the bladder, which could be visualized non-invasively via fluorescence imaging. Without further modification, the porphyrin mesh was chelated with 64Cu for dynamic whole body positron emission tomography imaging of renal clearance. Together, these data show that small porphyrin-PEG polymers can serve as effective multimodal markers of renal function.

Phototoxic Activity and DNA Interactions of Water-Soluble Porphyrins and Their Rhenium(I) Conjugates.

In the search for alternative photosensitizers for use in photodynamic therapy (PDT), herein we describe two new water-soluble porphyrins, a neutral fourfold-symmetric compound and a +3-charged tris-methylpyridinium derivative, in which either four or one [1,4,7]-triazacyclononane (TACN) units are connected to the porphyrin macrocycle through a hydrophilic linker; we also report their corresponding tetracationic Re(I) conjugates. The in vitro (photo)toxic effects of the compounds toward the human cell lines HeLa (cervical cancer), H460M2 (non-small-cell lung carcinoma), and HBL-100 (non-tumorigenic epithelial cells) are reported. Three of the compounds are not cytotoxic in the dark up to 100 μm, and the fourfold-symmetric couple revealed very good phototoxic indexes (PIs). The intracellular localization of all derivatives was studied in HeLa cells by confocal fluorescence microscopy. Although low nuclear localization was observed for some of them, it still prompted us to investigate their capacity to bind both quadruplex and duplex DNA; we observed significant selectivity in the tris-methylpyridinium derivatives for G-quadruplex interactions.

Influence of H2TOEtPyP4 porphyrin on the stability and conductivity of bilayer lipid membranes.

Many water-soluble cationic porphyrins are known to be prospective chemotherapeutics and photosensitizers for cancer treatment and diagnosis. The physicochemical properties of porphyrins, in particular their interactions with membranes, are important determining factors of their biological activity. The influence of cationic meso-tetra-[4-N-(2'-hydroxyethyl) pyridyl] porphyrin (H2TOEtPyP) on the stability and conductivity of bilayer lipid membranes (BLMs) was studied. H2TOEtPyP4 porphyrin was shown to decrease the stability of BLMs made of a mixture of DOPS and DPPE (1:1) in an electric field because of a reduction of line tension of spontaneously formed pore edges in the BLM. The presence of cationic porphyrin was found to reduce BLM surface tension. This effect was enhanced with increasing porphyrin concentration. H2TOEtPyP4 increased the probability of spontaneous pore formation. Further investigating the cyclic current-voltage characteristics of BLMs allowed determining the electrical capacity and conductivity of BLMs in the presence of H2TOEtPyP4 porphyrin. It was shown that in the presence of cationic porphyrin the electrical capacity as well as conductivity of the BLM increases.

Sn(IV) Metalloporphyrin/Co(III) Complex: An All-Abundant-Element System for the Photocatalytic Production of H2 in Aqueous Solution.

A new, molecular system for the light-driven production of hydrogen in aqueous solution was developed by combining a water-soluble tin porphyrin ([Sn(IV)Cl2TPPC], A) acting as photosensitizer with a cobalt-based proton-reduction catalyst ([Co(III)Cl(dmgH)2(py)], C). Under visible light illumination and with triethanolamine (TEOA) as electron source, the system evolves H2 for hours and is clearly catalytic in both dye and catalyst. A detailed analysis of the relevant redox potentials in combination with time-resolved spectroscopy resulted in the development of a Z-scheme type model for the flow of electrons in this system. Key intermediates of the proposed mechanism for the pathway leading to H2 are the porphyrin dye's highly oxidizing singlet excited state (1)A* (E ∼ +1.3 V vs NHE), its strongly reducing isobacteriochlorin analogue (E ∼ +0.95 V), and the Co(I) form of C (E ∼ -0.8 V), acting as catalyst for H2 formation. Among other results, the suggested reaction sequence is supported by the detection of a shortened excited-state lifetime for singlet (1)A* (τ ∼ 1.75 ns) in the presence of TEOA and the ultraviolet-visible detection of the Sn(IV) isobacteriochlorin intermediate at λ = 610 nm. Thus, a molecular, conceptually biomimetic, and precious-metal-free reaction chain was found which photocatalytically generates H2 in a 100% aqueous system from an electron donor with a high oxidation potential (E(TEOA) ∼ +1.1 V). On the other hand, at identical conditions, this photoreaction chain yields H2 markedly slower than a system using the photosensitizer [Re(I)(CO)3(bpy) (py)](+), probably due to the much longer excited-state lifetime (τ ∼ 120 ns) of the rhenium dye and better electron-transfer rates caused by its simple single-electron photoreduction chemistry.

Effect of Pluronic F127 on the photosensitizing properties of dimegine in the presence of nanoparticles

It is shown that the activity of the water-soluble porphyrin photosensitizer dimegine (DMG) in the oxidation reaction of tryptophan, a test reaction for photodynamic therapy (PDT), can be enhanced by introducing silver, gold, and hydroxyapatite nanoparticles (NP) into the reaction medium if an amphiphilic polymer (AP) with the properties of a SAS is introduced into this mixture. It is concluded that the effect of enhancing the photosensitizing activity of dimegine is due to the formation of nanoparticle-Pluronic-porphyrin triple systems in which Pluronic (PL) plays the role of a bridge, forming complex and adsorption bonds with porphyrin and nanoparticles.

Synthesis and photophysical properties of the photoactivatable cationic porphyrin 5-(4-N-dodecylpyridyl)-10,15,20-tri(4-N-methylpyridyl)-21H,23H-porphyrin tetraiodide for anti-malaria PDT.

This article describes a new synthetic method for obtaining three water soluble porphyrins. The more sophisticated porphyrin [5-(4-N-dodecylpyridyl)-10,15,20-tri(4-N-methylpyridyl)-21H,23H-porphyrin tetraiodide], also named C12 porphyrin, was obtained through a three step methodology. The improvements, compared to syntheses described in the literature, mostly concern the purification procedures. The photophysical properties of the three porphyrins are described and the C12 porphyrin presents a very good (1)O2 yield compared to its chemical intermediates. This porphyrin seems to be a very promising candidate for PDT applications.

Effect of chemical structure of bipyridinium salts as electron carrier on the visible-light induced conversion of CO2 to formic acid with the system consisting of water-soluble zinc porphyrin and formate dehydrogenase

Effect of chemical structures of some 2,2′-bipyridinium salts (BP2+) as the electron carrier molecules on the visible-light induced conversion of CO2 to formic acid with the system consisting of water-soluble zinc tetraphneylporphyrin tetrasulfonate (ZnTPPS) and formate dehydrogenase (FDH) in the presence of triethanolamine (TEOA) as an electron donor molecule was investigated. Irradiation of a CO2 saturated solution containing TEOA, ZnTPPS, BP2+ and FDH with visible light resulted in production of formic acid. By using 1,1′-ethylene-2,2′-bipyridinium dibromide (DB2+) as an electron carrier molecule, the effective formic acid production was observed compared with the other 2,2′-bipyridinium salt derivatives.

Electrocatalytic Water Oxidation by a Water-Soluble Nickel Porphyrin Complex at Neutral pH with Low Overpotential.

The water-soluble cationic nickel(II) complex of meso-tetrakis(4-N-methylpyridyl)porphyrin (1) can electrocatalyze water oxidation to O2 in neutral aqueous solution (pH 7.0) with the onset of the catalytic wave appearing at ∼1.0 V (vs NHE). The homogeneous catalysis with 1 was verified. Catalyst 1 exhibited water oxidation activity in a pH range 2.0-8.0 and had a strict linear dependence of catalytic current on its concentration. After 10 h of constant potential electrolysis at 1.32 V (vs NHE), a negligible difference of the solution was observed by UV-vis. In addition, inspection of the working electrode by electrochemistry, scanning electron microscope (SEM), and energy dispersive X-ray spectroscopy (EDX) showed no sign of deposition of NiOx films. These results strongly argued that 1 is a real molecular electrocatalyst for water oxidation. The turnover frequency (TOF) for this process was 0.67 s(-1) at 20 °C. On the basis of results from the kinetic isotope effect (KIE) and inhibition experiments, electrochemical studies in various buffer solutions with different anions and pHs, and DFT calculations, a catalytic cycle of 1 for water oxidation via a formally Ni(IV) species was proposed.

Asymmetric Oxidation and Carbene Transfer Reactions Catalyzed By Metalloporphyrins in Water

The development of environmentally benign reactions is an important goal in synthetic organic chemistry and chemical engineering. However, catalytic enantioselective reactions using transition-metal complexes in protic or aqueous solvents are within limited ranges. The current applications of asymmetric catalysis using metalloporphyrins will be reported (1). Distinct methodologies have been developed for carbene transfer and oxidation reactions such as the use of water-soluble catalysts (2). Comparison between hydrophobic and hydrophilic solvents will be discussed from an asymmetric point of view. Catalytic enantioselective oxidations using transition-metal complexes are limited when the oxidant is hydrogen peroxide. The two main difficulties of using hydrogen peroxide in the presence of transition metal complexes are the homolytic cleavage generating OH radicals and the catalase reaction with formation of dioxygen. Asymmetric sulfoxidation, epoxidation of alkenes and hydroxylation catalyzed by iron and manganese porphyrins will be described (3). Thus the asymmetric epoxidation of styrene derivatives by H2O2 (or UHP) to give optically active epoxides (ee up to 81%) and hydroxylation of alkanes to give optically active secondary alcohols (ee up to 78%) were carried out in methanol and water using chiral water-soluble iron porphyrins as catalysts. (1) Recent Advances in Catalysis by Water-Soluble Metalloporphyrins G. Simonneaux, P. Le Maux, S. Chevance and H. Srour. Handbook of Porphyrin Science, 2012, Vol. 21, 377-410 (2) Enantioselective Manganese-Porphyrin-Catalyzed Epoxidation and C-H hydroxylation with Hydrogen Peroxide in Water/Methanol Solutions H. Srour, P. Le Maux and G. Simonneaux. Inorg. Chem. 2012, 51, 5850-5856. (3) Metal-catalyzed asymmetric sulfoxidation, epoxidation and hydroxylation by hydrogen peroxide Hassan Srour, Paul Le Maux, Soizic Chevance, Gérard Simonneaux. Coord. Chem. Rev., 257 (2013) 3030– 3050

New Generation of Hybrid Materials Based on Hierarchical Assembly of Porphyrin on Supramolecular Scaffolds or Nanostructures

Porphyrins multifunctional properties make these macrocycles intringuing building blocks to build up complex supramolecular assemblies via covalent or non-covalent approaches, either in solution or on inorganic surfaces. In the perspective of development of hybrids organic-inorganic materials, mastering of non-covalent strategies to transfer water soluble porphyrin based assemblies on nanostructured surfaces paves the way to a cost effective and environmentally friendly research that combine the unique properties of nanomaterials with the synthetic versatility of organic chemistry. Noteworthy, the control of spontaneous self-assembly is not trivial, as it is a hierarchically driven process that can proceed through thermodynamic or kinetic pathways. However, in water, complex porphyrin based supramolecular assemblies having a pre-determined and controlled stechiometry, chirality and composition have been successfully synthetized. Transfer of these assemblies (i.e. chiral porphyrin chiral aggregates, porphyrin-calixarenes, porphyrin-DNA) on inorganic surfaces opens the way to development of new generation of chiral surfaces and sensors, where porphyrins played an important role as spectroscopic probes, capable to overcome sensitivity limitations. Chirality induction and memory is, accordingly, another important requirement that need to be kept during this migration from solution to the solid state. The robustness of non-covalent approaches to achieve a straightforward, controllable and environmentally friendly functionalization of inorganic nanostructures strongly hinges on the competition between retention at the solid state rather than in solution and depends on several factors. Among them, polyelectrolytes secondary structure strongly influences the outcome of spontaneous deposition of porphyrin assemblies on solid surfaces often overriding the mere system electrostatics.

Synthesis of Porphyrin-Based Nanostructures By Ionic Self-Assembly: Exploring Novel Tectons

Self-assembly of porphyrins is a powerful method for the production of highly functionalized nanomaterials. Porphyrin-based nanomaterials can be self-assembled using a number of procedures including ionic self-assembly (ISA), re-precipitation and coordination polymerization.1 ISA has proven to be a particularly flexible synthetic method and can be used to couple oppositely-charged porphyrin ions (tectons) in a range of environments such as at liquid-liquid interfaces,2 on templates,3 or spontaneously in water.4,5 The resulting materials have been investigated as catalysts for the reduction of O2 to H2O,2 as photosensitizers for the reduction of H2O to H2,6 as adsorbed pigments for functionalizing carbon nanotubes,7 as H2 storage materials,8 and as photoconductors.5 Despite these considerable advances, little is known about the ISA of structurally modified porphyrin tectons that may be useful for producing nanomaterials with specific properties. It is well established that extension of the π-system or nonplanar deformation of the macrocycle significantly alter many properties of porphyrins, including their optical and electronic behaviour. In this talk, we describe recent investigations of the self-assembly of tecton 1 (M = Ni)9 which contains both of these structural modifications, with the standard tecton TPPS 2(M = Sn, Fe, Mn, Cu or 4H) (see Graphic). Porphyrins 1 and 2 are shown to react to produce nanoscale materials together with porous micron-sized wafers, although the reactions are significantly different than those previously reported for ISA of standard water-soluble porphyrins based on the TPP framework. Of particular note are the reactions of 1 with the diprotonated form of 2 (M = 4H)whichcan yield precipitates with stoichiometries lower than or much higher than those expected for ISA. Reasons for the formation of these unusual products are discussed, together with the potential applications of this chemistry to the formation of ternary systems containing 1, a metal complex of 2, and the diprotonated form of 2. [1] For a recent review see: C. J. Medforth, Z. Wang, K. E. Martin, Y. Song, J. L. Jacobsen, J. A. Shelnutt, Chem. Commun.7261 (2009). [2] A. J. Olaya, D. Schaming, P.-F. Brevet, H. Nagatani, T. Zimmermann, J. Vanicek, H.-J. Xu, C. P. Gros, J.-M. Barbe, H. H. Girault, J. Am. Chem. Soc. 134,498 (2012). [3] R. Lauceri, G. F. Fasciglione, A. D'Urso, S. Marini, R. Purrello, M. Coletta, J. Am. Chem. Soc. 130, 10476 (2008). [4] Z. Wang, C. J. Medforth, J. A. Shelnutt, J. Am. Chem. Soc. 126,15954 (2004). [5] K. E. Martin, Z. Wang, T. Busani, R. M. Garcia, Z. Chen, Y. Jiang, Y. Song, J. L. Jacobsen, T. T. Vu, N. E. Schore, B. S. Swartzentruber, C. J. Medforth, J. A. Shelnutt, J. Am. Chem. Soc. 132, 8194 (2010). [6] Y. Tian, K. E. Martin, J. Y.-T. Shelnutt, L. Evans, T. Busani, J. E. Miller, C. J. Medforth, J. A. Shelnutt, Chem. Commun. 47, 6069 (2011). [7] A. Brewer, M. Lacey, J. R. Owen, I. Nandhakumar, E. Stulz, J. Porphyrins Phthalocyanines 15, 257 (2011). [8] M. T. Oztek, M. D. Hampton, D. K. Slattery, S. Loucks, Int. J. Hydrogen Energy 36, 6705 (2011). [9] L. Jiang, R. A. Zaenglein, J. T. Engle, C. Mittal, C. S. Hartley, C. J. Ziegler, H. Wang, Chem. Commun. 48, 6927 (2012). Figure 1